Use of a tetrapeptide with the formula N-Palmitoyl-Lys-Thr-Phe-Lys in the treatment of human atopic dermatitis

Abstract

The invention relates to a tetrapeptide of the formula N-Palmitoyl-Lys-Thr-Phe-Lys, for its use in the prevention and / or treatment of human atopic dermatitis (AD), and to a composition comprising at least said tetrapeptide and at least one physiologically acceptable excipient, for its use in the prevention and / or treatment of human atopic dermatitis (AD). Figure to be published with the abstract: 1

Description

Title of the invention: Use of a tetrapeptide of formula N-Palmitoyl-Lys-Thr-Phe-Lys in the treatment of human atopic dermatitis

[0001] The present invention is in the field of atopic dermatitis and in particular the prevention and / or treatment of human atopic dermatitis.

[0002] More specifically, the present invention relates to a tetrapeptide of formula N-Palmitoyl-Lys-Thr-Phe-Lys, for its use in the prevention and / or treatment of human atopic dermatitis (AD), as well as to a composition comprising at least said tetrapeptide and at least one physiologically acceptable excipient, for its use in the prevention and / or treatment of human atopic dermatitis (AD).

[0003] According to the Scientific Council of the French Society of Allergology, atopic dermatitis, also called atopic eczema, is a chronic inflammatory skin disease. It develops primarily in infants and children, but can persist or even appear in adolescents and adults. It is characterized by dry skin (lipid deficiency combined with water deficiency) associated with eczema-like lesions (redness and itching, blisters, oozing, and crusting) that occur in flare-ups. In babies, the lesions are most often found on the chubby areas of the face and limbs, on the scalp, and on the buttocks. Later, they are found in skin folds (neck, behind the knees, elbow creases, eyelids), on the hands, and around the mouth. The majority of atopic dermatitis cases disappear during childhood (50% before age 5), but 10 to 15% of cases persist into adulthood.

[0004] The most frequent complications of atopic dermatitis are colonization of skin lesions by Staphylococcus aureus, leading to dysbiosis of the skin microbiome, or by the herpes virus. Some dermatitis may be associated with growth delays and ophthalmological complications (frequent keratoconjunctivitis, cataracts, retinal detachment).

[0005] Like many chronic diseases, atopic dermatitis can have a significant psychological impact, causing sleep disorders, irritability or even depressive syndrome.

[0006] The term "atopy" refers to a hereditary predisposition to developing an allergy, which can manifest as dermatitis, but also as asthma or allergic rhinitis. Family studies have shown that atopic dermatitis is indeed associated with genetic predisposition factors: between 50 and 70% of Affected individuals have a first-degree relative who is also affected. If both parents are affected, the risk of the child developing allergic eczema reaches 80%.

[0007] It is likely that several genes contribute to the emergence of the pathology. Recent studies have notably highlighted abnormalities affecting the gene encoding filaggrin and other molecules necessary for the formation of the stratum corneum and the resistance of the skin barrier.

[0008] The increase in the prevalence of atopic dermatitis observed over the last forty years indicates that environmental changes play a role in the phenomenon, as genetic causes cannot explain such a rapid increase. Many factors can contribute to this sudden rise: excessive hygiene and loss of immune system stimulation at an early stage of development, excessive skin washing, very isolated and poorly ventilated housing conducive to dust mites, increased presence of pets, early evolution and diversification of dietary habits, exposure to tobacco and urban industrial pollution, later pregnancies, and children more often cared for in group settings... However, it is difficult to estimate the relative weight of these factors, which are complex in nature and often act in combination.

[0009] Unlike contact eczema, atopic dermatitis does not occur when the child touches a specific substance. A genetic predisposition is likely responsible for an alteration of the skin barrier in patients, causing dryness and increased sensitivity to irritants.

[0010] This alteration of the skin's structure will also allow environmental allergens (pollen, dust, soaps, etc.) to penetrate the epidermis and stimulate the immune system. The latter will overreact to what it perceives as an attack, triggering eczema: redness, itching, inflammation, and oozing.

[0011] The treatment of atopic dermatitis is based primarily on topical care. A long-term treatment aims to combat dry skin and restore the skin's barrier function. This involves the daily application of emollients (moisturizing products).

[0012] During a flare-up, treatment aims to reduce inflammation and itching, to relieve the patient and prevent superinfection. The application of a topical corticosteroid, an immunomodulator, an antiseptic solution, possibly with antihistamine tablets as an adjunct, helps the patient get through the attack.

[0013] Atopic dermatitis therefore requires drug treatment during flare-ups, but also the use of maintenance or comfort products between flare-ups. It is these maintenance and comfort products that are the subject of the present invention.

[0014] Emollient and lipid-replenishing products for atopic-prone skin already exist on the cosmetics market. Among such products, one can notably mention thermal water-based products sold under the trade names Avène, Uriage or La Roche-Posay, as well as oat-based products such as those sold under the trade name A-Derma®.

[0015] Furthermore, patent FR3118416B1 describes a cosmetic composition useful for the treatment of atopic dermatitis comprising a synergistic combination of at least one amino acid selected from lysine, histidine, arginine, ornithine, and hydroxyproline, and a glycosylceramide. This combination restores the functionality of tight junctions in pre-atopic and atopic skin.

[0016] However, there is still a need for topical formulations for atopic-prone and atopic skin, usable to prevent and / or treat human atopic dermatitis.

[0017] The invention described below makes it possible to meet this need.

[0018] The Inventors have demonstrated that a tetrapeptide with a specific formula, as defined below, reduces the overexpression of thymic stromal lymphopoietin (TSLP) and can thus be advantageously used in the prevention and / or treatment of atopic dermatitis. TSLP is a cytokine produced by keratinocytes and mast cells. Its production is greatly increased during atopic dermatitis and promotes the initiation of a Th2-type inflammatory response by dendritic cells (Carmi-Levy I et al., Clin. Rev. Allerg. Immunol., 2011; 41: 245-253).TSLP thus plays an important role in the atopic march (from atopic dermatitis to asthma) since its levels are correlated with skin sensitization and asthma severity in mice, whereas this sensitization and its cutaneous and bronchial consequences are greatly reduced in mice whose gene encoding TSLP is inactivated (Leyva-Castillo JM et al., J. Invest. Dermatol., 2013; 133: 154-163).

[0019] The invention therefore has as its first object a tetrapeptide of formula N-Pahnitoyl-Lys-Thr-Phe-Lys (also called tetrapeptide Pal-KTFK), for its use as an active ingredient for the prevention and / or treatment of human atopic dermatitis.

[0020] The tetrapeptide with the formula N-Palmitoyl-Lys-Thr-Phe-Lys therefore comprises a substitution by a palmitoyl chain at the N-terminus and no substitution at the C-terminus, and, in that order, the following amino acids: lysine (Lys), threonine (Thr), phenylalanine (Phe), and lysine (Lys). The tetrapeptide Pal-KTFK is a known compound, notably used in cosmetics in the form of oil-wax-surfactant-water microemulsions. It is, in particular, present in the product sold under the Crystalide® is a product sold under the trade name Crystalide® by Sederma. According to the International Nomenclature of Cosmetic Ingredients (INCI), the tetrapeptide in Crystalide® promotes the synthesis of the transparent α-crystalline protein found in the lens of the eye (approximately 50% of its content), but also distributed throughout many tissues, including the skin. α-crystalline is a chaperone protein that contributes to the skin's transparency and radiance. It can therefore be used to improve complexion and in formulations designed to combat the effects of aging.

[0021] Its role in reducing TSLP overexpression had, however, never been mentioned in the literature, let alone highlighted.

[0022] According to the invention, the tetrapeptide Pal-KTFK can be implemented in a topical, cosmetic or dermatological composition.

[0023] Thus, the present invention has as its second object a topical composition comprising at least one physiologically acceptable excipient and, as an active ingredient, at least one tetrapeptide of formula N-Pahnitoyl-Lys-Thr-Phe-Lys, for its use for the prevention and / or treatment of human atopic dermatitis. DEFINITIONS

[0024] According to the invention, a topical composition is understood to be a cosmetic or dermatological composition intended to be applied to the skin, including that of the scalp, and mucous membranes.

[0025] For the purposes of the present invention, the prevention of atopic dermatitis includes the treatment of skin with an atopic (or pre-atopic) tendency.

[0026] Atopic (or pre-atopic) skin is an extremely fragile skin type. It is characterized by an impaired skin barrier that promotes the penetration of allergens, resulting in dry skin that is more vulnerable to external aggressors and a predisposition to allergies. Atopic (or pre-atopic) skin can easily develop atopic dermatitis.

[0027] According to the invention, a physiologically acceptable excipient is an excipient suitable for external topical administration, in particular an excipient acceptable from a dermatological and cosmetic point of view. These excipients suitable for the formulation of a topical composition are well known to those skilled in the art. DETAILED DESCRIPTION

[0028] According to a particular embodiment of the invention, the topical composition may comprise at most about 10 ppm of said tetrapeptide.

[0029] According to a preferred embodiment, the content of said tetrapeptide within said composition is approximately 2 to 8 ppm, and even more preferably approximately 4 to 6 ppm relative to the total weight of said topical composition.

[0030] According to a particular embodiment, said tetrapeptide is used in the form of an emulsion, in particular an oil-in-water emulsion comprising one or more fatty acid triglycerides. As an example of such a tetrapeptide formulated as an emulsion, mention may be made of the product sold under the name Crystalide® by the company Sederma.

[0031] According to a particular embodiment of the invention, the topical composition further comprises calcium ions.

[0032] The presence of calcium ions in the topical composition usable according to the present invention is advantageous because these calcium ions contribute to the proper functioning of the skin's barrier function. Calcium ions serve as essential signaling molecules that regulate numerous cellular functions in keratinocytes, including proliferation and differentiation. Since keratinocytes require different calcium concentrations for various functions (low calcium for proliferation, high calcium for differentiation), an epidermal calcium gradient is formed. In the lower layers of the epidermis (basal layer and stratum spinosum), the calcium gradient is at its lowest but rises, reaching its peak in the granular layer, and then declines again towards the outermost layers of the stratum corneum, where keratinocytes reach their final stage of differentiation.This keratinocyte differentiation is mandatory for proper skin barrier function. In addition to driving keratinocyte differentiation, the calcium gradient plays a role in keratinocyte migration and wound healing. Taken together, the calcium gradient and signaling are essential for a healthy skin barrier and barrier homeostasis. Collapse of the calcium gradient disrupts skin homeostasis. In skin prone to atopic dermatitis, calcium signaling is dysregulated and the calcium gradient is flattened. The loss of the epidermal calcium gradient may partly result from fewer calcium-sensitive receptors. This leads to significant dysregulation of proteins that constitute the skin barrier, including loricrin and filaggrin, thus resulting in reduced barrier function.

[0033] According to a particularly advantageous embodiment of the invention, the calcium ions are encapsulated in a supramolecular structure comprising one or more concentric lipid bilayers enclosing aqueous compartments containing said calcium ions (vectorized calcium). These supramolecular structures are generally in the form of liposomes. or cones. In this type of structure, the lipid bilayers are formed by amphiphilic lipids, generally phospholipids, and calcium ions are present in the aqueous compartments between the lipid bilayers. An example of calcium delivery is the product sold under the trade name EpiCalsome® by Mibelle Biochemistry.

[0034] When calcium ions are present in the topical composition usable according to the invention, their content is preferably from 13 to 32 ppm approximately, and even more preferably from 19 to 25 ppm approximately, relative to the total weight of said composition.

[0035] In addition to the tetrapeptide as defined above, the topical composition usable according to the invention may further contain one or more additional active ingredients whose presence may enhance and / or complement the advantageous properties of the tetrapeptide as defined above in the prevention and / or treatment of atopic dermatitis. Such additional active ingredients may include, in particular, anti-pruritic and / or anti-neurogenic inflammation agents, agents that rebalance the cutaneous immune system, and agents that act on the dysbiosis of the cutaneous microbiome, including agents that inhibit the action of S. aureus or reduce its action.

[0036] As non-limiting examples of anti-pruritic agents, thermal waters such as the thermal waters of Avène, Uriage and La Roche Posay can be cited; saccharide gums obtained by fermentation of sorbitol, such as the biosaccharide gum-2 sold by the company Solabia, and certain plant extracts such as an extract of Boswellia serrata, Acmella oleracea, Aloe vera, oats (Avena sativa), burdock (Arclium iappa), calendula or marigold (Calendula), German chamomile (Mairiearia recutita), fumitory (Fumaria officinalis), witch hazel (Hamamelis virginiana), lavender (Lavandula officinalis or angustifolia or vera), mallow (Malva sylvestris), lemon balm (Melissa officinalis), common or stinging nettle (Urtica dioica or Urtica urens), wild pansy (Viola tricolor), horsetail fields (Equisetum arvenses), sarsaparilla (Smilax aspera), tea tree (Melaleuca altemifolia), white birch (Betula alba),red sage (Salvia miltiorrhiza), heart pea (Cardiospermum halicacabum), etc., corn or oat starch hydrolysates, and oily active ingredients such as peroxidized corn oil (Zea mays) and bisabolol.

[0037] Examples of cutaneous immune system rebalancing agents include, but are not limited to, unsubstituted [3-glucans such as the product sold under the trade name Plerasan Re-Balance® by ROELMI HPC, as well as [3-glucans substituted with carboxymethyl groups such as the product sold under the trade name CM-Glucan Forte (Magnesium Carboxymethyl Beta-Glucan) by the company Mibelle Biochemistry.

[0038] Examples of active ingredients acting on skin microbiome dysbiosis include, but are not limited to, alpha-glucan oligosaccharides, Yacon tuber extract (Polymnia sonchifolid), Lactobacillus, rhamnose, glucose, glucuronic acid, glycolipids such as flocculosin, which is a low molecular weight glycolipid isolated from the yeast-like fungus Pseudozyma flocculosa, and biopeptides from yeasts, in particular biopeptides from the yeast Metschnikowia reukaufii, isolated from the porcelain flower Hoya camosa.

[0039] The quantity of additional active ingredients in the topical composition usable according to the invention may vary depending on the nature of the ingredient used and the desired effect. Those skilled in the art shall ensure that the presence of this additional active ingredient does not alter the intrinsic properties of the tetrapeptide of the formula defined above.

[0040] The topical composition usable according to the invention can be presented in the various galenic forms classically used for topical application, i.e. in the form of a gel, in particular a superfatted cleansing gel or foaming gel, lotion, emulsion (in particular cream or milk), serum, balm, in particular anhydrous, oil, in particular soothing bath oil, spray, aerosol, bar, mask or ointment, containing usual physiologically acceptable excipients.

[0041] These forms of presentation for topical use can be prepared by known techniques, and for example, in the case of a cream, by dispersing a fatty phase in an aqueous phase to obtain an oil-in-water emulsion, or conversely by dispersing an aqueous phase in a fatty phase to prepare a water-in-oil emulsion.

[0042] Physiologically acceptable excipients may include, but are not limited to: - emulsifiers and co-emulsifiers such as sorbitan olivate, glyceryl stearate-citrate, coco-glucoside and glyceryl stearate, - consistency agents such as beeswax, glycol palmitate, and microcrystalline wax, - emollients such as silicone oils (e.g., dimethicone, caprylyl methicone, etc.), mineral oils (e.g., hydrogenated polyisobutene, paraffin oil, isohexadecane, etc.), isononyl isononanoate, lanolin, cetyl palmitate, palmitic acid, Ci0-Ci8 triglycerides, petrolatum, and vegetable oils such as sweet almond (Prunus dulcis) oil, wheat germ (Triticum vulgare) oil, and hemp (Cannabis) oil. sativa), and vegetable butters such as cocoa butter (Theobroma cacao) and mango butter (Mangifera indica L.), - Penetrating agents such as ethoxydiglycol, methylal, octyl dodecanol and propylene glycol, - humectants such as butylene glycol and polyethylene glycol and glycerin ethers such as glycereth-26, urea, - antioxidants such as vitamin C and its derivatives, vitamin E, vitamin F, vitamin PP (niacin and nicotinamide), - chelating agents such as phytic acid and ethylenediaminetetraacetic acid, - surfactants, particularly anionic ones, such as the disodium salt of glutamic acid amide and coco acid, sodium lauryl ether sulfate, and sodium olefin sulfonate in Ci4_i6, - preservatives such as phenylethyl alcohol, hydroxyacetophenone, chlorphenesin and sodium benzoate, - gelling agents such as polymers, in particular polyacrylates (carbomers), polyacrylamides, sclerotium gum, bentone, cellulose derivatives such as hydroxyethylcellulose, and chitosan derivatives, - pH modifying agents such as citric acid and sodium hydroxide, - solubilizers such as PEG 40 hydrogenated castor oil and ethoxylated sorbitan monolaurate (20 moles), - colorings such as beta-carotene, caramel, spirulina extract, orcanette root extract (Alkanna tinctoria), mica, iron oxide, etc.

[0043] The topical composition usable according to the invention is particularly well tolerated by the skin, exhibits no cytotoxicity, and its application to the skin, even for long periods, does not cause any systemic effects. It can be applied to the skin and / or mucous membranes, particularly to areas that require it, for example, once or twice a day.

[0044] The duration of application will vary depending on the improvement of the skin in the treated areas. As a guideline, the topical composition is preferably applied between flare-ups, and the duration of treatment can vary from approximately 8 to 30 days.

[0045] Other features and advantages of the present invention will become apparent from the following examples and the accompanying figures in which:

[0046] [Fig.1] is a schematic representation of a reconstructed skin model used in Example 1. EXAMPLES

[0047] The following products were used in the examples: - Tetrapeptide Pal-KTFC 6 ppm sold under the reference WGPALKTFKMBAL by the company Sederma; - Sodium dodecyl sulfate (reference L4509, Sigma) - Thapsigargine (reference T7459, Invitrogen TM) - ELISA kit for the assay of TSLP sold under the reference ELH-TSLP-1 by Raybiotech;

[0048] - Basal culture medium for culturing primary keratinocytes, free of components of animal origin, chemically defined and supplemented with a non-animal serum.

[0049] These ingredients were used as received from the manufacturers, without further purification.

[0050] EXAMPLE 1: Demonstration of the properties of the tetrapeptide Pal-KTFK for the prevention and / or treatment of atopic dermatitis

[0051] The study was conducted on a reconstruction of human epidermis. In vitro reconstructed human skin equivalents closely mimic normal human skin. Since these models largely reproduce the barrier function properties of normal human skin, they can be used to screen for potential skin irritants and to study effects related to product efficacy.

[0052] The tests presented in this example were carried out on an AlternaSensiSkin ® model. This model comprises normal human epidermal keratinocytes NHEK, one of the main cell types of the skin, cultured on polycarbonate inserts.

[0053] Figure 1 is a schematic profile (Fig. 1a) and vertical sectional (Fig. 1b) representation of the AlternaSensiSkin model used. This model (1) comprises at least one culture well (2) containing a basal culture medium (3) in which a culture insert (4) is placed. The insert has a membrane (5) at its base on which the human epidermal reconstruction (6) rests. The products to be tested (7) can be applied locally directly to the surface of the human epidermal reconstruction (6), and their irritant potential can be evaluated using a number of parameters, including cell viability, measurements of biological markers, or by studying the morphological components of the tissue.

[0054] The AlternaSensiSkin® model is derived from the AltemaSkin® model, which is an in vitro reconstruction of the human epidermis (RhE) from the company Cell Altemativ (adapted from the method of Poumay et al. (Arch. Dermatol. Res., 2004, 296: 203-211)). The AlternaSensiSkin® model is, in fact, an "immature" reconstructed AlternaSkin® epidermis designed to respect the morphology and properties of infant skin and / or sensitive skin, particularly the stratum corneum. It has been used in this An example is used as a model of atopic skin after incubation of reconstructed skin with thapsigargin. Indeed, TSLP secretion is highly dependent on calcium ions. Treatment with thapsigargin (TG) promotes the depletion of intracellular calcium reserves and induces TSLP secretion, which is abundantly produced by keratinocytes in cases of atopic dermatitis. 1.2 Samples tested

[0055] The different samples tested are presented in Table 1 below:

[0056] [Tables 1] Sample Description: Thapsigargin 1 pM (TG) Positive control for inflammation (atopic dermatitis) PAL-KTFK Tetrapeptide 3% Pal-KTFK in solution in a mixture of 75 pL 37% HCl and 5 mL SDS 2% dimethyl sulfoxy Positive control for the MTT test 1.3 MTT cytotoxicity test

[0057] The MTT test allows for the measurement of cellular metabolic activity, an indicator of cell viability, proliferation, and toxicity. This colorimetric assay is based on the reduction of a yellow tetrazolium salt [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide or MTT] into purple formazan crystals by metabolically active cells.

[0058] The tetrapeptide Pal-KTFK 6 ppm was tested at a concentration of 3% on reconstructed epidermis previously sensitized with thapsigargin at 1 pM.

[0059] To this end, treatment with 1 pM thapsigargin was carried out in the basal culture medium of RhE (n=4 per sample) for 42 h at 37°C. After this treatment time, the culture medium containing 1 pM thapsigargin was replaced with fresh culture medium without thapsigargin. The test product (20 pL) was applied directly to the surface of the epidermis and then incubated at 37°C for 20 hours. A positive control (SDS 2%) was performed under the same conditions, as well as a negative control without a sample.

[0060] At the end of the incubation period, the tissues were placed in a 0.5 mg / mL MTT solution in the culture medium for 3 h, then extracted in 2 x 500 pL of isopropanol. Cell viability was assessed by spectrophotometry at 570 nm using a Tristar2® microplate reader sold by Berthold Technologies GmbH, and absorbance values ​​were recorded using ICE® software version 1.0.9.8. Statistical analysis was performed using Graph Pad Prism v5.03 software.

[0061] Each measurement was taken in triplicate. 1.4. TSLP Assay

[0062] Treatment with 1 pM thapsigargin was carried out in the basal culture medium of RhE (n = 4 per sample) for 48 hours at 37°C to induce TSLP secretion. After this treatment time, the culture medium with thapsigargin was replaced with fresh culture medium. The test product (20 pL) was then applied directly to the epidermal surface and incubated at 37°C for 20 hours. A positive thapsigargin control was performed under the same conditions for 42 hours with thapsigargin but without application of the test product for the following 20 hours.

[0063] A negative (untreated) control was carried out under the same conditions but without application of thapsigargin or product to be tested.

[0064] TSLP was quantified using the ELH-TSLP-1 ELISA kit to determine the amount produced by reconstructed epidermis after various thapsigargin treatments. For this purpose, cell supernatants were collected after the required treatment time (42 hours), and a quantitative assay was performed using the kit according to the manufacturer's recommendations. The procedure involves loading the samples and a range of standards into the wells of a duplicate 96-well microplate. The TSLP marker present in a sample is bound by the antibody immobilized at the bottom of the well. The wells are then washed, and a biotinylated antibody is added. After washing, HRP-streptavidin is loaded into the wells. The wells are then washed again, and a substrate solution is added to the wells, resulting in a blue color whose intensity is proportional to the amount of bound TSLP marker.A development solution is used to change the color from blue to yellow, and the intensity of the staining is measured at a wavelength of 450 nm using a Tristar2® microplate reader sold by Berthold Technologies GmbH. Absorbance values ​​are recorded and analyzed using ICE vl.0.9.8 software. The result is interpreted as a sample / untreated ratio. A value greater than or equal to 2 indicates TSLP overexpression. 1.5 Results

[0065] The results of the cell viability test are presented in Table 2 below: (TABLE 2] Ecnant-iîon ' VULHity Vîata te * * i Standard deviation (%) i Untreated control 13. i 2 es y Positive test (BBS 2%} ] . 7 18________________________________________________[ ThspUqsrqme control 42 H jh 7 2 - LO i PAL-KTFK 3 % 42 H i 64.9 S 54 Ü * : Mayenne of the 3 trials

[0066] The results presented in Table 2 show that prior treatment with thapsigargin leads to significant sensitization of the epidermis, indicative of a state of atopic dermatitis. Subsequent application of the PAL-KTFK peptide to the previously thapsigargin-sensitized epidermis does not result in a significant decrease in cell viability, which decreases from 67.2% to 64.9%, representing a reduction of only about 3.4%. Therefore, the PAL-KTFK tetrapeptide used according to the present invention does not exhibit a significant cytotoxic effect on the cells of the reconstructed epidermal model used in the test described in this example.

[0067] The results of the TSLP assay are presented in Table 3 below: [Tables3] Sample TSLP Concentration (pg / ml) Untreated Control 0.29 Thapsigargin Control 2.44 Thapsigargin + tetrapeptide PAL-KTFK 6 ppm at 3% 1.30

[0068] The results presented in Table 3 show that contact of the reconstructed epidermis with thapsigargin leads to overexpression of the cytokine TSLP, involved in the phenomenon of atopic dermatitis (8.4 for the untreated control). Treatment of the reconstructed epidermis for 20 hours with the tetrapeptide PAL-KTFK according to the invention results in a significant decrease in TSLP overexpression compared to the thapsigargin control (-1.14 pg / mL), i.e., by a factor of 1.87.

[0069] All of these results demonstrate that the tetrapeptide Pal-KTFK has advantageous properties for use in the prevention and / or treatment of atopic dermatitis. EXAMPLE 2#: Preparing a body lotion

[0070] In this example, a body lotion having the following composition was prepared: Phase A: - Combination of microcrystalline cellulose (MCC) and xanthan gum (XG) and cellulose gum (CMC), sold under the trade name Vivapur CS Tex Easy ® by the company JRS Rettenmaier 3% - Butylene glycol 5% - Demineralized water 60.2% Phase B: - Paraffin oil 8% - Mixture of microcrystalline wax, paraffin oil and paraffin sold under the trade name Vaseline Codex P23 by the company Aiglon 5% - Isopropyl palmitate (STEARINERIE DUBOIS) 5% - Mixture of glyceryl stearate and PEG-100 stearate (Seppic) 5% Phase C: - Dimethicone (KCC Beauty) 1% Phase D: - Tetrapeptide PAL-KTFK formulated as an emulsion, sold under the trade name Crystalide® by Sederma 3% - Mixture of hydrogenated phospholipids, ethylene glycol, glycerin, calcium chloride dihydrate and demineralized water sold under the trade name Epicalsome® by Mibelle Biochemistry 2% - Pentylene glycol 1% - Sodium benzoate 0.5% - Potassium sorbate 0.5%

[0071] The components of phase A were weighed and then stirred at 75°C. The components of phase B were weighed and then stirred at 75°C. The emulsion was prepared by pouring phase B into phase A while stirring vigorously, then cooling it to a temperature of approximately 50°C before adding phase C. The components of phase D were then successively added to the mixture thus obtained after it had been cooled to a temperature of approximately 35°C.

[0072] The pH of the body milk thus obtained was adjusted to a value between 4.8 and 5.0, using a 20% citric acid solution. EXAMPLE 3#: Preparing a body balm

[0073] In this example, a body balm having the following composition was prepared:

[0074] Phase A: - Butylene glycol 5% - Mixture of sodium acrylate / sodium acryloyldimethyltaurate copolymer, isohexadecane and polysorbate 80 sold under the Trade name Simulgel ® EG by the company Seppic 3% - Demineralized water 56.87% Phase B: - Mixture of microcrystalline wax, paraffin oil and paraffin sold under the trade name Vaseline Codex Diamant N by the company Aiglon 10% - Capric / caprylic triglycerides (BASF) 10% - Glyceryl stearate 1.8% - Cetearyl alcohol 1.5% - Myristyle myristate 2% - Sodium stearoyl glutamate 1% Phase C: - Dimethicone 1% Phase D: - Tetrapeptide PAL-KTFK in solution in a mixture of glycerin and water, sold under the reference WG PAL KTFK MB AL by the company Sederma 3% - Mixture of hydrogenated phospholipids, ethylene glycol, glycerin, calcium chloride dihydrate and demineralized water sold under the trade name Epicalsome ® by the company Mibelle Biochemistry 2% - Sodium benzoate 0.50% - Potassium sorbate 0.50%

[0075] The components of phase A were weighed and then stirred at 75°C. The components of phase B were weighed and then stirred at 75°C. The emulsion was prepared by pouring phase B into phase A while stirring vigorously, then cooling it to a temperature of approximately 50°C before adding phase C. The components of phase D were then successively added to the mixture thus obtained after it had been cooled to a temperature of approximately 35°C.

[0076] The pH of the body balm thus obtained was adjusted to a value between 4.8 and 5.0, using a 20% citric acid solution. EXAMPLE 4#: Preparing an eyelid treatment

[0077] In this example, an eyelid treatment having the following composition was prepared: Phase A#:

[0078] - Glycerin 2% - Carbomer 0.5% - Demineralized water 78.52% Phase B: - Dicaprylyl carbonate 5% - Shea butter 2% - Cetyl alcohol 1.5% - Potassium cetyl phosphate 2% Phase C: - Dimethicone 1% Phase D: - Tetrapeptide PAL-KTFK formulated as an emulsion, sold under the trade name Crystalide® by the company Sederma 3% - Pentylene glycol 1% - Chlorphenesin 0.2% - Phenoxyethanol 0.6% The components of phase A were weighed and then stirred at 75°C. The components of phase B were weighed and then stirred at 75°C. The emulsion was prepared by pouring phase B into phase A while stirring vigorously, then cooling it to a temperature of approximately 50°C before adding phase C. The components of phase D were then successively added to the resulting mixture after it had been cooled to a temperature of approximately 35°C.

[0079] The pH of the eyelid treatment thus obtained was adjusted to a value between 6.5 and 7.5, using a 10% sodium hydroxide solution. EXAMPLE 5#: Preparing a foaming gel

[0080] In this example, a foaming gel having the following composition was prepared: Phase A: - Demineralized water 81.76% Phase B: - Xanthan gum 1.1% - Glycerin 2.0% Phase C: - Tetrapeptide PAL-KTFK formulated as an emulsion, sold under the trade name Crystalide® by the company Sederma 1% - Decyl glucoside 8% Phase D: - Phenoxyethanol 5% - Chlorphenesin 0.28% Phase E: - 50% citric acid solution 0.26%

[0081] The components of phase B were pre-dispersed, and then phase B was added to phase A while stirring until a homogeneous mixture was obtained. The components of phase C were mixed until a homogeneous mixture was obtained, and then phase C was added to the mixture of phases A and B. Finally, the components of phase D were mixed until a homogeneous mixture was obtained, and then phase D was added to the previously obtained mixture of phases A, B, and C.

[0082] The pH was adjusted if necessary between 5 and 6 using phase E. EXAMPLE 6#: Preparing a washing oil

[0083] In this example, a washing oil having the following composition was prepared: Phase A: - Sodium lauryl sulfate in aqueous solution sold under the trade name TEXAPON NSO UP by BASF 35% - Cocamidopropyl betaine and sodium chloride in aqueous solution sold under the trade name DEHYTON K by BASF 10% - Sodium salt of cottonseed oil amphoacetate sold under the trade name SHAROSURF CN by SHARON 5% - Tetrapeptide PAL-KTFK in solution in a mixture of glycerin and water, sold under the reference WG PAL KTFK MB AL by Sederma 1% - Cottonseed oil 0.2% Phase B: - Glycerin 15% - Phenoxyethanol 0.6% - Chlorphenesin 0.25% - Demineralized water 32.83% PHASE C: - 50% aqueous citric acid solution 0.13%

[0084] The components of phase A were mixed with stirring. The compounds of phase B were mixed with stirring. Phase B was then slowly added to phase A with gentle stirring until a homogeneous mixture was obtained. The pH was adjusted, if necessary, between 5 and 6 using phase E.

Claims

Demands

1. Tetrapeptide of formula N-Palmitoyl-Lys-Thr-Phe-Lys, for its use as an active ingredient for the prevention and / or treatment of human atopic dermatitis.

2. Topical composition comprising at least one physiologically acceptable excipient and, as an active ingredient, at least one tetrapeptide of formula N-palmitoyl-Lys-Thr-Phe-Lys, for use in the prevention and / or treatment of human atopic dermatitis.

3. Topical composition for use according to claim 2, characterized in that the content of said tetrapeptide within said composition is 2 to 8 ppm, relative to the total weight of said topical composition.

4. Topical composition for use according to claim 2 or 3, characterized in that said tetrapeptide is used in the form of an emulsion.

5. Topical composition for use according to claim 4, characterized in that said emulsion is an oil-in-water emulsion comprising one or more fatty acid triglycerides.

6. Topical composition for use according to any one of claims 2 to 5, characterized in that said composition further comprises calcium ions.

7. Topical composition for use according to any one of claims 2 to 6, characterized in that said calcium ions are encapsulated in a supramolecular structure comprising one or more concentric lipid bilayers trapping between them aqueous compartments comprising said calcium ions.

8. Topical composition for use according to claim 6 or 7, characterized in that the calcium ion content is 13 to 32 ppm relative to the total weight of said composition.

9. Topical composition for use according to any one of claims 2 to 8, characterized in that said composition further comprises one or more additional active ingredients selected from anti-pruritic actives, cutaneous immune system rebalancing actives and actives acting on cutaneous microbiome dysbiosis.

10. Topical composition for use according to claim 9, characterized in that the anti-pruritic active ingredients are selected from thermal waters, saccharidic gums obtained by fermentation of sorbitol, and plant extracts of Boswellia serrata, Acmella oleracea, Aloe vera, oat (Avena sativa), burdock (Arctium iappa), calendula (Calendula), German chamomile (Matricaria recutita), fumitory (Fumaria officinalis), witch hazel (Hamamelis virginiana), lavender (Lavandula officinalis or angustifolia or vera), mallow (Malva sylvestris), lemon balm (Melissa officinalis), stinging nettle (Urtica dioica or Urtica urens), wild pansy (Viola tricolor), horsetail (Equisetum arvenses), sarsaparilla (Smilax aspera), tea tree (Melaleuca altemifolia), white birch (Betula alba), red sage (Salvia miltiorrhiza), or heart pea (Cardiospermum halicacabum);hydrolysates of corn or oat starch, and oily active ingredients selected from peroxidized corn oil (Zea mays) and bisabolol.

11. Topical composition for use according to claim 9, characterized in that the cutaneous immune system rebalancing actives are selected from unsubstituted [3-glucans and [3-glucans substituted with carboxymethyl groups.

12. Topical composition for use according to claim 9, characterized in that the active ingredients acting on skin microbiome dysbiosis are selected from alpha-glucan oligosaccharides, Yacon tuber extract (Polymnia sonchifolia), Lactobacillus, rhamnose, glucose, glucuronic acid, glycolipids, biopeptides from the yeast Metschnikowia reukaufii, isolated from the Hoya camosa porcelain flower.

13. Topical composition for use according to any one of claims 2 to 12, characterized in that it is in the form of a gel, lotion, emulsion, serum, balm, oil, spray, aerosol, bar, mask or ointment.

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