WO2016198756A1

WO2016198756A1 - Cosmetic, nutraceutical, veterinary and pharmaceutical compositions containing a hazelnut pericarp extract

Info

Publication number: WO2016198756A1
Application number: PCT/FR2016/000094
Authority: WO
Inventors: Philippe Bernard; Franck HIMBERT
Original assignee: Greenpharma
Priority date: 2015-06-12
Filing date: 2016-06-08
Publication date: 2016-12-15
Also published as: FR3037244B1; FR3037244A1

Abstract

The invention relates to cosmetic, nutraceutical, veterinary and pharmaceutical uses of a hazelnut pericarp extract as an agent for the treatment of skin, mucus or skin appendages.

Description

COSMETIC COMPOSITIONS,

NUTRACEUTIQUES, VETERINARIES AND PHARMACEUTICALS CONTAINING AN EXTRACT FROM

PERICARP OF HAZELNUT

The present invention relates to the use of extracts of a plant that is widespread, particularly in Europe, in cosmetic, pharmaceutical, nutraceutical or veterinary compositions. More specifically, it relates to new applications of hazelnut pericarp for treating human or animal skin.

The Hazel or Coudrier (Corylus avellana L.) is a shrub 3 to 8 meters high and belonging to the family Betulaceae. It is a plant of woods, hedges and gardens that gives a popular edible fruit, hazelnut. He has a soft wood. He is sometimes called Avelinier. The shrub forms a tuft of 10 to 12 branches up to 3 to 4 m high, multi-stemmed (composed of several thin trunks). Its bark is brown and can come off in thin strips depending on the variety. Its deciduous heart-shaped leaves are toothed with a pointed top. The yellow, yellow flowers form pendent ears or 6 cm kittens, and the highly condensed female flowers form erect ears.

Hazelnut is an achene endowed with a woody pericarp called "shell" and containing a single seed (the almond) which occupies the entire internal cavity of the pericarp. This nut, more or less ovoid in shape, can reach 3 cm long and 2 in diameter; it is protected before complete maturity by a tubular envelope, the involucre (bract), of foliaceous aspect and divided into irregular lobes at its end, it is more or less enveloping according to the varieties.

The pericarp is a waste of the agro-food industry and is found in large quantities. Most of the time, he underwent a roasting step.

Hazelnut pericarp has been studied for its antioxidant properties in view of the tannins it contains (Alasalvar et al., Antioxidant activity of haselnut skin phenolics, J. Agric., Food Chem (2009) 57, 4645). Various scientific publications also describe the tannins it contains (Del Rio et al., Phenolic composition of haselnut skin, J. Agric Food Chem (2011) 59, 9935) or sugars (Montella et al, Identification and characterization of water alkali soluble oligosaccharides from haselnut skin (C. avellana L.) Food Chemistry 140 (2013) 717-725.

The present invention demonstrates new applications of extracts of hazelnut pericarp for at least partially depigmenting normal skin or at least partially removing unsightly brown spots related to age or repeated exposure to the sun for example. But, in addition, extracts of hazelnut pericarp significantly improve the barrier function of the epidermis to fight against external aggression and thus plays a role of skin protector. These extracts of hazelnut pericarp also play a favorable role in the anti-aging compositions. Finally, these extracts of pericarp have a lipolytic role.

According to the invention it has been discovered that hazelnut pericarp extracts make it possible to reduce the expression of tyrosinase, the key enzyme of pigmentation in normal human melanocytes. This result makes it possible to consider depigmenting actions on naturally pigmented skin or on sun-induced pigment spots, aging or any other external aggressions. The present invention therefore aims at providing new cosmetic or pharmaceutical, nutraceutical or veterinary compositions capable of reducing the natural or induced pigmentation of the skin of a living animal and in particular of a human subject.

According to the invention it has also been discovered that hazelnut pericarp extracts make it possible to stimulate the expression of various essential protein markers in the formation of the stratum corneum or stratum corneum, including the key enzyme TGK transglutaminase.

The stratum corneum is the most superficial part of the skin and represents the culmination of the keratinocyte differentiation process. During this differentiation, basement-level keratinocytes undergo a series of metabolic and structural changes throughout their migration to the skin surface. The last stage is their transformation into corneocytes whose accumulation in a cohesive structure constitutes the stratum corneum which provides a barrier function. Throughout the process of differentiation, several proteins intervene. One of them, TGK, of the glutaminase family, catalyzes the establishment of covalent type B (g-glutamyl) -lysine peptide bonds between various protein precursors that form the stratum corneum and confers on it this ability to protect skin against external aggressions and limit the diffusion of water from the inside. Transglutaminase forms generally insoluble protein polymers. These biological polymers are essential for the body to create barriers and stable structures. According to the invention we have finally demonstrated a lipolytic effect of hazelnut pericarp extracts by acting on the release of non-esterified fatty acids in human adipocytes.

The present invention aims precisely at providing new cosmetic, pharmaceutical, nutraceutical or veterinary compositions capable of stimulating the factors involved in differentiation, called pro-differentiating factors such as TGK, and thus making it possible to improve the conditions of the skin, of the dander and mucous membranes, healthy or injured.

According to the invention, said compositions comprise, as active principle, at least one hazelnut pericarp extract.

"Hazelnut pericarp extract" means an extract or a mixture of plant extracts of the genus Corylus sp of the family Betulaceae. It is more particularly an extract or a mixture of extracts of cells of Corylus sp. This cellular material can be obtained by in vitro culture or in vivo. By in vitro culture is meant all the techniques known to those skilled in the art, which allow, artificially, to obtain a plant or part of a plant. Thus, the extract may be an extract or a mixture of organ extracts (root, stem, leaf, bark), or even of organ cells, of at least one plant of the genus Corylus sp of the Betulaceae family. or an undifferentiated cell extract from at least one such plant.

The applications according to the invention extend to the treatment of the skin, superficial body growths and mucous membranes. By dander, include nails and the hair system, especially the hair. Mucous membranes are the covering tissues of the anatomical cavities, which are connected to the skin by natural orifices, such as the mouth, stomach, intestine, as well as those of natural external cavities such as the nostrils or the ears.

Thus, the subject of the present invention is a use of a hazelnut pericarp extract for depigmenting and / or protecting the skin, superficial body growths and mucous membranes of external physical, chemical or biological agents and / or for improving and / or strengthening moisturizing the skin and / or strengthening the integuments and mucous membranes and / or having a lipolytic action.

It also relates to a cosmetic treatment method for combating the dryness of a skin or for the cosmetic treatment of dry skin, said method comprising a step according to which at least one hazelnut pericarp extract is applied to the skin, to restore flexibility and / or elasticity.

The invention also relates to a method of cosmetic anti-aging treatment of the skin or cosmetic treatment of an aged skin, comprising a step according to which at least one hazelnut pericarp extract is applied to the skin. This treatment can delay the phenomena of pigmentation and / or aging of the skin, but also to repair an aged skin by restoring flexibility and elasticity.

In any cosmetic application of the invention, a hazelnut pericarp extract may be combined with another or other skin-producing agents, such as calcium, vitamin D and their derivatives or at least one other known complementary depigmenting agent. of the skilled person.

According to another aspect, the invention relates to pharmaceutical, nutraceutical or veterinary applications of a hazelnut pericarp extract, as presented hereinafter.

One of the objects according to the invention is therefore a pharmaceutical, nutraceutical or veterinary composition comprising an extract of hazelnut pericarp, to restore the barrier function of the epidermis of an injured skin. This state can result from the action of a physical, chemical or biological external agent and / or a disorder or a disease, such as for example psoriasis or atopic dermatosis.

Another object according to the invention is a pharmaceutical, nutraceutical or veterinary composition comprising a hazelnut pericarp extract, for improving and / or reinforcing the barrier function of a mucosa or for restoring the barrier function of an injured mucosa. For example, it may be the intestinal mucosa. The lesion of the mucosa may result from a disorder or a disease; in the case of the intestinal mucosa, the lesion may be caused by irritable bowel syndrome.

In the therapeutic aim of the invention, the hazelnut pericarp extract may be administered topically; it can also be administered orally.

According to one aspect of the invention, the extract is administered after the inflammatory process is completed, in particular repair treatment by reconstruction of the stratum corneum.

In a composition of the invention, the hazelnut pericarp extract may also be combined with one or more anti-inflammatory and / or immunosuppressive agents used in the treatment of cutaneous inflammation, such as cyclosporins and derivatives thereof, anti-cytokine biotherapies and corticosteroids.

A hazelnut pericarp extract according to the invention can be obtained by any extraction or purification method known to those skilled in the art. In particular, mention may be made of solid-liquid extraction processes in alcoholic (in particular ethanolic) and aqueous media, as well as in mediums using solvents such as ketones, esters, ethers, polyols, chlorinated solvents and mixtures of at least two of the abovementioned solvents, such as hydroalcoholic media.

Advantageously, an extract is obtained by extraction of hazelnut pericarp, milled or not, in aqueous hydroalcoholic or alcoholic medium, in particular with an aqueous solution of ethanol. The concentration of ethanol can vary from 10 to 90% (v / v).

Alternative methods to solvents can also be considered as supercritical C0 ₂ or microwaves ...

These extracts can be used as such, in liquid or powder form, unpurified or purified.

If the extract is powdered, its drying can be conducted by any technique well known to those skilled in the art. The extract can be spray dried, evaporated or lyophilized. The powder thus obtained may be encapsulated in liposomes or other vectors and supports for a better homogeneity of the composition and a better diffusion of the active principle, in particular on the skin.

The extract is preferably obtained from the pericarp of hazelnuts crushed or not and roasted or not.

It has been observed that the extracts obtained by hydroalcoholic maceration of the hazelnut pericarp exhibit a high biological activity, regardless of the percentage of ethanol in the extraction solvent. Similar results are observed in water or ethanol.

The extracts of hazelnut pericarp may be combined in the compositions according to the invention with substances increasing skin protection. By way of example but in a nonlimiting manner, mention may be made of associations with mucopolysaccharides, vitamins, ceramides, antiradical substances or U.V. filters.

Similarly, the reparative activity of the extracts according to the invention is particularly advantageous when they are associated with substances having a healing effect such as proteins, hyaluronic acid, amino acids, and / or with anti-inflammatory substances, anti-aging, after-sun or with active anti-acne and anti-dermatoses.

The protective activity of the extracts of the invention with respect to free radicals and UV is also very interesting in the hair field, especially in the case of combination with substances facilitating the good condition of the scalp and that hair, such as minerals, vitamins, ceramides, protein extracts, mucopolysaccharides, flower or fruit acids. The compositions of the invention are particularly suitable for topical application for preventing and / or treating numerous skin disorders.

The subject of the invention is therefore the dermo-cosmetic use of a composition containing at least one hazelnut pericarp extract, as a depigmenting agent for a naturally pigmented or partially pigmented skin, in order to homogenize the complexion, or to reduce brown spots related to age or external agents such as the sun. The invention therefore also relates to the dermocosmetic use of a hazelnut pericarp extract, as a repairing and / or protective agent for the skin and the hair system, such as the hair, in particular for combating external aggression, such as that those related to pollution, sun, oxidative stress, aging and skin diseases leading to dysfunction of the homeostasis of the epidermis or hair. Therefore, a direct application of the compositions of the invention relates to the fight against aging and cell death induced by the U.V.

For a cosmetic use, the compositions of the invention may be in the form of creams, gels, lotions, milks, O / W and W / O emulsions, solutions, ointments, sprays, body oils, hair lotions, shampoos, lotions. aftershave, soaps, lip sticks, sticks and pencils for makeup. The compositions may comprise any excipients necessary for their formulation. Thus, when in the form of gel, they contain suitable excipients such as cellulose esters or other gelling agents such as Carbopol ^®, guar gum or the like.

These cosmetic compositions may also take the form of a lotion or solution in which the extracts and / or molecules are in encapsulated form, for example in microspheres. These microspheres may for example consist of fat, agar-agar and water. The active agents can also be incorporated into liposome, glycosphere-type vectors in chylomicrons, macro-, micro-, nano-particles as well as macro-, micro- and nano-capsules and can also be adsorbed onto polymers. powdery organic, talcs, bentonites and other mineral supports. These emulsions have good stability and can be stored for the time necessary for use at temperatures between 0 and 50 ° C without sedimentation of constituents or phase separation.

The cosmetic compositions of the invention comprise of the order of

0.01 to 5% by weight, preferably between 0.1 and 2.5%, of pericarp extract of hazelnut when they are in the form of powder and of the order of 0.01 to 25% by weight, preferably between 0.5 and 10%, when they are in encapsulated form. The percentages given above are expressed by weight of dry extract of hazelnut pericarp relative to the weight of the final composition.

For the preparation of these compositions, the hazelnut pericarp extracts are mixed with the excipients generally used in cosmetics.

The cosmetic compositions of the invention may also contain additives or adjuvants which are customary in cosmetology, for example antibacterial agents or perfumes, but also extraction and / or synthetic lipids, gelling and viscosity polymers, surfactants and the like. active and emulsifying agents, hydro or liposoluble active ingredients, plant extracts, tissue extracts, marine extracts and synthetic actives.

The cosmetic or nutraceutical compositions according to the invention can be used to exert a lipolytic action by topical application or orally.

The cosmetic or dermocosmetic use of extracts includes all body and skin care including sun, protective and tanning products, lipolytics, anti-aging products, anti-seborrhoeic products, tonics, products that improve the skin. the appearance of the skin including acne treatment, skin rash, scalp treatment and hair loss.

The cosmetic compositions of the present invention may also comprise other complementary active ingredients chosen for their action, for example for sun protection, the anti-wrinkle effect, the antiradical and antioxidant activity, the anti-irritant activity, the cellular nutrition, cellular respiration, hydration and cell regeneration, anti-seborrhoeic treatments, lipolysis such as caffeine and other active ingredients having an effect on skin tone, protection of the hair, eyelashes and nails

The cosmetic compositions of the present invention are preferably used daily by applying them one or more times per day. The cosmetic compositions of the present invention are very well tolerated, they exhibit no phototoxicity and their application to the skin for prolonged periods of time does not imply any systemic effect.

The invention also relates to the use of hazelnut pericarp extracts for the preparation of dermocosmetic compositions having an anti-inflammatory and dermoprotective activity. These compositions are useful for preventing and / or treating in particular dermatological diseases related to seborrheic, acneic and / or inflammatory activities. These dermocosmetic compositions are in liquid form, powder, paste or emulsion, alone or in combination with other substances. They comprise of the order of 0.01 to 25% by weight of hazelnut pericarp extract. The percentages are expressed by weight of dry extract of hazelnut pericarp relative to the weight of the final composition. For the preparation of these dermocosmetic compositions, extracts of hazelnut pericarp are mixed with excipients.

Consequently, in general, the invention relates in particular to the use of a hazelnut pericarp extract, for the preparation of a pharmaceutical, veterinary, nutraceutical, dermatological or cosmetic composition intended to prevent and / or treat the pathologies and disorders resulting:

■ natural or premature aging of the skin or hair, promoting the detoxification and purification of the skin.

^■ dermatoses, such as psoriasis, atopic dermatitis ...,

^■ inflammatory conditions of the cutaneous or hair systems, inducing itching, irritation, chronic redness, irritation and persistent itching and functional disorders of capillary fragility or nails

^■ skin pigmentation,

^■ disorders related to adipocytes, such as excess fat storage, dimpling, loss of the oval of the face.

A pharmaceutical composition according to the invention comprises, in addition to at least one active agent as defined above, a pharmaceutically acceptable vector or excipient.

An oral nutraceutical composition according to the invention comprises, in addition to at least one active agent as defined above, a vector or excipient acceptable in terms of food supplements. This nutraceutical composition can target any organ such as the skin or an internal organ such as the intestine.

The present invention is now illustrated, without limitation, by the following Examples 1 to 5. Example 1 illustrates processes for preparing extracts of hazelnut pericarp according to the invention.

Examples 2, 3 and 4 illustrate the biological properties claimed according to the invention. Example 2 shows the depigmenting effects of certain extracts according to the invention. Example 3 illustrates the properties of the extracts of Example 2, which were evaluated by the ability of these extracts to stimulate the expression of key markers of epidermal differentiation. The selected markers, all involved in the arrangement of keratin filaments and in the formation of the horny layer envelope, are transglutaminase K (TGK), filaggrin and cytokeratin (KRT10). TGK catalyzes the establishment of s (g-glutamyl) -lysine peptide bonds between various protein precursors. Filaggrin and KRT10 are poorly expressed in primary keratinocyte monolayers and are considered markers of terminal differentiation of the epidermis. These results testify to the ability of extracts to act on aged or injured skin. These properties can be extended to other organs of the human or animal body. Example 4 demonstrates the lipolytic effect of the extracts. Finally, Example 5 is a case of cosmetic formula.

Example 1: Obtaining extracts of hazelnut pericarp

A series of extractions was carried out with the following solvents

• hexane;

• ethyl acetate ;

• ethanol / water: 90/10;

• ethanol / water: 60/40;

• ethanol / water: 30/70;

• water.

The extractions were carried out at room temperature (except for water at 50 ° C.), after a preliminary grinding of the plant material. The plant / solvent ratio is 1/10. The maceration time (with stirring) is 24 hours.

For each extract, one part was treated on activated carbon with a coal / plant ratio of 1/10, the other was kept raw.

The extraction yields are collated in Table 1 below:

It is first noted that the hexane and ethyl acetate extracts have a yield close to 10%, which seems to indicate the presence of a high proportion of fat. This is confirmed by the oily nature of these extracts. It should be noted that the aqueous extract and 30% ethanol were very difficult to filter. It is possible that these difficulties led to losses that decrease

10 artificial way yields. The yield seems optimum for 60% ethanol: 16% crude and 13% treated on charcoal. When the amount of ethanol is reduced, the reduction in yield caused by the coal treatment increases sharply. This is consistent with the high levels of phenolic compounds described in the next section. It is found that the treatment on

Coal is, apart from oils, rather inefficient in diminishing the color of the extracts.

The phytochemical analyzes of these extracts are as follows:

Total phenolic compounds and tannins were determined by the Folin-Ciocalteu method in chlorogenic acid equivalent. The results are summarized in Table 3 below:

We observe that :

The higher the extraction solvent is rich in ethanol, the more the extract is rich in phenolic compounds;

Among the phenolic compounds, tannins are a minority;

• from 60% ethanol, the charcoal treatment makes it possible to enrich the extract with phenolic compounds and tannins;

• the 90% and 60% ethanol extracts, treated on carbon contain respectively 98% and 95% of phenolic compounds; of which respectively 25% and

10 24% tannins.

Example 2 Evaluation of the Cytotoxicity and the Depigmenting Activity of the Extracts

15

We assessed the tyrosinase inhibitory activity for the untreated ethanol 30% (Ex3MF1173) and ethanol 90% (ExIMF1173) extracts.

Cytotoxicity

We have a 1 ^st time assessed the cytotoxicity of extracts of melanocytes.

• Cell type: NHEM in culture medium.

• Incubation time: 72 hours.

25 · Evaluation parameters reduction of MTT and morphological observations under the microscope. At the end of the treatment, the cells were incubated in the presence of MTT (tetrazolium salt) whose transformation into blue crystals of formazan is proportional to the activity of succinate dehydrogenase (mitochondrial enzyme). After dissociation of cells and solubilization of formazan by addition

From DMSO, the optical density (OD), representative of the number of living cells and their metabolic activity, was measured with a 540 nm microplate reader (VERSAmax, Molecular Devices). From the results presented in Table 4 below it is concluded that the maximum non-cytotoxic dose is 3 μg / ml for ExlMF1173 and 10 μ§ / ιηί for Ex3MF1173.

Tyrosinase activity

The melanocytes were inoculated and cultured in culture medium for 24 hours. The medium was then replaced with medium containing or not (control) the compounds or the reference (lipoic acid at 5 ° C.).

and the cells were incubated for 72 hours.

All the experimental conditions were realized in n = 3.

At the end of the incubation, the culture medium was removed and the cells washed with PBS solution. For each condition, the tyrosinase enzyme was extracted into PBS-Triton X-100 solution, and then incubated with the substrate (2 mM L-DOPA) for 1 hour at 37 ° C.

The enzymatic activity of the various cell extracts was evaluated by measuring the optical density (OD) at 540 nm (Versamax microplate reader, Molecular Devices) and using a standard range of fungal tyrosinase (0.39 to 400 U / ml).

The activity of tyrosinase was evaluated after 72 hours of NHEM culture in the presence of the reference or compounds under test. In this type of protocol, the measurement of the enzymatic activity makes it possible to account for the amount of tyrosinase present in the melanocytes. However, it can not be ruled out that the compounds, incorporated in the cells or covalently bound to the enzyme, can directly inhibit the enzymatic activity of tyrosinase.

After 72 hours of culture, tyrosinase activity was detected in the NHEMs of the control condition. The treatment of NHEM with lipoic acid, tested at 5 μg / ml, induced a very strong inhibition of tyrosinase activity (90% inhibition). These results were expected and validated the trial.

Extract EX3MF1173, tested at 10 μg / ml, inhibited the tyrosinase activity (25% inhibition). At the lowest concentrations tested, this compound had no effect. The aqueous effect also showed depigmenting activity.

Extract EX1MF1173, tested at 3 concentrations, did not modulate the activity of tyrosinase. However, it should be noted that the extract ExlMF1173, because of its greater cytotoxicity, was tested at a concentration three times lower than Ex3MF1173. At this same concentration Ex3MF1173 also has no effect. It is therefore possible that the only difference between ExlMF1173 and Ex3MF1173 lies only in their non-cytotoxic maximum levels.

The results are shown in Table 5 below:

Two extracts of hazelnut pericarp emerge from these first trials.

¹ is the crude ethanol extract 30% (Ex3MF1173) having an inhibitory effect of tyrosinase (25%) significant. It contains about 60% of phenolic compounds (chlorogenic acid equivalent), about 5% of which are tannins (chlorogenic acid equivalent). The extraction yield is about 7%.

In order to evaluate the color impact in a cosmetic formula, the two extracts will be diluted to 10% in maltodextrin and formulated in a cream at 1%. Formulas and actives will be accelerated to assess their stability in color and phenol content. Example 3 Evaluation in primocultures of normal human epidermal keratinocytes (NHEK)

1) Materials and methods

cells

Normal human epidermal keratinocytes (NHEK); reference

BlOalternatives K341, used in the third pass.

Culture conditions: 37 ° C, 5% C0 ₂

Culture medium: KeratinocyteSFM supplemented with epidermal growth factor (EGF) 0.25ng / ml, pituitary extract (PE) 25μg / ml and gentamycin

Test medium: KeratinocyteSFM supplemented with gentamycin

25μg / ml.

Extracts tested

In the initial step of selecting an extract for further testing, and after evaluation of prior cytotoxicity, extracts EX8MF1113 and EX9MF1113 are tested at the following concentrations:

The above step led to these two extracts being retained at non-cytotoxic concentrations of 10 μg / ml; dose used for the following experiments.

Immunolabeling in situ

Keratinocytes were seeded and cultured at confluence in plates

96 wells then the medium was replaced with test medium containing or not (control) the tested extract or reference (CaCl ₂ at 1.5 mM) and the cells were incubated for 72 hours. All experimental conditions were performed in n = 3, for each marker.

After incubation, the test medium is removed and the cells are rinsed, fixed and permeabilized. Cells are labeled with primary antibodies against the proteins of interest (TGK, filaggrin and KRT10) according to Mcheik JN et al. Foreskin-isolated keratinocytes provide successful extemporaneous autologous pediatric skin grafts. J Tissue Eng Regen Med. 2013 Mar 14. These antibodies were revealed by a secondary antibody coupled to a fluorochrome (GAMAlexa 488). In parallel, the nuclei of the cells are stained with Hoechst 33258 (bisbenzimide). Image acquisition is performed with a high resolution imaging system, INCell Analyzer ™ 1000 (GE Healthcare). For each well, 5 entries of scanned images are made. The labels are quantified by measuring the fluorescence intensity of the proteins relative to the number of nuclei identified by the Hoechst (digital data integration by the software Developer Toolbox 1.5, GE Healthcare).

Extraction screening was performed only on the TGK parameter (table

6).

Transcript analysis, RT-qPCR

The keratinocytes were inoculated and grown to confluency in 24-well plates and then the medium was replaced with test medium containing or not (control) the test compound or reference (CaCl ₂ at 1.5 mM) and the cells were incubated for 48 hours. All experimental conditions were performed in n = 3, for each marker.

After incubation, the test medium was removed and the cells were rinsed and frozen at -80 ° C.

The expression of the markers was evaluated by RT-qPCR on the messenger RNAs (mRNAs) extracted from the cell mats of each treatment (the replicates were pooled before the extraction of the RNA). The steps of RNA extraction, reverse transcription, primers and PCR conditions have been previously described, according to Mcheik JN et al. Foreskin-isolated keratinocytes provide successful extemporaneous autologous pediatric skin grafts. J. Tissue Eng Regen Med. 2013 Mar 14; Boniface et al. IL-22 inhibitory epidermal differentiation and proinflammatory induces gene expression and migration of human keratinocytes. J Immunol. 2005 Mar 15; 174 (6): 3695-702; Boniface et al. Oncostatin M secreted by skin infiltrating T lymphocytes is a potent keratinocyte activator involved in skin inflammation. J Immunol. 2007 Apr. 178 (7): 4615-22.

2) Results

screening

The results are shown in Table 6 below, which illustrates the level of expression of TGK via fluorescent labeling of NHEKs, the fluorescence intensity of NHEK being proportional to TGK expression. Table 6

These results show that the EX9MF1113 extract induces a dose-dependent expression of TGK in NHEK cultures. The activity is greater than that of the reference calcium chloride, at a concentration of 1 μg / ml; the maximum effect, which is 4 times greater than that of CaCl _2, is observed at the non-cytotoxic concentration of 10 μg / ml Influence of the extract EX9MF1113 on the expression of TKG, filaggrin and KRT10

The same methodology used to evaluate the effect of an extract on TGK expression was applied to the markaggrine and KRT10 markers.

Table 8 provides an assessment of the level of expression of the above three markers via fluorescent labeling of NHEKs.

In this experiment, one finds the inducing effect of TGK and one observes a very strong overexpression of protein KRT10. Unlike calcium, which focuses the effect on a few cells by transforming them into very large differentiated cells (corneocytes), EX9MF1113 induces a very strong overexpression of KRT10 in many cells but does not significantly increase cell size. The mechanism of action of calcium and EX9MF1113 are therefore different and could be additional or synergistic. This is confirmed by the analysis of filaggrin, which increases (more moderately) following treatment with EX9MF1113, but without inducing the characteristic production of filaggrin granules in calcium-responsive cells. This example opens up perspectives towards the oral or topical positioning of the extract to redensify the hair, the eyelashes, the nails but also to restore the barrier function, the hydration, and to improve in a general way the manifestations related to age.

Example 4: Evaluation of the effects of hazelnut pericarp extracts on the release of non-esterified fatty acids by human adipocytes.

The purpose of this test is to measure the hydrolysis of triglycerides in human hypodermic adipocytes. The reference compound is caffeine. The weather incubation time is two hours. The results are shown in Table 9 below:

In this model, the extracts behave as the positive control. One can imagine a positioning topically or orally of such an extract to reduce skin imperfections related to adipocytes such as cellulite or to restore the oval face in anti-aging care.

Example 5: Dermo-cosmetic formulation of an extract of hazelnut pericarp.

Non-limitingly, we present below an example of formula in emulsion form in Table 10 below.

Claims

1. Cosmetic, pharmaceutical or nutraceutical composition intended at least partially, depigmenting and / or protecting the skin, the mucous membranes and the integuments of a living animal, characterized in that it contains at least one active agent containing at least one extract of hazelnut pericarp.

2. Pharmaceutical composition according to claim 1, characterized in that it protects the skin and the mucous membranes by improving and / or reinforcing their barrier function.

3. Pharmaceutical composition according to claim 2, characterized in that it is used to restore the barrier function of a skin damaged by a physical, chemical or biological external agent and / or by a disease such as dermatitis, atopic dermatitis or psoriasis.

4. Pharmaceutical or nutraceutical composition according to claim 1, characterized in that it is used to improve and / or strengthen the barrier function of a mucosa by topical application or orally.

5. Cosmetic or nutraceutical composition according to claim 1, characterized in that it is used to exert a lipolytic action by topical application or orally.

6. Pharmaceutical or nutraceutical composition according to claim 4, characterized in that it is used orally to improve or enhance the barrier function of an injured intestinal mucosa.

7. Pharmaceutical or nutraceutical composition according to claim 1, characterized in that it is used Orally or topically to redensify hair, eyelashes and / or nails.

8. Pharmaceutical or nutraceutical composition according to claim 1, characterized in that it is used

5 to restore the barrier function of the skin, hydration of the skin and to improve the manifestations related to age.

9. Pharmaceutical or nutraceutical composition according to claim 1, characterized in that it is used

10 to reduce cutaneous imperfections related to adipocytes, such as cellulite and / or to restore the oval of the face as anti-aging care.

10. Pharmaceutical composition according to one of claims 2 to 5, characterized in that the extract of

The hazelnut pericarp it contains is associated with at least one other skin-producing agent, especially calcium, vitamin D and their derivatives.

11. Pharmaceutical composition according to one of claims 2 to 6, characterized in that it comprises

At least one anti-inflammatory and / or immunosuppressive agent used in. the treatment of cutaneous inflammation, in particular cyclosporins and their derivatives, anti-cytokine biotherapies and corticosteroids.

^'25

12. Cosmetic use of a composition according to claim 1, characterized in that it improves and / or enhances the hydration of the skin, by topical or oral administration.

13. Cosmetic use of a composition according to claim 1, by topical or oral administration, for the depigmentation of the skin of a human subject by decreasing the expression of tyrosinase in normal human melanocytes.

14. Cosmetic use according to claim 9, characterized in that one treats a naturally pigmented skin or induced pigmentary spots.

15. Cosmetic use according to claim 8, in which a dry or aged skin is treated by applying the composition to the skin in order to increase its suppleness and / or elasticity.

16. Cosmetic use of a composition according to claim 1, characterized in that the barrier function of the skin is protected or reinforced against an external physical, chemical or biological agent by topical or oral administration.

17. Process for obtaining the hazelnut pericarp extract of claim 1, characterized in that a solid / liquid extraction of said hazelnut pericarp is carried out in a solvent medium comprising at least one solvent taken from the group formed. by water, alcohols, ketones, ethers, polyols, chlorinated solvents and supercritical C0 ₂ .

18. The method of claim 16, characterized in that the solid / liquid extract is produced in a water / ethanol mixture in a proportion ranging from 10% to 90% by volume.