CZ309857B6 - Hexapeptide, compositions comprising this hexapeptide, and topical use thereof - Google Patents

Abstract

The solution relates to a hexapeptide as a skin cell enhancer with the general formula of I X-Phe-Trp-Ala-His-Lys-Lys-Z (I) wherein: X is an -NHX1 phenylalanine group at the N-terminal end of the hexapeptide, wherein X1 is selected from H, acetyl, octanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, elaidoyl, oleoyl, biotinoyl, or lipoyl; Z is a -COZ1 group of lysine at the C-terminal end of the hexapeptide, wherein Z1 is selected from the group consisting of OH, OCH3, OCH2CH3, or NH2; and topical compositions comprising this hexapeptide and their topical use.

Description

Hexapeptide, compositions comprising this hexapeptide, and topical use thereof

Field of technology

The present invention relates to an engineered peptide of the sequence X-Phe-Trp-Ala-His-Lys-Lys-Z, its derivatives or complexes with metal ions, a topical composition comprising them and their topical use. It relates in particular to the use of peptides for treating the skin of humans or animals for cosmetic or dermo-pharmaceutical purposes.

Current state of the art

Acne vulgaris

Acne vulgaris (hereafter acne) is a very common chronic skin disease of the pilosebaceous unit with a profound negative psychosocial impact on patients' quality of life. Although it affects individuals of all ages, it usually appears during adolescence and often persists into adulthood (Yentzer et al. 2010).

The pathogenesis of acne vulgaris is multifactorial with four major events: hyperseborrhea, epithelial hyperproliferation and hyperkeratinization, Cutibacterium acnes colonization, and inflammation.

Hyperseborrhea

Increased sebum production is considered one of the key events in the pathogenesis of acne. Different signaling pathway compounds are involved in the regulation of sebum production by the skin, of which six hormones play a key role. Among the most important stimulators of sebum production are androgens, the upregulation of which is often associated with the development of acne (da Cunha, Fonseca, and Machado 2013). In skin cells, testosterone, the most common androgen, is rapidly reduced to 5α-dihydrotestosterone (DHT) by 5α-reductase, type I (Fritsch, Orfanos, and Zouboulis 2001). Both testosterone and DHT bind to androgen receptors (ARs), but DHT is a 5- to 10-fold more potent AR agonist and is considered the major androgen hormone in hair follicles (Anderson and Liao 1968).

Epithelial hyperproliferation and hyperkeratinization

Another typical event in the pathogenesis of acne is the hyperproliferation of follicular keratinocytes and their abnormal differentiation (Thiboutot 2000). The latter is associated with increased cohesiveness of corneocytes, which leads to their non-functional desquamation (Thiboutot 2000). The accumulated mass of keratinized corneocytes together with the sebum partially obstructs the follicles, which leads to their stretching and the creation of suitable conditions for the growth of C. acnes (Thiboutot 2000).

There are several hypotheses explaining the cause of keratinocyte hyperproliferation and altered differentiation in acne. Androgens, in addition to stimulating sebum production, have been shown to induce the aforementioned processes as well as the production of the pro-inflammatory interleukin IL-1α (Kumtornrut et al. 2019; Guy and Kealey 1998). Another possible trigger for hyperkeratinization is a low concentration of the essential fatty acid, linolenic acid, which is reduced in acne-affected skin due to its dilution by high levels of sebum (Downing et al. 1986).

Cutibacterium acnes

C. acnes is a bacterium living on and in human skin as part of the normal human skin microbiome. It resides mainly deep in sebaceous follicles in contact with keratinocytes.

- 1 CZ 309857 B6

C. acnes is confirmed to play a significant role in the pathogenesis of acne (McLaughlin et al. 2019). It seems that the total amount of C. acnes is not as important for the development of acne as the presence of certain strains of C. acnes (Dréno et al. 2018). These strains associated with acne have been shown to be more virulent and produce a large number of different pro-inflammatory factors (Dréno et al. 2018).

Inflammation

Inflammation is considered a key part in the pathogenesis of acne. In the past, inflammation was considered a secondary event induced by C. acnes colonization. Inflammatory processes are now believed to be involved in all stages of acne development, and acne is now considered a truly inflammatory disease (Tanghetti 2013).

Acne treatment

Because acne is a multifactorial disease, a combination therapy approach is recommended. A combination of a topical retinoid (tretinoin, isotretinoin, adapalene, tazarotene, retinol, retinaldehyde, etc.) and an antimicrobial agent (eg, benzoyl peroxide) is typically chosen as first-line treatment (Moradi Tuchayi et al. 2015). Hormonal, antiandrogen therapy (oral contraceptives, spironolactone) is often given to women to reduce sebum production. Topical and oral antibiotics (erythromycin, clindamycin, doxycycline, minocycline, and sarecycline) are used to treat mild to severe acne, usually in combination with benzoyl peroxide or retinoids. Other anti-acne surfactants used include salicylic acid, azelaic acid, zinc, sulfur, niacinamide, glycolic acid, and antimicrobial peptides (Liu et al. 2020).

Zinc in the treatment of acne

Zinc levels in acne patients are often significantly lower, and zinc supplementation (orally or topically) has been shown to improve acne (Yee et al. 2020). Although its exact mechanism of action is not fully understood, current knowledge suggests multiple mechanisms including anti-inflammatory, antioxidant effects, inhibition of C. acnes proliferation and inhibition of 5n-reductase, leading to reduced sebum production in vivo (Abendrot and Kalinowska-Lis 2018; Cervantes and col. 2018).

Peptides and acne treatment

Another promising group of compounds in the treatment of acne are peptides showing antimicrobial (AMP) (Ma et al. 2021; Melo, Dugourd, and Castanho 2006; Zhang et al. 2013), anti-inflammatory (Zhang et al. 2013) and other properties such as antioxidant effects (Mills et al. 2016). Some naturally derived or purely synthetic substances designed by AMPs have been shown to have promising anti-acne effects:

• Omiganan pentahydrochloride derived from the bovine AMP indolicidin (Melo, Dugourd, and Castanho 2006; Rubinchik et al. 2009) • GDP-20 (peptides derived from granulysin) (Lim et al. 2015; Ma et al. 2021;

McInturff, Modlin, and Kim 2005) • Peptide LZ1 (VKRWKKWWRKWKKWV-NH2) (Zhang et al. 2013) • AMPs from frog skin ([D4k]ascaphin-8, [G4K]XT-7, [T5k]temporin-Dra, brevinin-2GU and B2RP-Era (Popovic et al. 2012) • Peptides (Br-p) isolated from burdock root (Arctium lappa L.) (Miazga-Karska, Michalak and Ginalska 2020)

- 2 CZ 309857 B6 • Temporin-1Dra and moronecidin (Wu, Zhang and Zhou 2020) • Bombinin-like peptide 7 (BLP-7) from Bombina orientalis (Wu et al. 2020) • Cathelicidin-BF was purified from snake venoms from Bungarus fasciatus (Wang et al. 2011) • Head-to-tail cyclized bacteriocin AS-48 (Cebrián et al. 2018) • CEN1HC-Br isolated from green sea cucumber (Han et al. 2018) • AMP C12-50 (Nair et al. al. 2017) • α-helical cationic peptide, P5 (Sunhyo Ryu et al. 2015) • Helicobacter pylori - derived synthetic antimicrobial peptide HPA3NT3 (S. Ryu et al. 2014) • Synthetic epinecidin-1(22-42) peptide was derived from positions 22-42 of Epinephelus coioides epinecidin-1 (Pan et al. 2009) • Enterococcus faecalis-derived AMP SL-5 (Kang et al. 2009) • Bacteriocin-like inhibitory substance (BLIS-like substance) produced by Streptococcus salivarius (Bowe et al. 2006) • Synthetic, designed by AMP (Woodburn, Jaynes, and Clemens 2020)

Examples of patents describing the use of peptides in the treatment of acne:

• WO 2021175186 A1 Use of a small molecule short peptide in the formulation of an acne treatment product (LQLWQAEER, derived from bFGF, 100 μg/ml, inhibits sebocyte growth, reduces sebum production) • CN 111253469 A Self-assembly short chain peptide and application of short chain peptide in the treatment of acne (FFLQLQAEER) • CN 107698662 A Peptide combination for the treatment of acne (WKIKIDSEAE and

PNMIYSKDY) • CN 112979762 A Cyclic peptide PIZ and its application (EYPYKHSGYYHRAV) • KR 20200101767 A Peptide with antimicrobial activity and composition for antimicrobial comprising the same (KTTKS) • WO 03015809 A2 Antimicrobial cationic peptides and their formulation • WO 2005025598 A1 Antibacterial drug for Propionibacterium acnes • EP 1853620 A2 Antimicrobial hexapeptides

However, there is no patent or publication describing any peptide of the amino acid sequence of the present invention, which is key to the physicochemical properties of the peptide as well as its biological activity.

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Many of the treatment options mentioned above have negative side effects. Retinoids in particular, but also zinc or salicylic acid, commonly cause skin dryness, peeling, itching, redness, and sun sensitivity (Thielitz and Gollnick 2008; Cervantes et al. 2018; Arif 2015). Of particular concern is the known teratogenic effect of retinoids observed after oral use (Thielitz and Gollnick 2008). Although topical retinoids have not been shown to increase the risk of birth defects, they are not recommended for pregnant women as a precaution (Panchaud et al. 2012). The use of antibiotics as another common approach in acne therapy is associated with a high risk of selection of resistant bacterial strains (Walsh, Efthimiou and Dréno 2016). Therefore, they cannot be used as monotherapy and are only recommended for short-term use (Walsh, Efthimiou and Dréno 2016).

For all these reasons, there is still a need for new, more effective, comprehensive and safer acne treatments.

The essence of the invention

Surprisingly, novel activities of the hexapeptide Phe-Trp-Ala-His-Lys-Lys on skin cells and improvement of skin condition were found.

The subject of the invention relates to a hexapeptide as a skin cell improver with the general formula I

X-Phe-Trp-Ala-His-Lys-Lys-Z (I) where:

X is a -NHX ¹ phenylalanine group at the N-terminal end of the hexapeptide, where

X ¹ is selected from the group consisting of H, acetyl, octanoyl, decanoyl, lauryl, myristoyl, palmitoyl, stearoyl, elaidoyl, oleoyl, biotinoyl or lipoyl. Preferably X ¹ is H.

Z is a COZ ¹ lysine group at the C-terminal end of the hexapeptide, where Z ¹ is selected from the group consisting of OH, OCH 3 , OCH 2 CH 3 or NH 2 . Preferably, Z ¹ is OH.

The hexapeptide of the present invention improves skin cells so that it can be applied topically to the skin and can also be called a topical hexapeptide.

The hexapeptide of general formula I according to the present invention is preferably in the form of a complex with a metal ion selected from the group comprising a zinc ion, a copper ion, a manganese ion or a magnesium ion. The metal ion is bound by a coordinative covalent bond in the complex. Preferably, the metal ion is zinc ion (Zn ²⁺ ) and the complex is shown schematically below as the Zn-FWAHKK complex or simply as Zn-FWAHKK.

The hexapeptide of general formula I according to the present invention can be optically pure or composed of L- or D-isomers or their mixture. The L-isomers, which are those found in nature, are preferred.

The hexapeptide according to the present invention can be in the form of salts, especially salts of hydrochloric acid, formic acid or acetic acid, or any salt commonly used in cosmetics.

Another embodiment according to the present invention is a topical composition comprising at least one hexapeptide of general formula I as mentioned above.

- 4 CZ 309857 B6

According to a preferred embodiment of the topical composition according to the present invention, the concentration of the hexapeptide of general formula I, as mentioned above, is in the range of 0.0001 to 0.135% (w/w), preferably in the range of 0.001 to 0.05% (w/w wt.), more preferably from 0.001 to 0.0135% (wt/wt). The amount of hexapeptide depends on the purpose of the composition and the desired final effect.

The present invention also includes a topical composition that contains at least one hexapeptide according to the present invention in a form for topical skin application, which can be selected from the group consisting of cream, serum, anhydrous gel, paste, vesicle dispersion, powder, nanofibers, macro-, micro- or nanoparticles or macro-, micro- or nano-spheres that can be adsorbed on organic polymer powders, talcs, bentonites and other inorganic or organic support materials.

This topical composition according to the present invention is in the form of a cream selected from the group comprising a water-in-oil or oil-in-water emulsion, micro- or nano-emulsion.

This topical composition according to the present invention is in the form of a serum selected from the group comprising a sterile or non-sterile aqueous or aqueous-alcohol solution or gel.

According to another preferred embodiment, the topical composition according to the present invention is in a form selected from the group comprising a cream or a serum.

In the case that the topical composition of the present invention is in the form of a cream, the concentration of the hexapeptide of the general formula I as mentioned above is in the range of 0.0001 to 0.135% (w/w), preferably 0.001% to 0.0135 % (w/w).

Advantageously, the composition in the form of a cream further comprises at least one cosmetic or dermopharmaceutical excipient selected from the group comprising oil, wax, butter, emulsifier, auxiliary active ingredient or preservative.

Preferably, the amount of oil, wax, butter or their mixture in the composition according to the present invention is in the range of 20 to 55% (wt/wt), preferably 20 to 40% (wt/wt), more preferably 20 to 30% (wt/wt). Preferably, the amount of emulsifier in the composition according to the present invention is in the range from 1 to 20% (w/w), preferably 2 to 15% (w/w), more preferably 2 to 10% (w/w). ). Preferably, the amount of the additional active ingredient in the composition according to the present invention is in the range from 0.001 to 20% (wt/wt), preferably 0.001 to 10% (wt/wt), more preferably 0.001 to 5% (wt/wt wt.). Preferably, the amount of preservative in the composition according to the present invention is in the range from 0.1 to 2.5% (wt/wt), preferably 0.1 to 1.5% (wt/wt), more preferably 0, 8 to 1.2% (w/w).

Preferably, the oil is selected from the group including argan, coconut, avocado, almond, sesame, olive, sunflower, hemp, jojoba, macadamia, wheat germ, marula, safflower, rice, poppy, rosehip, apricot, castor oil, caprylic/carp oil triglyceride; the butter is selected from the group consisting of cocoa butter, shea butter, ilip butter, kokum butter, murumuru butter, mango butter, kupuak butter, avocado butter; and the wax is selected from the group consisting of lanolin, beeswax, carnauba, candelilla wax and petroleum jelly.

Advantageously, the emulsifier is selected from the group comprising glyceryl stearate, glyceryl caprylate, behenyl alcohol, glyceryl behenate, cetearyl glucoside, methyl glucose, sestearate, glyceryl stearate citrate, polyglyceryl-3 stearate, cetearyl olivate, lecithin, stearyl alcohol, sorbitan oleate, polysorbate, stearic acid, cetyl alcohol and cetearyl alcohol, sodium acrylate, sodium acryloyldimethyl taurate copolymer, isohexadecane, polysorbate 80 or a mixture thereof.

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A topical composition according to the present invention in the form of a cream containing at least one hexapeptide according to the present invention may also be combined with other auxiliary active ingredients that may have a synergistic or additive effect to increase or extend the desired activity described in the present invention. This additional active ingredient is selected from the group containing the following: anti-acne, anti-aging, anti-wrinkle, lightening, whitening, anti-blemish, pigmenting, hydrating, moisturizing, moisturizing, slimming, exfoliating, anti-redness, anti-inflammatory, antioxidant, radical scavenger, anti-glycation, increasing volume, restructuring, rejuvenating, regenerating, anti-carbonylation, dermo-relaxing, improving the stratum corneum, dermo-epidermal connection, firmness, elasticity, collagen boosters, eye contours (dark circles and bags under the eyes), promoting blood circulation, etc.

The other active ingredients mentioned above can be synthetic or obtained from plants, bacteria, fungi, cell cultures or their products.

Advantageously, another active ingredient is selected from the group comprising vitamins A, D, E, K, C, Bgroup, coenzyme Q10, allantoin, bisabolol, bakuchiol, resveratrol, lactic acid, amino acids, benzoyl peroxide, sulfur, azelaic acid, peptides, preferably acetyl hexapeptide-8, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, copper tripeptide-1, hexapeptide-1, palmitoyl pentapeptide-4, saccharomyces peptides and proteins, preferably rice, soy, quinoa or wheat protein; polysaccharides, preferably hyaluronic acid or its pharmaceutically and cosmetically acceptable salt or derivative, preferably sodium oleoyl hyaluronate, sodium retinoyl hyaluronate, crosspolymer-3 sodium hyaluronate; carboxymethyl glucan, schizophyllan, glucomannan; panthenol; urea; glycerine; plant extracts, preferably aloe vera extract, chamomile extract, acai extract, green tea extract, algae extract, oat extract, hemp extract, cranberry extract; extracts, ferments, lysates or filtrates from bacteria, preferably from Lactobacillus spp., Thalassospira spp., Bifidobacterium spp., Halobacterium spp., or from fungi, preferably from Saccharomyces spp., Agaricus subrufencens, Choiromyces maendrformis, Cordyceps sinensis, Ganoderma lucidum, Grfola frondosa, Hypsizygus ulmarium, Inonotus obliquus, Lentinula edodes, Polyporus spp., Trametes versicolor, Tremella fucformis, Tuber spp., Schizophyllum commune.

Preferably, the preservative is selected from the group including aromatic acids and their derivatives, preferably benzoic acid, sodium benzoate, salicylic acid, dehydroacetic acid, potassium sorbate, parabens; alcohols preferably ethanol, isopropanol, benzyl alcohol, phenoxyethanol, phenethyl alcohol; imidazole derivatives preferably hydantoin, imidazolidinyl urea; cationic surfactants, preferably benzalkonium chloride.

When the topical composition of the present invention is in the form of a serum, the concentration of the hexapeptide of general formula I as mentioned above is in the range of 0.001 to 0.135% (wt/wt), preferably 0.001% to 0.0135% (wt/wt wt.).

Advantageously, the composition in the form of a serum further comprises at least one cosmetic or dermopharmaceutical auxiliary ingredient selected from the group comprising a thickener, an auxiliary active ingredient and a preservative as defined above.

Preferably, the amount of thickener in the composition according to the present invention is in the range of 0.1% to 20% (w/w), preferably 0.1% to 5% (w/w), more preferably 0.2 to 0 .5% (w/w). Preferably, the amount of auxiliary active ingredient in the composition according to the present invention is in the range of 0.001 to 20% (wt/wt), preferably 0.001 to 10% (wt/wt), more preferably 0.001 to 5% (wt/wt .). Preferably, the amount of preservative in the composition according to the present invention is in the range of 0.1 to 2.5% (wt/wt), preferably 0.1 to 1.5% (wt/wt), more preferably 0.8 up to 1.2% (w/w).

Preferably, the thickener is selected from the group consisting of xantham gum, sclerotia gum, guar gum, cellulose gum, carbomer, hydroxyethyl cellulose, root extract

- 6 CZ 309857 B6 from Amorphophallus konjac, carrageenan, pullulan, lecithin, lysolecithin, sodium acrylate, sodium acryloyldimethyltaurate copolymer, isohexadecane, polysorbate 80 or their mixtures.

A topical composition according to the present invention containing at least one hexapeptide according to the present invention may also be combined with other active ingredients that may have a synergistic or additive effect to increase the scope of the desired activity described in the invention. The auxiliary active ingredient is selected from the group including the following substances: anti-acne, anti-aging, anti-wrinkle, lightening, whitening, anti-blemish, pigmenting, hydrating, moisturizing, moisturizing, slimming, exfoliating, anti-redness, anti-inflammatory, antioxidants, radical scavengers, anti glycation, increasing volume, restructuring, rejuvenating, regenerating, anti-carbonylation, dermo-relaxing, improving the stratum corneum, dermo-epidermal connection, firmness, elasticity, collagen boosters, eye contours (dark circles and bags under the eyes), improving blood circulation, etc.

The auxiliary active ingredients mentioned above can be synthetic or obtained from plants, bacteria, fungi, cell cultures or their products.

Advantageously, the auxiliary active ingredient is selected from the group comprising vitamin C and the group of B vitamins; amino acids, peptides preferably acetyl hexapeptide-8, palmitoyl tripeptide-1, copper tripeptide-1, Saccharomyces peptides, hexapeptide-1, and proteins preferably rice protein, soy protein, quinoa or wheat protein; other polysaccharides, preferably hyaluronic acid or its pharmaceutically and cosmetically acceptable salt or derivative, preferably sodium oleoyl hyaluronate, sodium retinoyl hyaluronate, crospolymer-3 sodium hyaluronate; carboxymethyl glucan, schizophyllan, glucomannan, panthenol, urea, glycerin; plant extracts, preferably aloe vera extract, chamomile extract, acai extract, green tea extract, algae extract, oat extract, hemp extract, cranberry extract; extracts, ferments, lysates or filtrates from bacteria, preferably from Lactobacillus spp., Thalassospira spp., Bifidobacterium spp., Halobacterium spp., or mushrooms, preferably from Saccharomyces spp., Agaricus subrufescens, Choiromyces maendrformis, Cordyceps sinensis, Ganoderma lucidum, Grfola. frondosa, Hypsizygus ulmarium, Inonotus obliquus, Lentinula edodes, Polyporus spp., Trametes versicolor, Tremella fucformis, Tuber spp., Schizophyllum commune.

Preferably, the preservative is selected from the group comprising aromatic acids and their derivatives, preferably benzoic acid, sodium benzoate, salicylic acid, dehydroacetic acid; potassium sorbate, parabens; alcohols preferably ethanol, isopropanol, benzyl alcohol, phenoxyethanol, phenethyl alcohol; imidazole derivatives preferably hydantoin, imidazolidinyl urea; cationic surfactants, preferably benzalkonium chloride.

All % given below are (wt/wt%) unless otherwise stated and refer to the total weight of the composition when the % refers to the topical composition of the present invention.

In general, the amount of water in the composition of the present invention in the form of a cream or serum corresponds to the amount of water added to 100% (w/w) of the composition.

Cosmetically and pharmaceutically acceptable salts of hyaluronic acid should be those formed from acids that form non-toxic acidic anions selected from the group including sodium, potassium, calcium, magnesium, zinc.

New is the hexapeptide of general formula I according to the present invention and its use or the use of cosmetic or dermo-pharmaceutical topical compositions according to the present invention containing said hexapeptide for improving oily and acne-prone skin condition and appearance. More specifically, the hexapeptide of the present invention inhibits epidermal keratinization, reduces sebum production by inhibiting the key enzyme 5n-reductase, has anti-inflammatory activity by inhibiting pro-inflammatory interleukins, has an antimicrobial effect on Cutibacterium

- 7 CZ 309857 B6 acnes and reduces the number of acne lesions in vivo. Furthermore, said hexapeptide according to the present invention also has an anti-aging effect by stimulating collagen synthesis and reducing wrinkles in vivo.

Said hexapeptide of the topical composition according to the present invention is intended for oily and acne-prone skin as well as for normal skin.

According to another preferred embodiment, the hexapeptide according to the general formula I according to the present invention and the topical composition according to the present invention are intended for use in the treatment of the skin. Preferably for use in the medical treatment of skin diseases selected from the group including acne, atopic dermatitis, psoriasis, skin peeling disease, ichthyosis. Or preferably for use for cosmetic skin treatment.

Clarification of drawings

Giant. 1 Hexapeptide FWAHKK forms a complex with zinc. Spectrophotometric determination of the ability of zinc to form a complex with FWAHKK hexapeptide using dithizone as a free zinc indicator.

Giant. 2. Zinc penetrates cells more efficiently when it is in the Zn-FWAHKK complex. HaCaT keratinocytes were incubated with Zn-FWAHKK or ZnSO4*7H2O (Zn) at the appropriate concentration for 3 h. Intracellular zinc was visualized using fluorescence microscopy.

Giant. 3. Hexapeptide FWAHKK increases cell viability and prevents the cytotoxic effect of zinc in the Zn-FWAHKK complex. NIH-3T3 fibroblasts were incubated with FWAHKK, ZnFWAHKK or ZnSO4*7H2O (Zn) at the corresponding concentrations for 48 h. Cell viability was determined by the MTT method. *p<0.05; ** p<0.01; *** p<0.001 compared to control unless otherwise stated.

Giant. 4. Hexapeptide FWAHKK and ZnSO4*7H2O decreased 5n-reductase gene expression compared to retinol. The Zn-FWAHKK complex was even more effective than its individual components. HaCaT keratinocytes were treated with FWAHKK, Zn-FWAHKK, ZnSO4*7H2O (Zn) and retinol for 72 h. Relative gene expression of 5n-reductase (SRD5A1 gene) was determined by qRT-PCR. *p<0.05; ** p<0.01; *** p<0.001 compared to respective controls.

Giant. 5. Hexapeptides FWAHKK and Zn-FWAHKK significantly reduced UVB-induced IL1A gene expression relative to zinc or retinol. HaCaT keratinocytes were irradiated with 10 mJ/cm ² UVB and treated with FWAHKK, Zn-FWAHKK, ZnSO4*7H2O (Zn) and retinol for 24 h. The relative gene expression of IL-Ια (IL1A gene) was determined by qRT-PCR. *** p<0.001 compared to respective UVB controls.

Giant. 6. Hexapeptides FWAHKK and Zn-FWAHKK reduced the expression of genes associated with keratinocyte differentiation similar to retinol, while zinc had no such effect. HaCaT keratinocytes were treated with FWAHKK, Zn-FWAHKK, ZnSO4*7H2O (Zn) and retinol for 72 h. The relative gene expression of 5n-reductase (SRD5A1 gene) was determined by qRT-PCR * p<0.05; ** p<0.01; *** p<0.001 compared to respective controls.

Giant. 7. Hexapeptides FWAHKK and Zn-FWAHKK significantly reduced UVB-induced gene expression of IL6 and IL8 compared to zinc or retinol. HaCaT keratinocytes were irradiated with 10 mJ/cm ² UVB and treated with FWAHKK, Zn-FWAHKK, ZnSO4*7H2O (Zn) and retinol for 24 h. The relative gene expression of pro-inflammatory interleukins IL-6 and IL-8 (IL6 and IL8 genes) was determined by qRT-PCR. *p<0.05; ** p<0.01; *** p<0.001 compared to respective UVB controls.

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Giant. 8. Hexapeptide FWAHKK and zinc had antimicrobial activity against C. acnes growing in suspension. Zn-FWAHKK had higher activity than the individual components. C. acnes growing in suspension were cultured with FWAHKK, Zn-FWAHKK and ZnSO4*7H2O (Zn) for 72 h. Then OD590 was measured. *p<0.05; ** p<0.01; *** p<0.001 compared to control unless otherwise stated.

Giant. 9. Hexapeptides FWAHKK and Zn-FWAHKK stimulated collagen 1 gene expression compared to zinc or retinol. HaCaT keratinocytes were treated with FAWHKK, ZnFWAHKK, ZnSO4*7H2O (Zn), and retinol for 72 h. Relative gene expression of 5α-reductase (SRD5A1 gene) was determined by qRT-PCR. *p<0.05; ** p<0.01; *** p<0.001 compared to respective controls.

Giant. 10. The Zn-FWAHKK complex had anti-acne and anti-aging activity and is more effective than retinol. In vivo study of 40 volunteers with acne-prone skin who applied two emulsions with 13.5 pg/ml Zn-FWAHKK and placebo (30 subjects) or 0.2% retinol and placebo (10 subjects) on two halves of the face in given order once a day for 6 weeks. The number of acne lesions (A) was determined by image analysis of full-face images acquired by VisiaCR. The depth of crow's feet wrinkles (B) was determined by a 3D camera. *p<0.05; ** p<0.01, *** p<0.001 compared to placebo.

Examples of implementation of the invention

Example 1. Preparation of FWAHKK, Zn-FWAHKK, acetyl-FWAHKK and palmitoyl-FWAHKK peptide

First, the FWAHKK peptide was synthesized by standard solid-phase peptide synthesis using the Fmoc/tBu strategy on Wang resin (Amblard et al. 2006).

Attachment of the first amino acid to Wang resin (Sunresin, China) was performed by syringe reactor in dimethylformamide (DMF) using diisopropylcarbodiimide (DIC) and OxymaPure® (Iris Biotech, Germany) in 3 molar excess (meq) of Fmoc-Lys(Boc)-OH via night. After coupling, the residual hydroxyl groups on the resin were covered with a solution of acetic anhydride/pyridine (1/1, v/v).

The FWAHKK peptide was synthesized on a 0.1 mmol scale using a CEM Liberty Blue automated microwave peptide synthesizer (CEM, USA) on Wang resin saturated with first Fmoc-AA. Fmoc deprotections were performed with 20% pyridine in DMF. Coupling reactions were performed using 5 equivalents of Fmoc-AA/DIC/Oxyma Pure ^® in DMF.

To prepare acetyl-FWAHKK and palmitoyl-FWAHKK, the N-terminus of the synthesized FWAHKK peptide was acetylated or palmitoylated on the resin using 2 meq of acetaldehyde or palmitoyl anhydride/pyridine (1/1, v/v) in DMF for 30 min at room temperature.

Cleavage of peptides (FWAHKK, acetyl-FWAHKK or palmitoyl-FWAHKK) from the resin was performed by raising the temperature (38 °C) for 30 min using a CEM Razor® robust peptide cleavage system (CEM, USA) with a mixture of trifluoroacetic acid/anisole/thioanisole/H2O/ dichloromethane (85/2.5/2.5/5/5). The peptides were then precipitated in cold diethyl ether.

Crude FWAHKK and acetyl-FWAHKK peptides were dissolved in 50% acetonitrile, crude palmitoyl-FWAHKK was dissolved in 100% acetonitrile. They were then purified on Shimadzu preparative HPLC (Prominence series). XBridge OBD Prep Column Reversed-Phase 5 pm Spherical Hybrid, 50 mm x 50 mm was used for separation at a flow rate of 30 ml/min. The mobile phase was A 0.1% HCOOH in water and B acetonitrile. The purity of the peptides was verified by PDA and MS detector. The purified peptides were then lyophilized.

- 9 CZ 309857 B6

For the preparation of FWAHKK hexapeptide in complex with zinc (Zn-FWAHKK), an aqueous solution of 10% FWAHKK hexapeptide and 3.52% ZnSO4*7H2O (zinc sulfate, heptahydrate, Lachner, Czech Republic) corresponding to a molar ratio of 1:1 was prepared, stirred overnight at room temperature and then the solution was lyophilized.

Example 2. Ability of the FWAHKK peptide to form a complex with zinc

The ability of the FWAHKK peptide to form a complex with zinc was evaluated as previously described with minor modifications (Catapano et al., 2018). Briefly, 20 µl of FWAHKK solution (0 to 600 μΜ (0 to 490 pg/ml; 0 to 0.049 w/w%) prepared as described in Example 1) and 60 μΜ (17.3 pg/ml; 0 .00173%) ZnSO4*7H2O in 15 mM HEPES, pH 6.8 was mixed with 20 μl of 250 μΜ dithizone in DMSO and 80 μl of 15 mM HEPES, pH 6.8. Dithizone was used as an indicator of free zinc. After 30 min of incubation at room temperature (RT), the absorbance at 520 nm was measured and the percentage of zinc complexed with the FWAHKK peptide was calculated.

The results (Fig. 1) confirmed the ability of the FWAHKK peptide to complex with zinc. In the Zn-FWAHKK complex prepared as described in Example 1, in which the molar ratio of FWAHKK to zinc is 1:1, approximately 50% of the zinc is complexed with the hexapeptide.

Example 3. Increased penetration of zinc in the Zn-FWAHKK complex into cells

HaCaT keratinocytes (CLS collection, Germany) were cultured in Dubelc's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum, 0.2 mg/ml glutamine, 100 U/ml penicillin, and 0.1 mg/ml streptomycin (all Sigma-Aldrich). Aldrich, USA) at 37 °C in a humidified atmosphere with 5% CO2.

Cells seeded at the appropriate density in 96-well plates were treated with 1352 pg/ml (0.1352%) Zn-FWAHKK complex (prepared as described in Example 1) or ZnSO4*7H2O at the corresponding concentration (352 pg/ml (0 .0352 %)) for 3 h. Then the cells were washed with cell culture medium and incubated with 1 pM FluoZin ^TM -3 AM (Invitrogen, USA), a cell permeation agent, a fluorogenic Zn ²⁺ -selective indicator, and 20 pM Hoechst (Abcam, UK), fluorescent DNA dye in cell culture medium for 30 min at 37 °C in the dark. The cells were then rinsed with phosphate-buffered saline (PBS). Fluorescence was observed using an Eclipse 50i fluorescence microscope equipped with a DS-Fi1 camera (both Nikon, Japan). The results (Fig. 2) showed that zinc penetration into skin cells was more efficient when the ion was in the Zn-FWAHKK complex.

Example 4. FWAHKK increases cell viability and prevents the cytotoxic effect of zinc in the Zn-FWAHKK complex

NIH-3T3 mouse embryonic fibroblasts (ATCC collection, USA) were cultured in the same manner as HaCaT keratinocytes as described in Example 3. Cells seeded at an appropriate density in 96-well plates were treated with FWAHKK (2.5 to 80 pg/ml ( 0.00025 to 0.008%)), Zn-FWAHKK (3.4 to 108 pg/ml (0.00034 to 0.0108%)) (both prepared as described in Example 1), and ZnSO4*7H2O (0. 9 to 28.2 pg/ml (0.00009 to 0.00282%)) 48 h. Then cell viability was determined by the MTT method (Riss et al., 2016). Briefly, cells were incubated with 0.5 mg/ml MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) in cell culture medium for 2.5 h at 37°C. After MTT removal, cell lysis and formazan dissolution was performed by incubating cells with solubilization solution (45% isopropanol, 45% DMSO, 10% Triton-X, 0.3 M HCl) for 30 min at RT, shaking. The absorbance was then measured at 570 nm with a spectrophotometer (EnVision® 2105 Multimode Plate Reader, Perkin, Elmer, USA). T-test was used for statistical evaluation of the results.

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The results (Fig. 3) showed that ZnSO4*7H2O in concentrations from 10.6 μg/ml significantly reduced cell viability. On the other hand, the FWAHKK peptide increased cell viability and the formation of the ZnFWAHKK complex prevented the cytotoxic effect of zinc.

Example 5. Inhibition of 5α-reductase gene expression of FWAHKK and Zn-FWAHKK

HaCaT keratinocytes (CLS collection, Germany) were cultured as described in Example 3. Cells seeded at an appropriate density in 6-well plates were treated with FWAHKK (1 μg/ml (0.0001%) and 10 μg/ml (0.001% )), Zn-FWAHKK (1.35 μg/ml (0.000135%) and 13.5 μg/ml (0.00135%)) (both prepared as described in Example 1), ZnSO4*7H2O (Zn , 0.35 μg/ml (0.000035%) and 3.5 μg/ml (0.00035%)), and 10 μΜ retinol (in DMSO, final concentration of DMSO was 0.1%) for 72 h. Control cells were untreated or treated with 0.1% DMSO in case of retinol control.

Then, the expression of the 5α-reductase gene (SRD5A1 gene) was quantitatively determined by real-time reverse transcription PCR (qRT-PCR) as follows. Total RNA was isolated from cells by the guanidine thiocyanate-phenol acid extraction method using TRI Reagent (Sigma Aldrich, USA) according to the instructions provided by the supplier. Reverse transcription was performed using the Large Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific, MA, USA) in a GenePro thermal cycler (Bioer Technology, China) according to the manufacturer's instructions. Subsequent qPCR was performed by a specific TaqMan gene expression method for SRD5A1 (qHsaCEP0052536, BioRad, CA, USA), and RPL13A (Hs04194366_g1, Thermo Fisher Scientific, MA, USA) as a reference gene; and TaqMan Fast Advanced Master mix (all Thermo Fisher Scientific, MA, USA) as recommended by the supplier in a One-Step Real-time PCR cycler (Thermo Fisher Scientific, MA, USA). Data were analyzed using the 2 ^-ΔΔα method. Data were normalized to negative controls or, in the case of retinol, to DMSO controls. The ttest was used for statistical evaluation of the data.

5α-reductase is a key enzyme involved in androgen-induced sebum production. This enzyme converts testosterone, the most common androgen, to DHT, which is several times more effective at stimulating sebum production. The results (Fig. 4) showed that the hexapeptide FWAHKK, ZnFWAHKK and also ZnSO4*7H2O significantly decreased the expression of the 5a-reductase gene (SRD5A1 gene). The Zn-FWAHKK complex was more effective than its individual components at the corresponding concentrations. On the other hand, retinol had no significant effect on 5αreductase gene expression.

Example 6. IL-1α inhibition of FWAHKK and Zn-FWAHKK gene expression

HaCaT keratinocytes (CLS collection, Germany) were cultured as described in Example 3. Cells seeded at the appropriate density in 6-well plates were washed in PBS and irradiated with 10 mJ/cm ² UVB using a 300 W xenon arc lamp (Oriel Arc Lamp Housing, Newport, CA, USA) and a 300±10 nm optical filter (Newport, CA, USA). They were then treated with FWAHKK (1μg/ml (0.0001%) and 10μg/ml (0.001%), Zn-FWAHKK (1.35μg/ml (0.0001%) and 13.5μg/ml (0.001 %)) (both prepared as described in Example 1), ZnSO4*7 H2O (Zn; 0.35 μg/ml (0.000035%) and 3.5 μg/ml (0.00035%)) and 10 μM retinol (in DMSO, final DMSO concentration was 0.1%) 24 h. Control cells were untreated or treated with 0.1% DMSO in case of retinol control.

IL1A gene expression was determined by real-time qRT-PCR as described in Example 5, using a specific TaqMan gene expression method for IL1A (qHsaCIP0030471, BioRad, CA, USA), and RPL13A (Hs04194366_g1, Thermo Fisher Scientific, MA, USA) as a reference gene. Data were analyzed using the 2 ^-ΔΔCt method. Data were normalized to UVB untreated controls or, in the case of retinol, to UVB DMSO controls. T-test was used for statistical evaluation of the results.

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IL-1α is a pro-inflammatory interleukin stimulating hyperkeratinization of follicular keratinocytes (Guy and Kealey 1998) in the early stages of acne pathogenesis. In our experiments, we induced IL-1α gene expression in HaCaT keratinocytes using UVB irradiation (Fig. 5). Then, we observed a significant inhibition of UVB-induced IL1A gene expression after FWAHKKa Zn-FWAHKK treatment, while zinc and retinol showed no effect (Fig. 5).

Example 7. Inhibition of keratinocyte differentiation by FWAHKK and Zn-FWAHKK

HaCaT keratinocytes (CLS collection, Germany) were cultured as described in Example 3. Cells seeded at an appropriate density in 6-well plates were treated with FWAHKK (1 μg/ml (0.0001%) and 10 μg/ml (0.001%) )), Zn-FWAHKK (1.35 μg/ml (0.000135%) and 13.5 μg/ml (0.00135%)) (both prepared as described in Example 1), ZnSO4*7H2O (Zn; 0.35 μg/ml (0.000035%) and 3.5 μg/ml (0.00035%)) and 10 μΜ retinol (in DMSO, final DMSO concentration was 0.1%) for 72 h.

Expression of selected genes involved in keratinocyte differentiation was determined by real-time qRTPCR as described in Example 5 using the specific TaqMan gene expression method for FLG (Hs00856927_g1), OCLN (Hs00170162_m1), LCE2C (Hs02390636_s1), SPRR2E (qHsaCEP0055677, BioRad, CA , USA), KRT10 (Hs01043114_g1) and RPL13A (Hs04194366_g1) as a reference gene (all except SPRR2E Thermo Fisher Scientific, MA, USA). Data were analyzed using the 2 ^-AACt method. Data were normalized to untreated controls or, in the case of retinol, to DMSO controls. T-test was used for statistical evaluation of the results.

Hexapeptide FWAHKK as well as Zn-FWAHKK were shown to downregulate genes associated with keratinocyte differentiation in the same manner as 10 μΜ retinol, whereas zinc at the corresponding concentrations had no such effect (Fig. 6).

Example 8. Anti-inflammatory effect of FWAHKK and Zn-FWAHKK

HaCaT keratinocytes (CLS collection, Germany) were cultured as described in Example 3. Cells seeded at the appropriate density in 6-well plates were washed in PBS and irradiated with 10 mJ/cm ² UVB using a 300 W xenon arc lamp (Oriel Arc Lamp Housing, Newport, CA, USA) and a 300 ±10 nm optical filter (Newport, CA, USA). Then they were treated with FWAHKK (1 μg/ml (0.0001%) and 10 μg/ml (0.001%), Zn-FWAHKK (1.35 μg/ml (0.000135%) and 13.5 μg/ml (0. 00135%)) (both prepared as described in Example 1), ZnSO4*7H2O (Zn, 0.35 μg/ml (0.000035%) and 3.5 μg/ml (0.00035%)), and 10 μΜ of retinol (in DMSO, final DMSO concentration was 0.1%) for 24 h. Control cells were untreated or treated with 0.1% DMSO in the case of retinol control.

IL1A gene expression was determined by real-time qRT-PCR as described in Example 5 using the specific TaqMan gene expression method for IL6 (Hs00174131_m1), IL8 (Hs00174103_m1), and RPL13A (Hs04194366_g1) as a reference gene (all Thermo Fisher Scientific, MA, USA). Data were analyzed using the 2 ^-AACt method. Data were normalized to UVB untreated controls or, in the case of retinol, to UVB DMSO controls. T-test was used for statistical evaluation of the results.

Inflammation is a key event in the pathogenesis of acne. We induced inflammatory processes as increased expression of pro-inflammatory interleukins IL-6 and IL-8 in HaCaT keratinocytes using UVB radiation (Fig. 7.) We observed a significant inhibition of UVB-induced IL6 and IL8 gene expression after FWAHKK and Zn-FWAHKK treatment similar to 10 μM retinol, while zinc at the corresponding concentrations showed no effect (Fig. 7).

Example 9. Antimicrobial activity of FWAHKK and Zn-FWAHKK against C. acnes

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Cutibacterium acnes (strain DSM 1897, DSZM collection, Germany) was maintained in suspension in tryptic soy broth (TSB) at 37°C. Bacteria were then inoculated at an appropriate density into TSB containing FWAHKK (10 to 80 μg/ml (0.001 to 0.008%)), Zn-FWAHKK (13.5 to 108.2 μg/ml (0.00135 to 0.01082%) ) (both prepared as described in Example 1) and ZnSO4*7H2O (Zn; 3.5 to 28.2 μg/ml (0.00035 to 0.00282%)) in 96-well plates. The bacteria were cultured for 72 h at 37 °C and then the optical density at 590 nm (OD590) was measured with a spectrophotometer (EnVision® 2105 Multimodal plate reader, PerkinElmer, USA). T-test was used for statistical evaluation of the results.

Both zinc and the hexapeptide FWAHKK had weak antimicrobial activity against C. acnes (Fig. 8). The Zn-FWAHKK complex was more effective than the individual compounds at the corresponding concentrations (Fig. 8).

Example 10. Stimulation of collagen production by FWAHKK and Zn-FWAHKK

NIH-3T3 mouse embryonic fibroblasts (ATCC collection, USA) were cultured in the same manner as HaCaT keratinocytes as described in Example 3. Cells seeded at an appropriate density in 6-well plates were treated with FWAHKK (1 μg/ml (0.0001% ) and 10 pg/ml (0.001%)), Zn-FWAHKK (1.35 pg/ml (0.000135%) and 13.5 pg/ml (0.00135%)) (both prepared as described in Example 1 ), ZnSO4*7H2O (Zn, 0.35 pg/ml (0.000035%) and 3.5 pg/ml (0.00035%)), and 10 μΜ retinol (in DMSO, final DMSO concentration was 0.1 %) 72 h. Control cells were untreated or treated with 0.1% DMSO in case of retinol control.

COL1A1 gene expression was determined by real-time qRT-PCR as described in Example 5, using the specific TaqMan gene expression method for COL1A1 (Mm00801666_g1), and RPL13A (Mm05910660_g1) as a reference gene (both Thermo Fisher Scientific, MA, USA). Data were analyzed using ^AACt method 2. Data were normalized to untreated controls or, in the case of retinol, to DMSO controls. T-test was used for statistical evaluation of the results.

The results showed a significant stimulation of collagen 1 gene expression after treatment with FWAHKK and ZnFWAHKK, while zinc at the corresponding concentrations and 10 μΜ retinol showed no effect (Fig. 9).

Example 11 Zn-FWAHKK improves the appearance of acne-prone skin and also has an antiaging effect in vivo

We conducted a double-blind, placebo-controlled, in vivo face-splitting study in 40 human volunteers (Caucasian, Fitzpatrick skin type I-III, female/male 36/4, 18 to 49 years, mean 30.5 years) with oily, problematic skin prone to acne. This study was conducted in accordance with the principles of the Declaration of Helsinki of the World Medical Association. This study was approved by the ethical committee of Contipro a.s. and informed consent was obtained from all volunteers. Only healthy subjects without acute facial skin disorders were included in the study. Other exclusive criteria were pregnancy, breastfeeding, invasive cosmetic facial procedures (face lift, botox, etc.) and heavy smoking (> 10 cigarettes per day). Application of any facial skin care products as well as face washing was not allowed during the measurement day.

All volunteers received two emulsions: 30 volunteers (female/male 27/3, 18 to 48 years, mean 31.0 years) received an emulsion with 0.00135% Zn-FWAHKK (13.5 μg/ml; prepared as described in Example 1) and placebo, and 10 volunteers (female/male 9/1, 24 to 49 years, mean 28.9 years) received 0.2% retinol emulsions and placebo. The composition of the emulsions is described in Table 1. Two emulsions were applied to two individual halves of the face once a day in the evening for 6 weeks. Skin parameters were measured at the beginning of the study and then after 2, 4 and 6 weeks.

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The measurements were performed after 30 min of acclimatization of the volunteers in a room with controlled conditions (T: 20 to 22 °C; RH 40 to 45%). The number of acne lesions was determined by image analysis of full-face images acquired by VisiaCR, a high-resolution camera (Canfield Scientific, USA). Quantification was performed using Image-Pro 10 analysis software (Media Cybernetics, USA). Crow's feet wrinkle depth was determined using a Primos Lite 3D camera (Canfield Scientific, USA).

The results showed an improvement in the appearance of acne-prone skin represented by a reduction in the number of acne lesions (Fig. 10A). Zn-FWAHKK was also shown to have antiaging activity by reducing wrinkles (Fig. 10B). In both cases, Zn-FWAHKK was more effective than 0.2% retinol.

Table 1 Composition of the emulsions used in the in vivo study

INCI (name) % w/w Placebo Zn-FWAHKK Retinol Aqua 96.52 96.51865 96.32 Capric/Caprylic Triglycerides 1.00 1.00 1.00 Phenoxyethanol 1.00 1.00 1.00 Hydroxyethyl cellulose 0.70 0.70 0.70 Tocopheryl Acetate (Vitamin E) 0.50 0.50 0.50 Carbomer 0.25 0.25 0.25 Sodium Hydroxide 0.03 0.03 0.03 Retinol - - 0.20 Zn-FWAHKK 0.00135

Example 12. Composition of emulsions with FWAHKK, Zn-FWAHKK, acetyl-FWAHKK or palmitoylFWAHKK

Example of creams (oil-in-water emulsion) containing hexapeptides of the present invention FWAHKK, Zn-FWAHKK, acetyl-FWAHKK or palmitoyl-FWAHKK prepared as described in Example 1.

Table 2.

Component INCI Cream with: (% w/w) FWAHKK ZnFWAHKK AcetylFWAHKK PalmitoylFWAHKK Oil phase Emulsifying mixture Glyceryl Stearate, PEG-100 Stearate 5.00 5.00 5.00 5.00 Macadamia oil Macadamia Integrifolia Seed Oil 5.00 5.00 5.00 5.00 Olive oil Olea europaea Fruit Oil 4.00 4.00 4.00 3.00 Jojoba oil Simmondsia Chinensis Seed Oil 3.00 3.00 3.00 3.00 Vitamin E Vitamin E Tocopheryl Acetate 1.00 1.00 1.00 1.00 Aqueous phase Water Aqua 65.50 65.50 65.50 70,70 Glycerine Glycerine 4.00 4.00 4.00 4.00 Gel maker EMU Sodium Acrylate / Sodium 2.00 2.00 2.00 2.00

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Acryloyldimethyl Taurate Copolymer, Isohexadecane, Polysorbate 80 Urea Urea 3.00 3.00 3.00 3.00 Vitamin B3 Niacinamide 1.50 1.50 1.50 1.50 Phases of water-soluble active ingredients Water Aqua 5,1999 5.199865 5,1999 - FWAHKK - 0.0001 - - - ZnFWAHKK - - 0.000135 - - AcetylFWAHKK - - - 0.0001 - Phase of oil-soluble active ingredients Olive oil Olea Europaea Fruit Oil - - - 0.9999 PalmitoylFWAHKK - - - - 0.0001 Preservative Benzyl alcohol-DHA Benzyl Alcohol, Dehydroacetic Acid 0.80 0.80 0.80 0.80

Table 3

Component INCI Cream with: (% w/w) FWAHKK ZnFWAHKK AcetylFWAHKK PalmitoylFWAHKK Oil phase Lamecreme Glyceryl Stearate, Glyceryl Stearate Citrate 6.00 6.00 6.00 6.00 Macadamia oil Macadamia Integrifolia Seed Oil 6.00 6.00 6.00 6.00 Olive oil Olea Europaea Fruit Oil 16.00 16.00 16.00 15.00 Jojoba oil Simmondsia Chinensis Seed Oil 8.00 8.00 8.00 8.00 Cetyl alcohol Cetyl Alcohol 2.00 2.00 2.00 2.00 Aqueous phase Water Aqua 47.30 47.30 47.30 47.30 Glycerine Glycerine 4.00 4.00 4.00 4.00 Urea Urea 3.00 3.00 3.00 3.00 Vitamin B3 Niacinamide 1.50 1.50 1.50 1.50 Phases of water-soluble active ingredients Water Aqua 5,189 5.18865 5,189 5.19 HyRetin Sodium Retinoyl Hyaluronate 0.01 0.01 0.01 0.01 FWAHKK - 0.001 - - - ZnFWAHKK - - 0.00135 - -

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AcetylFWAHKK - - - 0.001 - Phase of oil-soluble active ingredients Olive oil Olea Europaea Fruit Oil - - - 0.999 PalmitoylFWAHKK - - - - 0.001 Preservative Phenoxy ethane ol Phenoxyethanol 1.00 1.00 1.00 1.00

Table 4

Component INCI Cream with: (% w/w FWAHKK ZnFWAHKK AcetylFWAHKK PalmitoylFWAHKK Oil phase Cetearyl alcohol Cetearyl Alcohol 6.00 6.00 6.00 6.00 Macadamia oil Macadamia Integrifolia Seed Oil 6.00 6.00 6.00 6.00 am Olive oil Olea Europaea Fruit Oil 11.00 11.00 11.00 10.00 Jojoba oil Simmondsia Chinensis Seed Oil 8.00 8.00 8.00 8.00 Glyceryl stearate SE Glyceryl Stearate SE 2.00 2.00 2.00 2.00 Aqueous phase Water Aqua 52.30 52.30 52.30 57.50 Glycerine Glycerine 4.00 4.00 4.00 4.00 Urea Urea 3.00 3.00 3.00 3.00 Vitamin B3 Niacinamide 1.50 1.50 1.50 1.50 Phases of water-soluble active ingredients Water Aqua 5.19 5.1865 5.19 - FWAHKK - 0.01 - - - Zn-FWAHKK - - 0.0135 - - AcetylFWAHKK - - - 0.01 - Phase of oil-soluble active ingredients Olive oil Olea Europaea Fruit Oil - - - 0.99 PalmitoylFWAHKK - - - - 0.01 Preservative Phenoxyethanol Phenoxyethanol 1.00 1.00 1.00 1.00

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Table 5

Component INCI Cream with: (% w/w FWAHKK ZnFWAHKK AcetylFWAHKK PalmitoylFWAHKK Oil phase Almond oil Prunus Amygdalus Dulcis Oil 15.00 15.00 15.00 14.00 Shea Butter Butyrospermum Parkii Butter 5.00 5.00 5.00 5.00 Cetyl alcohol Cetyl Alcohol 2.00 2.00 2.00 2.00 Glyceryl Stearate Citrate Glyceryl Stearate Citrate 5.00 5.00 5.00 5.00 Aqueous phase Water Aqua 50.90 50.90 50.90 50.90 Phases of water-soluble active ingredients Water Aqua 10.89 10,855 10.89 10.99 Glycerine Glycerine 4.00 4.00 4.00 4.00 Panthenol Panthenol 2.00 2.00 2.00 2.00 TanActin Glucomannan 0.20 0.20 0.20 0.20 Schizophyllan Schizophyllan 0.10 0.10 0.10 0.10 Urea Urea 4.00 4.00 4.00 4.00 HyRetin Sodium Retinoyl Hyaluronate 0.01 0.01 0.01 0.01 FWAHKK - 0.10 - - - Zn-FWAHKK - - 0.135 - - AcetylFWAHKK - - - 0.10 - Phase of oil-soluble active ingredients Almond oil Prunus Amygdalus Dulcis Oil - - - 0.90 PalmitoylFWAHKK - - - - 0.10 Preservative Benzyl alcohol-DHA Benzyl Alcohol, Dehydroacetic Acid 0.80 0.80 0.80 0.80

The oil and water phases were prepared and heated to 70°C. Next, both phases were mixed and emulsified under stirring (290 rpm/min). The emulsion was then cooled to 40°C and a phase of water-soluble active ingredients and/or vitamin E and a preservative was added. Finally, the pH was measured and adjusted to values between 2.2 and 6.5 1 to. 50% citric acid or 1 to 50% KOH or NaOH.

O

Example 13. Composition of sera with FWAHKK, Zn-FWAHKK and acetyl-FWAHKK

Example of sera containing hexapeptides of the present invention FWAHKK and Zn-FWAHKK prepared as described in Example 1.

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Table 6

Component INCI Serum with: (% w/w) FWAHKK ZnFWAHKK AcetylFWAHKK Water Aqua 97.7999 97.799865 97.7999 Carbopol Ultrez 10 Carbomer 0.60 0.60 0.60 30% Sodium hydroxide Aqua, Sodium Hydroxide 0.50 0.50 0.50 Sodium hyaluronate Sodium Hyaluronate 0.10 0.10 0.10 FWAHKK - 0.0001 - - Zn-FWAHKK - - 0.000135 - Acetyl-FWAHKK - - - 0.0001 Pheno xy ethanol Phenoxyethanol 1.00 1.00 1.00

Table 7

Component INCI Serum with: (% w/w) FWAHKK ZnFWAHKK AcetylFWAHKK Water Aqua 94,799 94.79865 94,799 Sodium hyaluronate Sodium Hyaluronate 1.50 1.50 1.50 Vitamin B3 Niacinamide 1.50 1.50 1.50 TanActin Glucomannan 0.20 0.20 0.20 FWAHKK - 0.001 - - Zn-FWAHKK - - 0.00135 - Acetyl-FWAHKK - - - 0.001 Phenoxy ethanol Phenoxyethanol 1.00 1.00 1.00

Table 8

Component INCI Serum with: (% w/w) FWAHKK ZnFWAHKK AcetylFWAHKK Water Aqua 95.69 90.6865 95.69 Ecogel Lysolecithin, Sclerotium Gum, Xanthan Gum, Pullulan 1.50 1.50 1.50 Sodium hyaluronate Sodium Hyaluronate 0.10 0.10 0.10 Vitamin B3 Niacinamide 1.50 1.50 1.50 Vitamin C Sodium Ascorbyl Phosphate 5.00 5.00 5.00 TanActin Glucomannan 0.20 0.20 0.20 FWAHKK - 0.01 - - Zn-FWAHKK - - 0.0135 - Acetyl-FWAHKK - - - 0.01 Phenoxy ethanol Phenoxyethanol 1.00 1.00 1.00

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Table 9

Component INCI Serum with: (% wt/ property) FWAHKK ZnFWAHKK AcetylFWAHKK Water Aqua 92.20 92,165 92.20 Crosslinked ^HA Sodium Hyaluronate and Sodium Hyaluronate Crosspolymer-3 1.50 1.50 1.50 Sodium hyaluronate Sodium Hyaluronate 0.90 0.90 0.90 Vitamin B3 Niacinamide 1.50 1.50 1.50 Panthenol Panthenol 1.00 1.00 1.00 Vitamin C Sodium Ascorbyl Phosphate 1.80 1.80 1.80 TanActin Glucomannan 0.20 0.20 0.20 FWAHKK - 0.1 - - Zn-FWAHKK - - 0.135 - Acetyl-FWAHKK - - - 0.1 Benzyl alcoholDHA Benzyl Alcohol, Dehydroacetic Acid 0.80 0.80 0.80

First, hexapeptide FWAHKK or Zn-FWAHKK or acetyl-FWAHKK and active ingredients except sodium hyaluronate and croslink ^HA were added to water and stirred until completely dissolved. Next, thickeners and/or sodium hyaluronate and croslink ^HA were added and the mixture was mixed until the components were completely dissolved. A preservative was then added. Finally, the pH was measured and adjusted to values between 5.2 and 6.5 with 1 to 50% citric acid or 1 to 50% KOH or NaOH.

Literature review

Abendrot, M., and U. Kalinowska-Lis. 2018. “Zinc-Containing Compounds for Personal Care Applications.” International Journal of Cosmetic Science 40 (4): 319-27. https://doi.Org/10.l 111/ics.12463.

Amblard, Muriel, Jean-Alain Fehrentz, Jean Martinez, and Gilles Subra. 2006. “Methods and Protocols of Modern Solid Phase Peptide Synthesis.” Molecular Biotechnology 33 (3): 23954. https://doi.Org/10.1385/MB:33:3:239.

Anderson, K.M., and S. Liao. 1968. “Selective Retention of Dihydrotestosterone by Prostatic Nuclei.” Nature 219 (5151): 277-79. https://doi.org/10.1038/219277a0.

Arif, Tasleem. 2015. “Salicylic Acid as a Peeling Agent: A Comprehensive Review.” Clinical, Cosmetic and Investigational Dermatology 8 (August): 455-61.

https://doi.org/10.2147/CCID.S84765.

Bowe, Whitney P., Jennifer C. Filip, Joseph M. DiRienzo, Alla Volgina, and David J. Margolis. 2006. "Inhibition of Propionibacterium Acnes by Bacteriocin-like Inhibitory Substances (BLIS) Produced by Streptococcus Salivarius." Journal of Drugs in Dermatology: JDD 5 (9): 868-70.

Catapano, Maria Carmen, Václav Tvrdý, Jana Karlíčková, Laura Mercolini, and Přemysl Mladěnka. 2018. “A Simple, Cheap but Reliable Method for Evaluation of Zinc Chelating Properties.” Bioorganic Chemistry ΊΊ (April): 287-92.

https://doi.Org/10.1016J.bioorg.2018.01.015.

Cebrián, Rubén, Sergio Arévalo, Susana Rubino, Salvador Arias-Santiago, Maria Dolores Rojo, Manuel Montalbán-López, Manuel Martinez-Bueno, Eva Valdivia, and Mercedes Maqueda. 2018. “Control of Propionibacterium Acnes by Natural Antimicrobial Substances: Role of

- 19CZ 309857 B6 the Bacteriocin AS-48 and Lysozyme.” Scientific Reports 8 (1): 11766.

https://doi.org/10.1038/s41598-018-29580-7.

Cervantes, Jessica, Ariel E. Eber, Marina Perper, Vanessa M. Nascimento, Keyvan Nouri, and Jonette E. Keri. 2018. “The Role of Zinc in the Treatment of Acne: A Review of the Literature.” Dermatologic Therapy 31 (1). https://doi.org/10.1111/dth.12576.

Cunha, Marisa Gonzaga da, Fernando Luiz Affonso Fonseca, and Carlos D. Aparecida S. Machado. 2013. “Androgenic Hormone Profile of Adult Women with Acne.” Dermatology (Basel, Switzerland) 226(2): 167-71. https://doi.org/10.1159/000347196.

Downing, D.T., M.E. Stewart, P.W. Wertz, and J.S. Strauss. 1986. “Essential Fatty Acids and Acne.” Journal of the American Academy of Dermatology 14(2 Pt 1): 221-25. https://doi.org/10.1016/s0190-9622(86)70025-x.

Dréno, B., S. Pécastaings, S. Corvec, S. Veraldi, A. Khammari, and C. Roques. 2018. “Cutibacterium Acnes (Propionibacterium Acnes) and Acne Vulgaris: A Brief Look at the Latest Updates.” Journal of the European Academy of Dermatology and Venereology 32 (S2): 5-14. https://doi.org/10.1111/jdv.15043.

Fritsch, M., C.E. Orfanos, and C.C. Zouboulis. 2001. “Sebocytes Are the Key Regulators of Androgen Homeostasis in Human Skin.” The Journal of Investigative Dermatology 116 (5): 793-800. https://doi.org/10.1046ij.1523-1747.2001.01312.x.

Guy, R., and T. Kealey. 1998. "The Effects of Inflammatory Cytokines on the Isolated Human Sebaceous Infundibulum." The Journal of Investigative Dermatology 110 (4): 410-15. https://doi.org/10.1046/j.1523-1747.1998.00143.x.

Han, Rui, Hans-Matti Blencke, Hao Cheng, and Chun Li. 2018. “The Antimicrobial Effect of CEN1HC-Br against Propionibacterium Acnes and Its Therapeutic and Anti-Inflammatory Effects on Acne Vulgaris.” Peptides 99 (January): 36-43.

https://doi.org/10.1016/j.peptides.2017.11.001.

Kang, Bong Seon, Jae-Gu Seo, Gwa-Su Lee, Jung-Hwa Kim, Sei Yeon Kim, Ye Won Han, Hoon Kang, et al. 2009. “Antimicrobial Activity of Enterocins from Enterococcus Faecalis SL-5 against Propionibacterium Acnes, the Causative Agent in Acne Vulgaris, and Its Therapeutic Effect.” Journal of Microbiology (Seoul, Korea) 47(1): 101-9.

https://doi.org/10.1007/s12275-008-0179-y.

Kumtornrut, Chanat, Takeshi Yamauchi, Saaya Koike, Setsuya Aiba, and Kenshi Yamasaki. 2019. “Androgens Modulate Keratinocyte Differentiation Indirectly through Enhancing Growth Factor Production from Dermal Fibroblasts.” Journal of Dermatological Science 93 (3): 150-58. https://doi.org/10.1016/jjdermsci.2019.01.007.

Lim, Hee-Sun, Seung-Min Chun, Min-Gyu Soung, Jenny Kim, and Seong-Jin Kim. 2015. “Antimicrobial Efficacy of Granulysin-Derived Synthetic Peptides in Acne Vulgaris.” International Journal of Dermatology 54(7):853-62. https://doi.org/10.1111/ijd.12756.

Liu, Haibo, Haiyan Yu, Jun Xia, Ling Liu, Guan J. Liu, Hong Sang, and Frank Peinemann. 2020. “Topical Azelaic Acid, Salicylic Acid, Nicotinamide, Sulphur, Zinc and Fruit Acid (AlphaHydroxy Acid) for Acne.” The Cochrane Database of Systematic Reviews 5 (May):

Ma, Ziyuan, Nikolay Kochergin, Olga Olisova, and Elena Snarskaya. 2021. “Topical Antimicrobial Peptides in Combined Treatment of Acne Patients.” Journal of Cosmetic Dermatology, June. https://doi.org/10.1111/jocd.14300.

McInturff, Jamie E., Robert L. Modlin, and Jenny Kim. 2005. “The Role of Toll-like Receptors in the Pathogenesis and Treatment of Dermatological Disease.” Journal of Investigative Dermatology 125 (1): 1-8. https://doi.org/10.1111/j.0022-202X.2004.23459.x.

McLaughlin, Joseph, Steven Watterson, Alison M. Layton, Anthony J. Bjourson, Emma Barnard, and Andrew McDowell. 2019. “Propionibacterium Acnes and Acne Vulgaris: New Insights from the Integration of Population Genetic, Multi-Omic, Biochemical and Host-Microbe Studies.” Microorganisms 7 (5). https://doi.org/10.3390/microorganisms7050128.

Melo, Manuel N., Dominique Dugourd, and Miguel A.R.B. Castanho. 2006. “Omiganan Pentahydrochloride in the Front Line of Clinical Applications of Antimicrobial Peptides.” Recent Patents on Anti-Infective Drug Discovery 1 (2): 201-7.

https://doi.org/10.2174/157489106777452638.

- 20 CZ 309857 B6

Miazga-Karska, Malgorzata, Katarzyna Michalak, and Grazyna Ginalska. 2020. “Anti-Acne Action of Peptides Isolated from Burdock Root—Preliminary Studies and Pilot Testing.” Molecules (Basel, Switzerland) 25 (9). https://doi.org/10.3390/molecules25092027.

Mills, Otto H., Maressa C. Criscito, Todd E. Schlesinger, Robert Verdicchio, and Ernest Szoke. 2016. “Addressing Free Radical Oxidation in Acne Vulgaris.” The Journal of Clinical and Aesthetic Dermatology 9 (1): 25-30.

Moradi Tuchayi, Sara, Evgenia Makrantonaki, Ruta Ganceviciene, Clio Dessinioti, Steven R. Feldman, and Christos C. Zouboulis. 2015. “Acne Vulgaris.” Nature Reviews. Disease Primers 1 (September): 15029. https://doi.org/10.1038/nrdp.2015.29.

Nair, Sithara S., Olga Y. Zolotarskaya, Matthew J. Beckwith, Dennis E. Ohman, and Kenneth J. Wynne. 2017. “A Polycation Antimicrobial Peptide Mimic without Resistance Buildup against Propionibacterium Acnes.” Macromolecular Bioscience 17 (9).

https://doi.org/10.1002/mabi.201700090.

Pan, Chieh-Yu, Jyh-Yih Chen, Tai-Lang Lin, and Cheng-Hui Lin. 2009. "In Vitro Activities of Three Synthetic Peptides Derived from Epinecidin-1 and an Anti-Lipopolysaccharide Factor against Propionibacterium Acnes, Candida Albicans, and Trichomonas Vaginalis." Peptides 30(6): 1058-68. https://doi.org/10.1016/j.peptides.2009.02.006.

Panchaud, Alice, Chantal Csajka, Paul Merlob, Christof Schaefer, Maya Berlin, Marco De Santis, Thierry Vial, et al. 2012. “Pregnancy Outcome Following Exposure to Topical Retinoids: A Multicenter Prospective Study.” The Journal of Clinical Pharmacology 52 (12): 1844-51. https://doi.org/10.1177/0091270011429566.

Popovic, Suzana, Edit Urban, Miodrag Lukic, and J. Michael Conlon. 2012. “Peptides with Antimicrobial and Anti-Inflammatory Activities That Have Therapeutic Potential for Treatment of Acne Vulgaris.” Peptides 34 (2): 275-82.

https://doi.org/ 10.1016/j .peptides.2012.02.010.

Riss, Terry L., Richard A. Moravec, Andrew L. Niles, Sarah Duellman, Hélene A. Benink, Tracy J. Worzella, and Lisa Minor. 2016. Cell Viability Assays. Assay Guidance Manual [Internet]. Eli Lilly & Company and the National Center for Advancing Translational Sciences. https://www.ncbi.nlm.nih.gov/books/NBK144065/.

Rubinchik, Evelina, Dominique Dugourd, Teresa Algara, Christopher Pasetka, and H. David Friedland. 2009. “Antimicrobial and Antifungal Activities of a Novel Cationic Antimicrobial Peptide, Omiganan, in Experimental Skin Colonization Models.” International Journal of Antimicrobial Agents 34 (5): 457-61.

https://doi.org/10.1016/j.ijantimicag.2009.05.003.

Ryu, S., Y. Park, B. Kim, S.-M. Cho, J. Lee, H.-H. Lee, C. Gurley, et al. 2014. “Inhibitory and Anti-Inflammatory Effects of the Helicobacter Pylori-Derived Antimicrobial Peptide HPA3NT3 against Propionibacterium Acnes in the Skin.” The British Journal of Dermatology 171 (6): 1358-67. https://doi.org/10.1111/bjd.13480.

Ryu, Sunhyo, Hyo Mi Han, Peter I. Song, Cheryl A. Armstrong, and Yoonkyung Park. 2015. "Suppression of Propionibacterium Acnes Infection and the Associated Inflammatory Response by the Antimicrobial Peptide P5 in Mice." PloS One 10 (7): e0132619.

https://doi.org/10.1371/journal.pone.0132619.

Tanghetti, Emil A. 2013. “The Role of Inflammation in the Pathology of Acne.” The Journal of Clinical and Aesthetic Dermatology 6 (9): 27-35.

Thiboutot, D. M. 2000. “The Role of Follicular Hyperkeratinization in Acne.” Journal of Dermatological Treatment 11 (SUPPL. 2): 5-8.

https://doi.org/10.1080/095466300750163645.

Thielitz, Anja, and Harald Gollnick. 2008. “Topical Retinoids in Acne Vulgaris: Update on Efficacy and Safety.” American Journal of Clinical Dermatology 9 (6): 369-81.

https://doi.org/10.2165/0128071 -200809060-00003.

Walsh, Timothy R., John Efthimiou, and Brigitte Dréno. 2016. “Systematic Review of Antibiotic Resistance in Acne: An Increasing Topical and Oral Threat.” The Lancet Infectious Diseases 16 (3): e23-33. https://doi.org/10.1016/S1473-3099(15)00527-7.

Wang, Yipeng, Zhiye Zhang, Lingling Chen, Huijuan Guang, Zheng Li, Hailong Yang, Jianxu Li, Dewen You, Haining Yu, and Ren Lai. 2011. “Cathelicidin-BF, and Snake Cathelicidin

- 21 CZ 309857 B6

Derived Antimicrobial Peptide, Could Be an Excellent Therapeutic Agent for Acne Vulgaris.” PloS One 6 (7): e22120. https://doi.org/10.1371/joumal.pone.0022120.

Woodburn, Kathryn W, Jesse Jaynes, and L. Edward Clemens. 2020. “Designed Antimicrobial Peptides for Topical Treatment of Antibiotic Resistant Acne Vulgaris.” Antibiotics 9 (1): 23. https://doi.org/10.3390/antibiotics9010023.

Wu, Yun, Yuanyuan Qiang, Kun Cao, Wei Zhang, and Guangxian Zhang. 2020. “Inhibitory Effect of the Antimicrobial Peptide BLP-7 against Propionibacterium Acnes and Its AntiInflammatory Effect on Acne Vulgaris.” Toxicon: Official Journal of the International Society on Toxinology 184 (September): 109-15.

https://doi.org/10.1016/j.toxicon.2020.06.006.

Wu, Yun, Guangxian Zhang, and Maojun Zhou. 2020. “Inhibitory and Anti-Inflammatory Effects of Two Antimicrobial Peptides Moronecidin and Temporin-1Dra against Propionibacterium Acnes in Vitro and in Vivo.” Journal of Peptide Science: An Official Publication of the European Peptide Society 26 (7): e3255. https://doi.org/10.1002/psc.3255.

Yee, Brittany E., Phillip Richards, Jennifer Y. Sui, and Amanda Fleming Marsch. 2020. “Serum Zinc Levels and Efficacy of Zinc Treatment in Acne Vulgaris: A Systematic Review and Meta-Analysis.” Dermatologic Therapy 33(6):e14252. https://doi.org/10.1111/dth.14252.

Yentzer, Brad A., Jeff Hick, Erin L. Reese, Adam Uhas, Steven R. Feldman, and Rajesh Balkrishnan. 2010. “Acne Vulgaris in the United States: A Descriptive Epidemiology.” Cutis 86 (2): 94-99.

Zhang, Zhiye, Lixian Mu, Jing Tang, Zilei Duan, Fengyu Wang, Lin Wei, Mingqiang Rong, and Ren Lai. 2013. “A Small Peptide with Therapeutic Potential for Inflammatory Acne Vulgaris.” PloS One 8(8):e72923. https://doi.org/10.1371/journal.pone.0072923.

Claims (4)

1. Hexapeptide of general formula I X-Phe-Trp-Ala-His-Lys-Lys-Z (I) where: X is a -NHX ¹ phenylalanine group at the N-terminal end of the hexapeptide, wherein X ¹ is selected from the group consisting of H, acetyl, octanoyl, decanoyl, lauroyl, myristoyl, palmitoyl, stearoyl, elaidoyl, oleoyl, biotinoyl, or lipoyl; Z is a -COZ ¹ lysine group at the C-terminal end of the hexapeptide, where Z ¹ is selected from the group consisting of OH, OCH 3 , OCH 2 CH 3 or NH 2 . 2. Cosmetic topical composition, characterized in that it includes at least one hexapeptide according to any one of claims 1 to 3. 3. Pharmaceutical topical and/or cosmetic topical composition according to claim 9, characterized in that the auxiliary active ingredient is selected from the group including vitamins A, D, E, K, C, B-group of vitamins, coenzyme Q10, allantoin, bisabolol, bakuchiol, resveratrol, lactic acid, amino acids, benzoyl peroxide, sulfur, azelaic acid, peptides preferably acetyl hexapeptide-8, palmitoyl tripeptide-1, palmitoyl tetrapeptide-7, copper tripeptide-1, hexapeptide-1, palmitoyl pentapeptide-4, peptides saccharomyces, and proteins, preferably rice, soy, quinoa or wheat protein; polysaccharides, preferably hyaluronic acid or its pharmaceutically and cosmetically acceptable salt or derivative, preferably sodium oleoyl hyaluronate, sodium retinoyl hyaluronate, crosspolymer-3 sodium hyaluronate; carboxymethyl glucan, schizophyllan, glucomannan; panthenol; urea; glycerine; plant extracts, preferably aloe vera extract, chamomile extracts, acai extract, green tea extract, algae extract, oat extract, hemp extract, cranberry extract; bacteria preferably Lactobacillus spp., Thalassospira spp., Bifidobacterium spp., Halobacterium spp. or mushroom extracts from Saccharomyces, Agaricus subrufescens, Choiromyces meandriformis, Cordyceps sinensis, Ganoderma lucidum, Grifola frondosa, Hypsizygus ulmarium, Inonotus obliquus, Lentinula edodes, Polyporus spp., Trametes versicolor, Tremella fucformis, Tuber spp., Schizophyllum commune. 4. Pharmaceutical topical and/or cosmetic topical composition according to claim 14, characterized in that it further includes at least one cosmetic or dermo-pharmaceutical auxiliary component selected from the group comprising a thickener, an auxiliary active component and a preservative, as defined in claim 13.