Peptides and cosmetic compositions for use in the treatment of acne and its related skin conditions

Abstract

The present invention refers to peptides which provide antiacne and anti-inflammatory activities and, hence, are useful in cosmetics for the prevention, reduction and / or treatment of skin acne alterations as well as the overall improvement of skin quality. The invention also refers to the peptides for use as in medicine for the treatment of acne-related diseases.

Description

[0001] PEPTIDES AND COSMETIC COMPOSITIONS FOR USE IN THE TREATMENT OF ACNE AND ITS RELATED SKIN CONDITIONS

[0002] FIELD OF THE INVENTION

[0003] The present invention relates to the field of molecular biology, more precisely to molecular biology applied to cosmetics and medicine, even more precisely to peptides and compositions comprising said peptides able for the prevention and / or treatment of acne and its related skin conditions such as inflammation, redness, dryness, roughness and / or wrinkles.

[0004] BACKGROUND OF THE INVENTION

[0005] Skin conditions vary greatly in symptoms and severity. They can be temporary or permanent and may be painless or painful. Some have situational causes, while others may be genetic. Some skin conditions are minor, and others can be serious. Some well-known examples are sensitive skin, cellulite, acne (often combined to sebum overproduction, inflammation and enlarged pores), aging and stretch marks.

[0006] Acne, also known as Acne vulgaris, is a chronic inflammatory skin condition that occurs when dead skin cells and oil from the skin clog hair follicles. It primarily affects skin with a relatively high number of sebaceous glands, including the face, upper part of the chest, and back.

[0007] Acne vulgaris is a disorder with complex symptoms. Four factors play vital roles in acne pathophysiology:

[0008] Sebaceous gland hyperplasia (hyper-seborrhea and dys-seborrhea, sebum overproduction);

[0009] - Abnormal follicular differentiation with increased keratinization of the pilosebaceous duct;

[0010] Microbial hypercolonization of the follicular canal (most commonly Cutibacterium acnes or Propionibacterium acnes

[0011] Increased inflammation;

[0012] Extracellular Matrix (ECM) degradation. Excessive sebum production and accumulation and blocked sebaceous gland openings favor bacterial colonization. Due to the presence of lipase, these microorganisms hydrolyse sebum di- and triglycerides to free fatty acids. Free fatty acids, which arise during the hydrolysis process, have the irritating, proinflammatory effect and intensify follicular keratosis. Additionally, hyaluronidase, proteases and neuraminidases produced by Propionibacterium acnes (currently known as Cutibacterium acnes), have the pro- inflammatory effect. Moreover, the inflammatory condition of the tissue causes an overproduction of skin proteases leading to a further degradation of collagen and other ECM proteins in the dermis.

[0013] Inflammatory phenomena play a fundamental role in acne since they are responsible for the progression of the condition, in particular in the formation of inflammatory lesions as well as the occurrence of scars. This inflammatory process is multifactorial and the direct or indirect result of the proliferation of C. acnes. This bacterium is a commensal seed of the skin microflora. In normal skin, C. acnes develops at the bottom of the pilo-sebaceous follicle and wins the epidermal surface by sebum. During acne, the abnormal accumulation of corneocytes and the excess sebum in the follicle canal represents an ideal environment for the bacterial development. Its multiplication leads to the inflammatory process during the condition.

[0014] Conventional products for the treatment of oily and blemished skin, e.g., aqueous / ethanolic and / or surfactant-containing cleaning products have the disadvantage to strain the skin to dry out. It is particularly disadvantageous that the body's reaction to the use of ethanol and I or detergents-containing solutions over a longer period of time may be an overproduction of sebum and skin sensitivity, which counteracts the primary therapeutic goal of sebum reduction in oily, blemished and acne prone skin.

[0015] For the topical treatment of acne there are acne agents, such as strong oxidizing agents, such as benzoyl peroxide; alpha-hydroxy acids, such as salicylic acid and lactic acid; aliphatic dicarboxylic acids such as azelaic acid; retinoids such as tretinoin, all-trans-retinal and 13-cis-retinoic acid (isotretinoin); antiandrogens (5-alpha-reductase) and antibiotics such as clindamycin, tetracycline and erythromycin. These substances usually have only a very moderate antimicrobial activity against C. acnes and have to be used in a relatively high concentration, benzoyl peroxide for example with up to 5% wt and azelaic acid with up to 20% wt in cosmetic and dermatological formulations. Due to the high dosage, however, the skin is extremely disadvantageous heavily overused, which is manifested particularly in very dry skin, which is often associated in part with severe skin irritation due to the strong reduction of the pH of the skin.

[0016] Moreover, 13-c / s retinoic acid (ACCUTANE®) is recognized as teratogen and well known to produce significant systemic adverse events including risk of mental depression, increased blood lipid levels and deleterious mucocutaneous changes. Indeed, 13-c / s retinoic acid presents a number of serious adverse effects. The agent has a Pregnancy Category X teratogenicity warning and requires special prescribing precautions and routine pregnancy testing. Additionally, 13-c / s retinoic acid causes severe mucocutaneous toleration issues (dry skin, eyes, nasal passages, lips, etc.) which can be dose limiting if not adequately managed with palliative care. 13-c / s retinoic acid treatment is associated with adverse plasma lipid changes (increased TG, LDL) and hepatic toxicity (ALT / AST elevation requiring liver function testing prior to treatment. Additionally, 13-c / s retinoic acid therapy has also been associated with myalgia (50% of patients have elevated OK levels), calcification of ligaments and detrimental ocular effects (loss of night vision, loss of color vision and eye dryness). In isolated cases, 13-cis retinoic acid has been associated with neurological / psychological adverse effects including depression, psychosis and potentially suicide.

[0017] On other hand, hormonal therapies, including oral contraceptives and androgen receptor blockers, are used in female patients only for the treatment of moderate to severe acne with modest efficacy.

[0018] Bakuchiol is a natural ingredient from the leaves and seeds of the plant Psoralea corylifolia (or babchi). Bakuchiol has been disclosed as having soothing, anti-wrinkle, emollient and antimicrobial properties. And although it presents much less side-effects than retinol, there is still risk of sensitivity, including redness, dryness or stinging of the skin, especially with its initial use (Dhaliwal S et al, 2019. Prospective, randomized, double-blind assessment of topical bakuchiol and retinol for facial photoageing, British Journal of Dermatology, 180(2); 289-296).

[0019] Therefore, a need exists for a novel approach to treating acne with a favorable efficacy / safety profile. The problem is solved by a cosmetic or dermatological composition comprising specific peptides.

[0020] Peptides can be incorporated in cosmetic formulas. Bioactive peptides can imitate body’s own molecules and influence processes, with the advantage that they have much better tolerability and stability. In addition, a wide range of activities, chemistries and indications can be developed for them (Zhang L and Falla TJ, 2009. Cosmeceuticals and peptides. Clinics in dermatology, 27, 485-494).

[0021] The insulin-like growth factor 1 receptor (IGF1 R) is a protein found on the surface of human cells. It is a transmembrane receptor activated by the insulin-like growth factor 1 (IGF1) and by IGF2. Evidence underlines the role of an increased IGF1 signaling in the pathogenesis of acne. The peak incidence of acne occurs during adolescence, when serum IGF1 levels reach their peak, and IGF1 levels showed a positive correlation with the severity of acne in women with clinical acne. Moreover, increased serum levels of IGF1 and number of total acne lesions have been observed in men with acne (Makrantonaki E et al., 2011. An update on the role of the sebaceous gland in the pathogenesis of acne. Dermatoendocrinol 3(1); 41-49) as well as in menopausal females due to hormonal imbalance (Khunger N and Mehrotra K, 2019. Menopausal acne - Challenges and solutions. Int J Womens Health 11 ; 555-567). IGF1 and IGF1 R overexpression in both acne lesions and skin explants was associated with an increase in Ki67 and filaggrin expression in the epidermis, confirming that the IGF1 / IGF1 R system is associated with the modulation of both proliferation and differentiation of keratinocytes. It was previously noted that Ki67 expression is higher in acne patients’ epidermis than in normal healthy skin (Knaggs et al., 1994. Quantification of cellular proliferation in acne using the monoclonal antibody Ki-67. J Invest Dermatol, 102, pp. 89-92).

[0022] Acetyl-CoA carboxylase (ACC) is known to be highly expressed in human sebaceous glands and catalyzes the first rate-limiting step in the biosynthesis of fatty acids (i.e., acetyl- CoA to malonyl-CoA) that corresponds to the 80% of human sebum. Excess sebum production contributes to the development of acne, and suppression of sebum production reduces acne incidence and severity. Therefore, inhibition of ACC is a rational approach for affecting sebaceous gland activity and thus inhibiting an important factor of acne.

[0023] Matrix metalloproteases are produced by different types of cells, including keratinocytes, fibroblasts and sebocytes. They play important roles in acne inflammation, dermal matrix degradation and hyperproliferative skin disorders. MMPs also cause rupture of the pilosebaceous follicle to exacerbate inflammation. MMPs are involved in normal tissue repair and are highly upregulated in many inflammatory states, including acne. Studies have shown that C. acnes triggers the increased production and activity of several MMPs. A few types of MMPs were found in the sebum of acne patients, including MMP1 , MMP13, and MMP9. Follicular hyperkeratinization is a key element in the pathogenesis of acne and a main target of retinoid activity. Hyperkeratinization occurs when the cells of the follicle become cohesive and do not shed normally onto the skin’s surface. The result is a microcomedone (the precursor to acne) and subsequent lesions characteristic of acne. IGF1 and IGF1 R overexpression in both acne lesions and skin explants was associated with an increase in Ki67 and filaggrin expression in the epidermis, confirming that the IGF1 / IGF1 R system is associated with the modulation of both proliferation and differentiation of keratinocytes. Ki67 is a nuclear protein associated with cellular proliferation and necessary for the maintenance of cell proliferation (Heenen et al., 1998. Ki-67 immunostaining of normal human epidermis: comparison with 3H-thymidine labelling and PCNA immunostaining. Dermatology 197(2): 123-6). It was previously noted that Ki67 expression is higher in acne patients’ epidermis than in normal healthy skin (Knaggs et al., 1994). GATA6, which is expressed in the upper pilosebaceous unit of normal human skin, is downregulated in acne. GATA6 controls keratinocyte proliferation and differentiation to prevent hyperkeratinization of the infundibulum, which is the primary pathological event in acne. In an embodiment, the peptides of the invention are able to decrease Ki67 and increase GATA6.

[0024] Liver X receptor (LXR) was identified as an orphan nuclear receptor that regulates the metabolism of lipid and cholesterol. Activation of LXRa induces lipid synthesis in sebocytes. Additionally, it induces Sterol Regulatory Element-Binding Protein 1 (SREBP1) that regulates genes required for fatty acid and lipid metabolism and production. IGF1 stimulates the lipogenic transcription factor sterol regulatory element-binding protein-1 (SREBP1) which in turn plays a fundamental role in lipogenesis. In addition, SREBP1 regulates Acetyl- CoA Carboxylase (ACC) gene.

[0025] The inventors of the present invention, after extensive and exhaustive research, have surprisingly found peptides which reduce the activity of IGF1 R, and / or ACC and / or MMP1 . These peptides have anti-acne, anti-sebum, anti-inflammatory and anti-aging activities. The peptides of the present invention are, therefore, useful for the treatment of skin acne and related conditions, hence, solve the problem present in the state of the art mentioned above.

[0026] DESCRIPTION OF THE INVENTION

[0027] BRIEF DESCRIPTION OF THE INVENTION

[0028] The invention is defined in the appended claims. In a first aspect, the present invention refers to a peptide of formula (I):

[0029] Ri-AA1-AA2-AA3-AA4-AA5-AA6 -R2

[0030] (I) its acceptable isomers, salts, solvates and / or mixtures thereof, wherein:

[0031] AA1 is absent or is an amino acid selected from Trp, Arg, His, Gin, Lys, Ser and Tyr;

[0032] AA2 is an amino acid selected from Lys, His, Trp, Glu, His and Pro;

[0033] AA3 is an amino acid selected from Ser, Leu Tyr, Glu, Gin, Arg, Trp, Ala, Thr, and Leu;

[0034] AA4 is an amino acid selected from Gin, Tyr, Asp, Glu, Pro, Lys, Leu, His, and Arg;

[0035] AA5 is selected from Trp, Gin, Vai, Thr, Lys, Arg, Leu, His, Phe, and Ala; and

[0036] AA6 is absent or is an amino acid selected from His, Ser, Glu, Tyr, Lys, Phe, Leu or Ala.

[0037] Ri is selected from H, substituted or unsubstituted non-cyclic aliphatic, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl and Rs-CO-, wherein R5 is selected from the group formed by substituted or unsubstituted C1-C24 alkyl radical, substituted or unsubstituted C2-C24 alkenyl, substituted or unsubstituted C2-C24 alkynyl, substituted or unsubstituted C3-C24 cycloalkyl, substituted or unsubstituted C5-C24 cycloalkenyl, substituted or unsubstituted C8-C24 cycloalkynyl, substituted or unsubstituted C6-C30 aryl, substituted or unsubstituted C7-C24 aralkyl, substituted or unsubstituted heterocyclyl ring of 3 to 10 members, and substituted or unsubstituted heteroarylalkyl of 2 to 24 carbon atoms and 1 to 3 atoms other than carbon and an alkyl chain of 1 to 6 carbon atoms; and

[0038] R2 is selected from H, -NR3R4- , -OR3 and -SR3, wherein R3 and R4 are independently selected from H, substituted or unsubstituted non-cyclic aliphatic group, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

[0039] The second aspect of the invention is directed to a cosmetic composition comprising the peptide of the invention.

[0040] The third aspect of the invention is directed to a pharmaceutical composition comprising the peptide of the invention. The fourth aspect of the invention is directed to the use of the peptide as an ingredient in a cosmetic composition.

[0041] The fifth aspect of the invention is directed to the peptide, or the pharmaceutical composition disclosed above, for use as a medicament.

[0042] The sixth aspect of the invention is directed to the peptide, or the pharmaceutical composition disclosed above, for use in the prevention and / or treatment of acne in a subject.

[0043] The seventh aspect of the invention is directed to the peptide, or the pharmaceutical composition disclosed above, for use in the prevention and / or treatment of an acne-related disease and / or a skin inflammatory disease.

[0044] BRIEF DESCRIPTION OF THE FIGURES

[0045] Figure 1. Bar Graph represents the percentage of activity of ACC after treatment with different concentrations of SEQ ID NO: 21 (A), SEQ ID NO: 20 (B), SEQ ID NO: 6 (C) and SEQ ID NO: 3 (D). Graphic shows the mean ± standard error of the mean (SEM) (N=3 and n=3).

[0046] Figure 2. Bar Graph represents the percentage of activity of MMP1 after treatment with different concentrations of SEQ ID NO: 21 (A), SEQ ID NO: 3 (B), SEQ ID NO: 19 (C) and SEQ ID NO: 20 (D). Graphic shows the mean ± standard error of the mean (SEM) (N=3 and n=3).

[0047] Figure 3. Bar Graph represents the percentage of phosphorylation of IGF1 R after treatment with SEQ ID NO: 21 at 0.005, 00.1 , 0.05, 0.1 or 0.5 mg / mL at six hours, normalized with BCA.

[0048] Figure 4. Quantification of the images acquired with a fluorescence microscope in the BODIPY assay on Human Sebocytes after 48 hours of treatment with SEQ ID NO: 21. Graphic shows the mean ± standard error of the mean (SEM).

[0049] Figure 5. Pan-keratin stained area on sebocytes after 48 hours of treatment with SEQ ID NO: 21. Fluorescence quantification of the images acquired with a fluorescence microscope. Graphic shows the mean ± standard error of the mean (SEM). Figure 6. Modulation of several sebum-related genes in sebocytes treated with SEQ ID NO: 21 at 0.01 mg / mL for 24 h (IGF1 R, ACACA, ACACB and TLR2) or 0.005 mg / mL during 6 h (NR1 H3 and PTGS2) or 24 h (FOXO1 and MUC1).

[0050] Figure 7. NR1 H3 immunostaining on sebocytes after 24 hours of treatment with SEQ ID NO: 21 at 0.005 mg / mL. Fluorescence quantification of the images acquired with a fluorescence microscope. Graphic shows the mean ± standard error of the mean (SEM).

[0051] Figure 8. Modulation of several acne-related genes in HEKa treated with SEQ ID NO: 21 at 0.005 mg / mL for 24 h.

[0052] Figure 9. Modulation of several acne-related genes in HEKa treated with 107CFUs of C. acnes alone or in the presence of SEQ ID NO: 21 at 0.005 mg / mL for 24 h. Fold changes were compared vs. cells untreated (Basal). Light grey: C. acnes, dark grey: C. acnes + SEQ ID NO: 21.

[0053] Figure 10. IL1a release on HEKa cells after treatment with C. acnes alone or in the presence of 0.01 or 0.05 mg / mL of SEQ ID NO: 21 (normalized). Fluorescence quantification of the images acquired with a fluorescence microscope. Graphic shows the mean ± standard error of the mean (SEM).

[0054] Figure 11. Ki67 positive cells after 6 hours of treatment with C. acnes alone or in the presence of 0.01 or 0.05 mg / mL of SEQ ID NO: 21 (normalized). Fluorescence quantification of the images acquired with a fluorescence microscope. Graphic shows the mean ± standard error of the mean (SEM).

[0055] Figure 12. (A) Histogram representation of the elastic modulus of the matrix surrounding the pores after 3 days of treatment. The matrix surrounding 9 skin pores was analyzed. Statistical t-test. ****: p-value < 0.00005. R-studio software. Left bar: NT, right bar: Active. (B). Histogram representation of the diameter of the skin pores at depths 150 pm and 500 pm after 3 days of treatment and table with data. Statistical Wilcoxon and T-tests. ns: nonsignificant, *: p-value<0.05. R-studio software.

[0056] Figure 13. Skin serum content. (A) Mean data obtained at each experimental time for the analyzed skin parameter. The data are expressed as a mean ± SE. (B) Mean % variations obtained at each timepoint for the analyzed skin parameter. Black bars correspond to the sample without the active ingredient (control), whereas white bars are the control with the peptide. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO). The above asterisks in (A): * p<0.05, ** p<0.01 , *** p<0.001 inter-group statistical analysis (Product A vs Product B, carried out on % variations).

[0057] Figure 14. Mean data obtained at each experimental time for the analyzed skin parameter (A). The data are expressed as a mean ± SE. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO). Upper asterisks: * p<0.05, ** p<0.01 , *** p<0.001 inter-group statistical analysis (Product A vs Product B, carried out on % variations). The graph (B) reports the mean % variations obtained at each timepoint for the analyzed skin parameter. Black bars correspond to the product without peptide, and white bars correspond to the product with the peptide.

[0058] Figure 15. (A) Number of inflammatory lesions at 0, 7, 14 and 28 days treated by the composition comprising the peptide (white bars) and the control without peptide (black bars). (B) Number of total lesions. (C) % variation of the inflammatory lesions at each time point in relation to TO. (D) % variation of the total number of lesions at each time point in relation to TO. All data are expressed as mean data obtained at each experimental time for the analyzed skin parameter. The data are expressed as a mean ± SE. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO).

[0059] Figure 16. (A) Mean data obtained at each experimental time for the total area occupied by protoporphyrin after treatment with the composition comprising the peptide (white bars) and the control without peptide (black bars). The data are expressed as a mean ± SE. Total area occupied by protoporphyrin at TO, T7; T14 and T24. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO) and the above Asterisk in T28 shows inter- group statistical analysis (Product A vs Product B, carried out on % variations). (B) mean % variations obtained at each timepoint for the analyzed parameter.

[0060] Figure 17. (A) Mean data obtained at each experimental time for the skin moisturization after treatment with the composition comprising the peptide (white bars) and the control without peptide (black bars). The data are expressed as a mean ± SE. Moisturization at TO, T7; T14 and T24. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO). (B) mean % variations obtained at each timepoint for the analyzed parameter.

[0061] Figure 18. (A) Mean data obtained at each experimental time for the LPO (MDA pM) after treatment with the composition comprising the peptide (white bars) and the control without peptide (black bars). The data are expressed as a mean ± SE. Total area occupied by protoporphyrin at TO and T24. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO) and the above Asterisk in T28 shows inter-group statistical analysis (Product A vs Product B, carried out on % variations). (B) mean % variations obtained at each timepoint for the analyzed parameter.

[0062] DETAILED DESCRIPTION OF THE INVENTION

[0063] The present disclosure describes peptides and compositions comprising thereof useful in the treatment of acne and its related skin conditions, such as inflammation, redness, dryness, roughness and / or wrinkles.

[0064] The present disclosure is directed toward peptides able to inhibit dimerization of ACC and / or inhibition of IGF1 R activation and / or inhibition of MMP1 activity. Preferably, the peptides are able to inhibit dimerization of ACC, inhibit IGF1 R activation and inhibit MMP1 activity. It has been shown that these activities reduce lipid synthesis, decrease inflammation and improve ECM structural integrity, which lead to a reduction of acne, sebum overproduction and / or enlarged pores in a subject.

[0065] The data disclosed herein proves that the claimed peptides are able to:

[0066] Inhibit IGF1 R activation, which reduces lipid synthesis and blocks inflammation;

[0067] Inhibit ACC dimerization, which reduces fatty acid biosynthesis; and Inhibit MMP1 , which improves ECM structural integrity.

[0068] These effects result in pore reduction, sebum control, reduction of acne and general improvement of skin quality. Skin quality includes hydration, luminosity and less roughness and wrinkles.

[0069] In a first aspect, the present invention refers to a peptide of formula (I): Ri-AA1-AA2-AA3-AA4-AA5-AA6 -R2

[0070] (I) its acceptable isomers, salts, solvates and / or mixtures thereof, wherein:

[0071] AA1 is absent or is an amino acid selected from Trp, Arg, His, Gin, Lys, Ser and Tyr;

[0072] AA2 is an amino acid selected from Lys, His, Trp, Glu, His and Pro;

[0073] AA3 is an amino acid selected from Ser, Leu Tyr, Glu, Gin, Arg, Trp, Ala, Thr, and Leu;

[0074] AA4 is an amino acid selected from Gin, Tyr, Asp, Glu, Pro, Lys, Leu, His, and Arg;

[0075] AA5 is selected from Trp, Gin, Vai, Thr, Lys, Arg, Leu, His, Phe, and Ala; and AA6 is absent or is an amino acid selected from His, Ser, Glu, Tyr, Lys, Phe, Leu or Ala.

[0076] Ri is selected from H, substituted or unsubstituted non-cyclic aliphatic, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl and Rs-CO-, wherein R5 is selected from the group formed by substituted or unsubstituted C1-C24 alkyl radical, substituted or unsubstituted C2-C24 alkenyl, substituted or unsubstituted C2-C24 alkynyl, substituted or unsubstituted C3-C24 cycloalkyl, substituted or unsubstituted C5-C24 cycloalkenyl, substituted or unsubstituted C8-C24 cycloalkynyl, substituted or unsubstituted C6-C30 aryl, substituted or unsubstituted C7-C24 aralkyl, substituted or unsubstituted heterocyclyl ring of 3 to 10 members, and substituted or unsubstituted heteroarylalkyl of 2 to 24 carbon atoms and 1 to 3 atoms other than carbon and an alkyl chain of 1 to 6 carbon atoms; and

[0077] R2 is selected from H, -NR3R4- , -OR3 and -SR3, wherein R3 and R4 are independently selected from H, substituted or unsubstituted non-cyclic aliphatic group, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl; alternatively, R1 and R2 are absent and AA1 is directly bound to the last amino acid of the peptide forming a cyclic peptide.

[0078] R1 is linked to the N-terminal of the peptide, this is, to the amino group of AA1 (it substitutes or replaces an H from the amino group of AA1).

[0079] R2 is linked to the C-terminal of the peptide, this is, to the carboxy group of AA6 (it substitutes or replaces the OH of the carboxy group).

[0080] It is contemplated that the amino acids used or present in the peptides of the present invention are L-amino acids, D-amino acids or combinations thereof. In a preferred embodiment, the amino acids used or present in the peptides of the present invention are L-amino acids.

[0081] Preferably, the isomers mentioned above are stereoisomers. It is contemplated that said stereoisomers are enantiomers or diastereoisomers. Hence, in a preferred embodiment of the present invention, the peptide is a racemic mixture, a diastereomeric mixture, a pure enantiomer or a pure diastereoisomer.

[0082] In addition, isomers, salts, solvates and / or mixtures thereof are, preferably, cosmetically, pharmaceutically and / or food acceptable isomers, salts, solvates, derivatives and / or mixtures thereof.

[0083] In a preferred embodiment, the peptide interacts with ACC, MMP1 and / or IGF1 R.

[0084] In a more preferred embodiment, the peptide of the present invention is selected from: Ri-Lys-Tyr-Asp-Val-R2(SEQ ID NO: 3);

[0085] Ri-Trp-His-Tyr-Glu-Arg-R2(SEQ ID NO: 6);

[0086] Ri-Arg-Trp-Ala-His-Phe-Leu-R2(SEQ ID NO: 19);

[0087] Ri-Arg-Trp-leu-His-Phe-Ala-R2(SEQ ID NO: 20); and Ri-His-Trp-Leu-Arg-Ala-Phe-R2(SEQ ID NO: 21), being Ri and R2as disclosed above.

[0088] More preferably, the peptide of the present invention is selected from:

[0089] Ri-Lys-Tyr-Asp-Val-R2(SEQ ID NO: 3)

[0090] Ri-Arg-Trp-Ala-His-Phe-Leu-R2(SEQ ID NO: 19)

[0091] Ri-Arg-Trp-Leu-His-Phe-Ala-R2(SEQ ID NO: 20) or

[0092] Ri-His-Trp-Leu-Arg-Ala-Phe-R2(SEQ ID NO: 21).

[0093] The above peptides interact with the three targets selected, responsible of the acne pathology (ACC, MMP1 and IGF1 R).

[0094] In any of the above embodiments, preferably, Ri is H or acetyl, more preferably Ri is acetyl. Also, in any of the above embodiments, R2is NH2or OH, more preferably R2is NH2. Hence, in a more preferred embodiment, Ri is acetyl and R2is NH2.

[0095] In the more preferred embodiment, the peptide of the present invention is:

[0096] Ac-Lys-Tyr-Asp-Val-NH2(SEQ ID NO: 3);

[0097] Ac-Arg-Trp-Ala-His-Phe-Leu-NH2(SEQ ID NO: 19);

[0098] Ac-Arg-Trp-leu-His-Phe-Ala-NH2(SEQ ID NO: 20); and Ac-His-Trp-Leu-Arg-Ala-Phe-NH2(SEQ ID NO: 21).

[0099] The term "non-cyclic aliphatic group" and its plural, as used herein, have the common meaning given in the state of the art to said terms. Therefore, these terms refer to, for example and not restricted to, linear or branched alkyl, alkenyl and alkynyl groups.

[0100] The term "alkyl group" and its plural, as used herein, refer to a saturated, linear or branched group, which has between 1 and 24, preferably between 1 and 16, more preferably between 1 and 14, even more preferably between 1 and 12, and even more preferably still between 1 , 2, 3, 4, 5 or 6 carbon atoms and which is bound to the rest of the molecule by a simple bond, including, for example and not restricted to, methyl, ethyl, isopropyl, n-propyl, i-propyl, isobutyl, tert-butyl, n-butyl, sec-butyl, n-pentyl, n-hexyl, heptyl, octyl, decyl, dodecyl, lauryl, hexadecyl, octadecyl, amyl, 2-ethylhexyl, 2-methylbutyl, 5-methylhexyl and similar. The alkyl groups can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0101] The term "alkenyl group" and its plural, as used herein, refer to a linear or branched group which has between 2 and 24, preferably between 2 and 16, more preferably between 2 and 14, even more preferably between 2 and 12, even more preferably still 2, 3, 4, 5 or 6 carbon atoms, with one or more carbon-carbon double bonds, preferably with 1 , 2 or 3 carboncarbon double bonds, conjugated or unconjugated, which is bound to the rest of the molecule through a single bond, including, for example and not restricted to, the vinyl, oleyl, linoleyl and similar groups. The alkenyl groups can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxycarbonyl, amino, nitro, mercapto and alkoxythio.

[0102] The term "alkynyl group" and its plural, as used herein, refer to a linear or branched group which has between 2 and 24, preferably between 2 and 16, more preferably between 2 and 14, even more preferably between 2 and 12, even more preferably still 2, 3, 4, 5 or 6 carbon atoms, with one or more carbon-carbon triple bonds, preferably with 1 , 2 or 3 carbon-carbon triple bonds, conjugated or unconjugated, which is bound to the rest of the molecule through a single bond, including, for example and not restricted to, the ethinyl group, 1 -propinyl, 2- propinyl, 1 -butinyl, 2-butinyl, 3-butinyl, pentinyl, such as 1 -pentinyl and similar groups. The alkynyl groups can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0103] The term "alicyclic group" and its plural, as used herein, have the common meaning given in the state of the art to said terms. Hence, these terms are used to refer to, for example and not restricted to, cycloalkyl or cycloalkenyl or cycloalkynyl groups.

[0104] The term "cycloalkyl" and its plural, as used herein, refer to a saturated mono- or polycyclic aliphatic group which has between 3 and 24, preferably between 3 and 16, more preferably between 3 and 14, even more preferably between 3 and 12, even more preferably still 3, 4, 5 or 6 carbon atoms and which is bound to the rest of the molecule through a single bond, including, for example and not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, methyl cyclohexyl, dimethyl cyclohexyl, octahydroindene, decahydronaphthalene, dodecahydro-phenalene, adamantyl and similar, and that can optionally be substituted by one or more groups, such as, alkyl, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0105] The term "cycloalkenyl" and its plural, as used herein, refer to a non-aromatic mono- or polycyclic aliphatic group which has between 5 and 24, preferably between 5 and 16, more preferably between 5 and 14, even more preferably between 5 and 12, even more preferably still 5 or 6 carbon atoms, with one or more carbon-carbon double bonds, preferably with 1 , 2 or 3 carbon-carbon double bonds, conjugated or unconjugated, which is bound to the rest of the molecule through a single bond, including, for example and not restricted to, the cyclopent-1 -en-1-yl group and similar groups, and that can optionally be substituted by one or more groups, such as, alkyl, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0106] The term "cycloalkynyl" and its plural, as used herein, refer to a non-aromatic mono- or polycyclic aliphatic group which has between 8 and 24, preferably between 8 and 16, more preferably between 8 and 14, even more preferably between 8 and 12, even more preferably still 8 or 9 carbon atoms, with one or more carbon-carbon triple bonds, preferably with 1 , 2 or 3 carbon-carbon triple bonds, conjugated or unconjugated, which is bound to the rest of the molecule through a single bond, including, for example and not restricted to, the cyclooct-2-yn-1-yl group and similar, and that can optionally be substituted by one or more groups, such as, alkyl, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxycarbonyl, amino, nitro, mercapto and alkoxythio.

[0107] The term "aryl group" and its plural, as used herein, refer to an aromatic group which has between 6 and 30, preferably between 6 and 18, more preferably between 6 and 10, even more preferably 6 or 10 carbon atoms, which comprises 1 , 2, 3 or 4 aromatic rings, bound by a carbon-carbon bond or fused, and which is bound to the rest of the molecule through a single bond, including, for example and not restricted to, phenyl, naphthyl, diphenyl, indenyl, phenanthryl or anthranyl among others. The aryl group can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0108] The term "aralkyl group" and its plural, as used herein, refer to an alkyl group substituted by an aromatic group, with between 7 and 24 carbon atoms and including, for example and not restricted to, -(CH2)1-6-phenyl, -(CH2)1-6-(1-naphtyl), -(CH2)1-6-(2-naphtyl), -(CH2)1-6- CH(phenyl)2 and similar. The aralkyl groups can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxycarbonyl, amino, nitro, mercapto and alkoxythio.

[0109] The term "heterocyclic group" and its plural, as used herein, refer to a 3-10-member heterocycyl or hydrocarbon ring, in which one or more of the ring atoms, preferably 1 , 2 or 3 of the ring atoms, is a different element to carbon, such as nitrogen, oxygen or sulfur and may be saturated or unsaturated. For the purposes of this invention, the heterocyclyl can be a cyclic, monocyclic, bicyclic or tricyclic system which may include fused ring systems; and the nitrogen, carbon or sulfur atoms can be optionally oxidized in the heterocyclyl radical; the nitrogen atom can optionally be quaternized; and the heterocyclyl radical may be partially or completely saturated or may be aromatic. With increasing preference, the term heterocyclic relates to a 5 or6-member ring. The heterocyclic groups can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0110] The term "heteroarylalkyl group" and its plural, as used herein, refer to an alkyl group substituted with a substituted or unsubstituted aromatic heterocyclyl group, the alkyl group having from 1 to 6 carbon atoms and the aromatic heterocyclyl group between 2 and 24 carbon atoms and from 1 to 3 atoms other than carbon and including, for example and not restricted to, -(CH2)1-6-imidazolyl, -(CH2)1-6-triazolyl, -(CH2)1-6-thienyl, -(CH2)1-6-furyl, - (CH2)1-6-pyrrolidinyl and similar. The heteroarylalkyl groups can be optionally substituted by one or more substituents, such as, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0111] The terms "halo" or "halogen", as used in the present document, refer to fluorine, chlorine, bromine or iodine, and its anions are referred to as halides.

[0112] As used in the present document, the term "salt" and its plurals refer to any type of salt from among those known in the state of the art, for example, halide salts, hydroxy acid salts (such as oxyacid salts, acid salts, basic salts and double salts), hydroxo salts, mixed salts, oxy salts or other hydrated salts. This term comprises cosmetically, pharmaceutically and / or food acceptable salts; and cosmetically, pharmaceutically and / or food unacceptable salts, since the latter may be useful in the preparation of cosmetically, pharmaceutically and / or food acceptable salts.

[0113] As used in the present document, the term "isomer" and its plural refer to optical isomers, enantiomers, stereoisomers or diastereoisomers. The individual enantiomers or diastereoisomers, as well as their mixtures, may be separated by conventional techniques known in the state of the art.

[0114] As used herein, the term "solvate" and its plural refer to any solvate known in the state of the art, such as polar, apolar or amphiphilic solvates, and include any cosmetically, pharmaceutically and / or food acceptable solvate which, when administered or applied to the interested subject (directly or indirectly) provides the compound of interest (the peptide or peptides of the present invention). Preferably, the solvate is a hydrate, a solvate with an alcohol such as methanol, ethanol, propanol or isopropanol, a solvate with an ester such as ethyl acetate, a solvate with an ether such as methyl ether, ethyl ether or THF (tetrahydrofuran) or a solvate with DMF (dimethylformamide), and more preferably a hydrate or a solvate with an alcohol such as ethanol.

[0115] In addition, as used herein, the term “amino acid” and its plural include the amino acids codified by the genetic code as well as uncodified amino acids, whether they are natural or not and whether they are D- and L-amino acids. Examples of uncodified amino acids are, without restriction, citrulline, ornithine, sarcosine, desmosine, norvaline, 4-aminobutyric acid, 2-aminobutyric acid, 2-aminoisobutyric acid, 6-aminohexanoic acid, 1- naphthylalanine, 2-naphthylalanine, 2-aminobenzoic acid, 4 aminobenzoic acid, 4- chlorophenylalanine, 2,3-diaminopropionic acid, 2,4 diaminobutyric acid, cycloserine, carnitine, cysteine, penicillamine, pyroglutamic acid, thienylalanine, hydroxy proline, alloisoleucine, allo-threonine, isonipecotic acid, isoserine, phenylglycine, statin, p-alanine, norleucine, N-methylamino acids, a-amino acids and p-amino acids, among others, as well as their derivatives. Nevertheless, further unnatural amino acids are known in the state of the art (see, for example, “Unusual amino acids in peptide synthesis" by D. C. Roberts and F. Vellaccio, The Peptides, Vol. 5 (1983), Chapter VI, Gross E. and Meienhofer J., Eds., Academic Press, New York, USA). The abbreviations used in the present text for amino acids follow the 1983 IUPAC-IUB Joint Commission on Biochemical Nomenclature recommendations outlined in Eur. J. Biochem. (1984) 138:937. In a preferred embodiment, combinable with any of the above embodiments, the peptide of the present invention inhibits IGF1 R activation, ACC dimerization and MMP1 activity. These activities lead to pore reduction, sebum control, reduction of inflammation and overall improvement of skin quality, being therefore suitable for the treatment of acne and related skin conditions.

[0116] In a further preferred embodiment, combinable with any of the above embodiments, the peptide of the present invention provides for anti-inflammatory activity, preferably by means of modulation of inflammation-related genes, decreasing the levels of IL1 A, and / or reducing the levels of CXCL8 and / or reducing the levels of IL1 B or combinations thereof, most preferably by decreasing the levels of IL1 a protein.

[0117] Modulation of acne is generally due to the upregulation of genes such as FOXO1 (related to proliferation, inflammation and lipid synthesis) and GATA6 (related to keratinization), and downregulation of MMP1 , MMP9, ACC, IGF1 R and moreover MUC1 / EMA (related to sebocyte differentiation), PTGS2 / COX2, NR1 H3, (related to proliferation, inflammation and lipid synthesis), TLR2, IL1A, IL1 B, CXCL8, TNF (related to response to C. acnes and inflammation), IL1 A, and, KRT16 (related to abnormal keratinization).

[0118] Within the scope of the present invention are also included peptides derived or obtained from any of the peptides disclosed above, preferably incorporating one or more conservative substitutions, as long as they retain or show or improve at least one of the activities mentioned above (and, preferably, all of the activities mentioned above for the peptides of the present invention).

[0119] As it is directly derivable from the above, it is contemplated that the peptide of the present invention is a linear peptide or a cyclic peptide. Cyclization is known to be an effective strategy to enhance stability against enzymatic and chemical degradation. Cyclized (or cycled) peptides tend to have better permeability and stability because they are less prone to hydrolysis of exo- and endopeptidases. The elimination on the charged termini may enhances membrane permeability. These peptides may be used to synthesize analogues that mimic the biological activity of natural structures enhancing its bioavailability, reducing conformational flexibility that often lead to a better interaction with its target. In summary, cycled peptides may produce an improved biological activity over their linear forms. However, there is not information available of cycled peptides in compositions with an improved activity over their linear counterparts. The cyclization of peptides may be produced by:

[0120] - Head-to-tail (end-to-end, C-terminus to N-terminus) cyclization (amide bond formation between amino and carboxyl termini, many biologically active cyclic peptides are formed this way).

[0121] - End-to-side chain cyclization (requires a trifunctional amino acid, either head-to- side chain or side chain-to-tail).

[0122] - Side chain-to-side chain cyclization (requires 2 trifunctional amino acids).

[0123] (White C and Yudin A (2011). Contemporary strategies for peptide macrocyclization. Nature Chem 3, 509-524).

[0124] In an embodiment, which can be combined with any of the above embodiments, the peptide is cyclic. In this case, the first and the last amino acid of the peptide of the present invention (in accordance with the above explanations) are bound. Therefore, appropriate modification to any of the above embodiments has to be done (for example, Ri and R2 may be absent as the amino acids to which said moieties are bound, are now bound to each other).

[0125] The peptides of the present invention may be synthesized and produced by any means known in the state of the art. For example, they may be synthesized and produced by chemical synthesis (preferably, by means of solid phase peptide synthesis), expressing said peptides in cell cultures or by means of transgenic production of the peptide in plants or animals. In addition, the peptides of the present invention may be purified by any means known in the state of the art.

[0126] In a preferred embodiment, which can be combined with any of the embodiments disclosed above, the peptide of the present invention is suited or adapted to be applied by means of iontophoresis, more preferably, in the face of a subject (preferably, a human).

[0127] In another preferred embodiment, which can be combined with any of the embodiments disclosed above, the peptide of the present invention is suited or adapted to be applied topically (preferably, in the form of a cream), more preferably, in the face of a subject (preferably, a human).

[0128] As it can be directly derived from the examples included below, the peptides of the present invention provide for a broad spectrum of activities which make them useful in cosmetics for the prevention and / or treatment of acne and / or related inflammatory processes and / or other related skin conditions.

[0129] Therefore, the peptides of the present invention provide for a broad-spectrum activity for the prevention and / or treatment of acne and solve the problems present in the state of the art and mentioned above.

[0130] In addition, the peptides of the present invention may be also useful in medicine due to the above-mentioned activities (especially anti-inflammatory activity).

[0131] In a second aspect, the present invention refers to a composition comprising one or more peptides of the present invention.

[0132] The peptide or peptides of the composition are as disclosed above in the first aspect of the present invention.

[0133] The composition of the present invention is preferably a cosmetic composition. Hence, in this embodiment, the cosmetic composition of the present invention comprises a cosmetically effective amount of the at least one peptide of the present invention, more preferably the cosmetic composition of the present invention comprises from 0.0001 % (weight / volume in g / 100 mL, hereinafter, w / v) to 0.05% (w / v) of a peptide of the present invention. In a most preferred embodiment, the cosmetic composition of the present invention comprises from 0.0001% (w / v) to 0.005% (w / v) of a peptide of the present invention, more preferably, from 0.0005% (w / v) y 0.0015% (w / v).

[0134] It is contemplated that the cosmetic composition of the present invention also comprises at least one additional cosmetic ingredient. Said additional cosmetic ingredient can be at least one excipient and / or at least one additional cosmetic active ingredient. Said additional cosmetic ingredient can be any cosmetic ingredient known in the state of the art as long as it does not affect, or it does not affect in an unacceptable manner the activity of the peptides of the present invention.

[0135] In another embodiment, the composition of the present invention is a pharmaceutical composition.

[0136] It is contemplated that the pharmaceutical composition of the present invention also comprises at least one additional pharmaceutical ingredient. Said additional pharmaceutical ingredient can be at least one excipient and / or at least one additional pharmaceutical active ingredient. Said additional pharmaceutical ingredient can be any cosmetic ingredient known in the state of the art as long as it does not affect, or it does not affect in an unacceptable manner the activity of the peptides of the present invention.

[0137] In another embodiment, the composition of the present invention consists of peptides of the present invention.

[0138] In a third aspect, the present invention refers to the use as a cosmetic of one or more peptides of the first embodiment, or the use as a cosmetic of a cosmetic composition as disclosed above, in a subject in need of acne cosmetic treatment.

[0139] The peptide or peptides of this embodiment are disclosed in the first aspect of the invention.

[0140] The peptide or the composition of the present invention are used in a cosmetically effective amount or quantity.

[0141] The composition is, preferably, a cosmetic composition as disclosed above in the second aspect of the present invention.

[0142] In a preferred embodiment, the use as a cosmetic is to prevent, treat and / or reduce skin acne conditions and / or related skin conditions, such as inflammation, redness, dryness, roughness and / or wrinkles.

[0143] In this third aspect of the present invention, the subject in need of the treatment is preferably a mammal, more preferably, a human.

[0144] In a fourth aspect, as noted above, the present invention refers to the cosmetic use of a peptide of the present invention or a composition of the present invention in a subject in need of the treatment.

[0145] Preferably, in this fourth aspect of the present invention, the cosmetic use is to prevent and / or reduce signs of acne, such as, enlarged pores or sebum overproduction. In a more preferred embodiment, the use of the peptide or peptides is for one or more of pore reduction, sebum control, reduction of inflammation, redness, dryness, roughness, wrinkles and / or increase of hydration of the skin. In a fifth aspect, the present invention refers to a method for the prevention and / or treatment of acne in a subject comprising the administration of a peptide of the present invention or a composition of the present invention to the subject. Preferably, the administration is topical and / or iontophoretic.

[0146] In a sixth aspect, the present invention refers to a composition or at least one peptide of the present invention for use as a medicament. In this sixth aspect, the composition or at least one peptide, is administered to a subject in need of the treatment. Preferably, the subject in need of the treatment is a mammal, more preferably, a human.

[0147] In a preferred embodiment, the use as a medicament is for use in the prevention, amelioration and / or treatment of an acne-related disease and / or an inflammatory disease.

[0148] In a preferred embodiment, the use as a medicament is for the prevention, amelioration and / or treatment of an inflammatory disease. Also preferably, the inflammatory disease is rosacea, atopic skin or psoriasis.

[0149] In a seventh aspect, the present invention refers to the use of the composition or a peptide of the present invention for the manufacture of a medicament for the prevention, amelioration and / or treatment of acne and / or an inflammatory disease. In this seventh aspect of the present invention, the composition or peptide of the present invention is administered to a subject in need of the treatment. Preferably, the subject in need of the treatment is a mammal, more preferably, a human.

[0150] In an eighth aspect, the present invention refers to a method for the prevention, amelioration and / or treatment of acne and / or other related inflammatory diseases and / or other related skin conditions, comprising administering a peptide or a composition of the present invention to a subject in need thereof. Preferably, the subject in need of the treatment is a mammal, more preferably, a human. More preferably, the administration is topic administration.

[0151] SEQUENCE LISTING FREE TEXT

[0152] SEQ ID: 3 Letters 1 Letter

[0153] SEQ ID NO: 3 Lys-Tyr-Asp-Val KYDV

[0154] SEQ ID NO: 6 Trp-His-Tyr-Glu-Arg WHYER

[0155] SEQ ID NO: 19 Arg-Trp-Ala-His-Phe-Leu RWAHFL SEQ ID NO: 20 Arg-Trp-Leu-His-Phe-Ala RWLHFA

[0156] SEQ ID NO: 21 His-Trp-Leu-Arg-Ala-Phe HWLRAF

[0157] EXAMPLES

[0158] To allow a better understanding, the present invention is described in more detail below with reference to the enclosed drawings, which are presented by way of example, and with reference to illustrative and non-limitative examples. In the figures: * means p<0.05; ** means p< 0.01 ; *** means p<0.001 .

[0159] Abbreviations: The abbreviations used in the present text for amino acids follow the 1983 IUPAC-IUB Joint Commission on Biochemical Nomenclature recommendations outlined in Eur. J. Biochem. (1984) 138:937.

[0160] For the examples: Arg, Arginine; Asn, Asparagine; Asp, aspartic acid; Boc, tert- butyloxycarbonyl; BSA, Bovine Serum Albumin; cDNA, Complementary DNA; C-terminal or C--terminus, carboxy-terminal; DAPI, 4',6-diamidino-2-phenylindole; DCM, dichloromethane; DIPCDI, N,N'-diisopropylcarbodiimide; DMEM, Dulbecco's Modified Eagle Medium; DMF, N,N-dimethylformamide; DMSO, Dimethyl sulphoxide; DNA, Deoxyribonucleic acid; equiv, equivalent; ESI-MS, electrospray ionization mass spectrometry; FBS, Fetal Bovine Serum; Fmoc, 9-fluorenylmethyloxycarbonyl; Gin, Glutamine; Glu, Glutamic acid; Gly, Glycine; His, Histidine; HOBt, 1 -hydroxybenzotriazole; HPLC, high performance liquid chromatography; lie, Isoleucine; Leu, Leucine; Lys, Lysine; MBHA, p-methylbenzhydrylamine; Me, methyl; MeCN, acetonitrile; MeOH, methanol; N- terminal or N-terminus, amino-terminal; PBS, Phosphate Borate Saline; Pbf, 2, 2, 4,6,7- pentamethyldihydrobenzofuran-5-sulfonyl; Phe, Phenylalanine; Pro, Proline; qPCR, realtime polymerase chain reaction; RIPA, Radioimmunoprecipitation assay buffer; RNA, Ribonucleic acid; RT, room temperature; RT-qPCR, Quantitative reverse transcription Polymerase Chain Reaction; Ser, Serine; TFA, trifluoroacetic acid; Thr, Threonine; TIS, triisopropylsilane; Trp, Tryptophane; Trt, triphenylmethyl or trityl; Tyr, tyrosine, Vai, Valine.

[0161] Statistical analysis: Results are reported in tables in their respective units.

[0162] Mean values are calculated as: where: p is the value of the parameter under analysis; n is the number of subjects participating in the study Percentages are calculated as follows:

[0163] % var. or

[0164] , ,

[0165] % of subjects where: ptis the value of the parameter under analysis after product application; p0is the value of the parameter under analysis before product application; n is the number of subjects participating in the study.

[0166] The mean standard error of data is calculated as: where: p is the value of the parameter under; n is the number of subjects participating in the study

[0167] Instrumental data are submitted to 2-way Student’s test t for paired data (intra-group analysis vs TO and inter-group statistical analysis Product A vs. Product B). Variations are considered statistically significant when the p value is <0.05.

[0168] Example 1. Synthesis and preparation of the peptides

[0169] Regarding the chemical synthesis procedures included in the examples, it is noted that all synthetic processes were carried out in polypropylene syringes fitted with porous polyethylene discs or Pyrex® reactors fitted with porous plates. All the reagents and solvents were synthesis quality and were used without any additional treatment. The solvents and soluble reagents were removed by suction. The Fmoc group was removed with piperidine- DMF (2:8, volume / volume, v / v) (at least 1 x1 min, 2x10 min, 5 mL / g resin) (Lloyd Williams P. et al., Chemical Approaches to the Synthesis of Peptides and Proteins, CRC, 1997, Boca Raton (Fla., USA)). Washes between stages of deprotection, coupling, and, again, deprotection, were carried out with DMF (3x1 min) and DCM (3x1 min) each time using 10 mL solvent / g resin. Coupling reactions were performed with 3 mL solvent / g resin. The control of the couplings was performed by carrying out the ninhydrin test (Kaiser E. et al., Anal. Biochem., 1970, 34: 595598). All synthetic reactions and washes were carried out at RT.

[0170] 1. 1 Obtaining Fmoc-peptides

[0171] Weights were normalized. 4.8 g (2.5 mmol) of the Fmoc-Rink-MBHA resin with a functionalization of 0.52 mmol / g were treated with piperidine-DMF according to the described general protocol known in the state of the art in order to remove the Fmoc group. 2.90 g of Fmoc-L-Phe-OH (7.5 mmol; 3 equiv) were incorporated onto the deprotected resin in the presence of DIPCDI (1.17 mL; 7.5 mmol; 3 equiv) and HOBt (1.01 g; 7.5 mmol; 3 equiv) using DMF as a solvent for one hour.

[0172] The resin was then washed as described in the general methods known in the state of the art and the deprotection treatment of the Fmoc group was repeated to couple the next amino acid. Following the previously described protocols 2.33 g of Fmoc-L-Ala-OH (7.5 mmol; 3 equiv); subsequently 4.86 g of Fmoc-L-Arg(Pbf)-OH (7.5 mmol; 3 equiv); subsequently 2.65 g of Fmoc-L-Leu-OH (7.5 mmol; 3 equiv); subsequently 3.94 g Fmoc-L-Trp(Boc)-OH (7.5 mmol; 3 equiv) and subsequently 4.64 g of Fmoc-L-His(Trt)-OH (7.5 mmol; 3 equiv) were coupled, sequentially, each coupling in the presence of 1.01 g of HOBt (7.5 mmol; 3 equiv) and 1.17 mL of DIPCDI (7.5 mmol; 3 equiv). As already noted above, between each amino acid addition step, a deprotection treatment of the Fmoc group was performed. After the synthesis, the peptide resins were washed with DCM (5 times for 3 minutes each one) and dried under vacuum.

[0173] Using the synthesis procedures mentioned above, with the required selection of amino acids, the following sequences were synthesized:

[0174] SEQ ID NO: 3 Lys-Tyr-Asp-Val

[0175] SEQ ID NO: 6 Trp-His-Tyr-Glu-Arg

[0176] SEQ ID NO: 19 Arg-Trp-Ala-His-Phe-Leu

[0177] SEQ ID NO: 20 Arg-Trp-leu-His-Phe-Ala

[0178] SEQ ID NO: 21 His-Trp-Leu-Arg-Ala-Phe

[0179] 1.2 Removal of Fmoc N-Terminal protective group of the peptides synthesized

[0180] The N-terminal Fmoc group of the peptidyl resins obtained in 2.1 above, was deprotected with 20% (volume / volume, hereinafter v / v) piperidine in DMF (1 x1 min+2xio min) (Lloyd Williams P. et al. (1997) Chemical Approaches to the Synthesis of Peptides and Proteins. CRC, Boca Raton (Fla., USA)). The peptidyl resins were washed with DMF (5x1 min), DCM (4x1 min), and dried under vacuum.

[0181] 1.3 Cleavage process from the polymeric support of the peptidyl resins

[0182] Weights were normalized. 200 mg of the dried peptidyl resin obtained in c) were treated with 5 mL of TFA / TIS / H2O (90:5:5, in volume) for 2 hours at room temperature under stirring. The filtrates were collected and precipitated using 50 mL (8 to 10-fold) of cold diethyl ether. The ethereal solutions were evaporated to dryness at reduced pressure and room temperature, the precipitates were redissolved in 50% (v / v) MeCN in H2O and lyophilized.

[0183] 1.- 4 Characterization of the peptides synthesized

[0184] HPLC analysis of the peptides obtained in accordance with d) was carried out with a Shimadzu equipment (Kyoto, Japan) using a reverse-phase column (150x4.6 mm, XBridge Peptide BEH C18, 3.5 pm, Waters, USA) in gradients of MeCN (+0.036% (v / v) TFA) in H2O (+0.045% (v / v) TFA) at a flow rate of 1 .25 mL / min and detection was carried out at 220 nm. All peptides showed a purity exceeding 80%. The identity of the peptides obtained was confirmed by ESI-MS in a Water ZQ 4000 detector using MeOH as the mobile phase and a flow rate of 0.2 mL / min. Results obtained demonstrated that all peptides were correctly and effectively synthesized.

[0185] Additionally, the cytotoxic potential of different concentrations of the peptides (0.001 , 0.005, 0.01 , 0.05, 0.1 , 0.5 and 1 mg / mL) was determined in two different skin cell types in vitro: Human Epidermal Keratinocytes (HEKa) and Human Dermal Fibroblasts (HDFa) for 24 hours. Results showed that treatment with the peptides has no significant toxicity at 0.5 mg / mL concentration or lower, as viability remain above 80% in all concentrations tested for all the peptides (data not shown).

[0186] Example 2. Inhibition of the activity of ACC, IGF1R and MMP1

[0187] 2. 1 1nhibition ACC

[0188] To determine the effect of the peptides on the activity of ACC (Acetyl-coenzyme A (CoA) carboxylase) enzyme, it was evaluated the inhibitory effect of the peptides on ACC activity.

[0189] BPS Bioscience ACC1 Assay Kit (BPS Bioscience) to screen ACC inhibitors was used. This assay is based in the capacity of ACC to produce ADP when catalyzing the transformation of hydrogen carbonate and Acetyl-CoA to Malonyl-CoA. Candidates that inhibit the enzymatic activity of the carboxylase result in less ADP production which was measured with the ADP-Glo kit from Promega. ACC, in the presence of Acetyl-CoA, sodium bicarbonate and ATP, was exposed to different concentrations of the peptides SEQ ID NO: 21 (0.05 and 0.5 mg / mL, Fig. 1A), SEQ ID NO: 20 (0.05 and 0.1 mg / mL, Fig. 1 B), SEQ ID NO: 6 (0.1 and 0.5 mg / mL, Fig. 1C), SEQ ID NO: 19 (0.5 mg / mL) and SEQ ID NO: 3 (0.5 mg / mL, Fig. 1 D) for 40 min at room temperature. Then ADP-Glo was added to each well during 45 min. Finally, kinase detection reagent was added and incubated for at least 45 min more. Finally, luminescence of samples was measured using a microplate reader (FLUOstar Fluorimeter (BMG LabTech)). Luminescence detected is directly proportional to the ACC activity. Absorbance values were normalized using the data obtained for non-treated wells (Basal), obtaining the % of activity shown in Figure 1.

[0190] As it can be seen, the peptides inhibit ACC activity in a concentration dependent manner, reaching up to 23% inhibition with the highest concentration tested (0.5 mg / mL). This suggests that all these active ingredients can be used for anti-acne cosmetic products development.

[0191] 1.2 Inhibition MMP1

[0192] MMPs, produced by different types of cells, including keratinocytes and sebocytes, play important roles in acne inflammation, dermal matrix destruction and hyperproliferative skin disorders. MMPs also cause rupture of the pilosebaceous follicle to exacerbate inflammation. They are highly upregulated in many inflammatory states, including acne. As MMPs selectively cleave collagen, the increase in MMPs in acne is believed to be a factor in acne scarring. Therefore, ingredients that are able to decrease MMP1 activity can be of great interest for anti-acne cosmetic products development (Kang et al., 2005. Inflammation and extracellular matrix degradation mediated by activated transcription factors nuclear factor-kappaB and activator protein-1 in inflammatory acne lesions in vivo. Am J Pathol. Jun;166(6):1691-9; Papakonstantinou et al., 2005. Matrix metalloproteinases of epithelial origin in facial sebum of patients with acne and their regulation by isotretinoin. J Invest Dermatol. 2005 Oct;125(4):673-84).

[0193] In this study it was evaluated the inhibitory effect of the peptides SEQ ID NO:3, SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21 on MMP1 activity. For this purpose, it was used Abeam MMP1 Inhibitor Screening Assay Kit (Colorimetric, Abeam) that is designed to screen MMP1 inhibitors using a thiopeptide as a chromogenic substrate (Ac-PLG-[2- mercapto-4-methyl-pentanoyl]-LG-OC2H5). The MMP cleavage site peptide bond is replaced by a thioester bond in the thiopeptide. Hydrolysis of this bond by MMP1 produces a sulfhydryl group, which reacts with DTNB [5,5’-dithiobis(2-nitrobenzoic acid), Ellman’s reagent] to form 2-nitro-5-thiobenzoic acid, which can be detected by its absorbance at 412 nm. Consequently, by adding a MMP1 inhibitor agent in the mixture there would decrease the quantity of thiopeptide cleaved, and therefore, a decrease in absorbance can be observed.

[0194] 15.3 II of MMP1 were exposed to different peptide concentrations (0.01 , 0.05 and 0.1 mg / mL for SEQ ID NO:21 (Figure 2A), 0.1 and 1 mg / mL for SEQ ID NO: 3 (Figure 2B), SEQ ID NO: 19 (Figure 2C) and SEQ ID NO: 20 (Figure 2D) for 1 h at 37 °C. After exposure, the substrate was added to the mixture and the inhibitory activity was measured by spectrophotometry after 2 minutes of reaction. MMP1 activity assay was performed according to manufacturer protocol.

[0195] Each condition was performed at least in triplicate (n=3) in three independent experiments (N=3). For all data mean values of the different experiments and SEM values are shown. All data was normalized as follows:

[0196] Statistical data analysis was performed using Student t-test by comparison of % activity from test substance wells vs. non-treated wells, ***p<0.001 **p<0.01 , *p<0.05.

[0197] Results in Figure 2 show that all peptides significantly decrease MMP1 activity in a concentration dependent manner, reaching a 32% decrease with the highest concentration tested). MMP1 activity is significantly inhibited by the peptides in a dose-response manner, suggesting that this active ingredient can be used for anti-aging and antiacne cosmetic products development.

[0198] 2.3 Inhibition IGF1R

[0199] HaCaT cells were seeded in a 24-well plate at a density of 150000 cells / well and maintained at standard culture conditions (DMEM with 10% FBS and 1 % Penicillin / Streptomycin; 37°C, 95% Relative Humidity, 5% CO2) for 24 hours. Then, HaCaT were deprived for 24 hours. Next, cells were treated for 6 hours with or without the presence of 0.005, 00.1 , 0.05, 0.1 or 0.5 mg / mL of SEQ ID NO: 21 before the addition of 100 ng / mL IGF for 5 minutes. Finally, cells were lysed with RIPA buffer and lysates were collected and centrifuged in order to eliminate cell debris and were kept at -80°C until they were used for quantification of Phospho-IGF-1 R with Human Phospho-IGF-1 R / IGF1 R according to manufacturer instructions.

[0200] Figure 3 shows the effect of the different concentrations of the peptide in the inhibition of IGFI R at six hours, normalized with BCA.

[0201] Example 3. Sebocyte analysis

[0202] 3. 1 Lipid content (Bodipy)

[0203] The sebaceous gland is integral to the structure and function of the skin, providing 90% of its surface lipids. It is mainly composed by sebocytes, a highly specialized, sebumproducing epithelial cells that gradually accumulate lipids and eventually disrupt, releasing their content (sebum) by rupture of the cell membrane and cellular degradation (holocrine secretion). Sebaceous glands are most commonly in association with hair follicles (Schneider and Paus, 2010. Sebocytes, multifaceted epithelial cells: lipid production and holocrine secretion. Int J Biochem Cell Biol. Feb;42(2):181-5; Shamloul & Khachemoune, 2021. An updated review of the sebaceous gland and its role in health and diseases Part 1 : Embryology, evolution, structure, and function of sebaceous glands. Dermatol Ther. Jan;34(1): e14695).

[0204] Sebum is a light yellow and viscous secretion produced by sebaceous glands. It is an integral component of the epidermal barrier and the skin immune system, but excessive sebum production is seen in acne vulgaris, one of the most common skin diseases. This increased synthesis of sebum is highly dependent on sebocyte de novo lipogenesis (DNL), and flux through this pathway is elevated in patients with this acne. The suppression of sebum production targeting DNL is known to reduce acne incidence and severity (Esler et al., 2019. Human sebum requires de novo lipogenesis, which is increased in acne vulgaris and suppressed by acetyl-CoA carboxylase inhibition. Sci Transl Med. May 15;11 (492): eaau8465).

[0205] As sebocytes mature, they increase their size, accumulate lipid droplets in their cytoplasm and suffer nuclear degeneration (Zhang et al., 2018. Differentiation Model Establishment and Differentiation-Related Protein Screening in Primary Cultured Human Sebocytes. Biomed Res Int. Apr 5; 2018:7174561 ). Therefore, staining of lipid droplets accumulation is a good method to evaluate sebocyte differentiation stage. In this example it was evaluated the effect of peptide SEQ ID NO:21 on lipid levels of Human iPCs-derived sebocytes. For this purpose, lipid droplets will be stained using BODIPY, a well-known fluorescent probe.

[0206] Before seeding, 96-wells black plates were coated with fibronectin (Sigma) diluted 1 / 100 in PBS for at least 2 hours at 37 °C. Then, sebocytes were seeded in Sebocytes basal medium with supplement A (1 :1000) (Phenocell), at a density of 15.000 cell / well and incubated at 37°C in a humidified atmosphere with 5% CO2 for 4 days.

[0207] Next, cells were treated with SEQ ID NO: 21 at 0.01 , 0.05 or 0.1 mg / mL diluted in Sebocytes basal medium with supplement A (1 : 1000) for 24 hours more. After treatment, samples were washed three times with PBS and stained with 2 pM BODIPY 500 / 510 for 15 min. Finally, the samples were fixed with 4% formaldehyde for 10 minutes, washed three times with PBS and fluorescence images were acquired using 20x objective (Excitation: 495 / Emission: 519) in an Eclipse TS2 Microscope (Nikon). Staining was performed on at least three replicates for each condition and images of at least five different fields from each well were acquired using the same settings.

[0208] Images were analyzed using Image J software. Shortly, threshold was adjusted to select BODIPY staining and integrated density was measured, to determine fluorescence of samples. To normalize this value by the number of cells, threshold was adjusted to show the whole cell morphology. Next, threshold was applied to the pictures and watershed Imaged option was applied to segment cells. Then, analyze particles higher than 1000 px was used to determine the number of cells per image. Integrated density of each image was divided by the number of cells to obtain the fluorescence of each sample. Finally, all data was normalized as follows to obtain the relative BODIPY staining per cell in each sample compared to untreated samples (Basal): 100

[0209] Statistical data analysis was performed using Student t-test by comparison of % fluorescence from ingredient treated samples vs. untreated samples (*, p < 0.01).

[0210] Figure 4 and Table 1 show the BODIPY relative levels per cell, showing that treatment with the peptide SEQ ID NO: 21 reduces lipid levels in a concentration dependent manner, reaching up to 41% decrease after treatment with 0.1 mg / mL of the peptide.

[0211] Table 1. Lipid concentration decrease after incubation with the peptide.

[0212] Treatment of sebocytes with SEQ ID NO: 21 results into a decrease of lipid synthesis. These results suggest a potential role of SEQ ID NO: 21 to reduce sebum production, with potential cosmetic applications.

[0213] 3.2 Sebocyte size (pan-keratin)

[0214] As sebocytes mature, they increase their size, accumulate lipid droplets in their cytoplasm and suffer nuclear degeneration (Zhang et al., 2018. Differentiation Model Establishment and Differentiation-Related Protein Screening in Primary Cultured Human Sebocytes. Biomed Res Int. Apr 5; 7174561). In addition, accumulation of different cytokeratins (such as 4, 7, 13 and 19) occurs as sebocytes differentiate (Xia et al., 2009. Culture of human sebocytes in vitro. Dermatoendocrinol. Mar;1 (2):92-5). Therefore, cytokeratin accumulation is a well-known marker of sebocyte differentiation and size. In this example it was evaluated the effect of peptide SEQ ID NO: 21 on size Human iPCs-derived sebocytes. For this purpose, cytokeratins were immunostained using a pan-keratin antibody.

[0215] Before seeding, coverslips were placed in 24 wells plates and coated with fibronectin (Sigma) diluted 1 / 100 in PBS (Life Technologies) for at least 2 hours at 37°C. Then, sebocytes were seeded in Sebocytes basal medium with supplement A (1 :1000) (Phenocell) at a density of 40000 cell / well and incubated at 37°C in a humidified atmosphere with 5% CO2 for 4 days. Next, cells were treated with peptide SEQ ID NO:21 at 0.01 or 0.05 mg / mL diluted in Sebocytes basal medium with supplement A (1 :1000) for 48 h more. After treatment, samples were fixed with 4% formaldehyde and washed three times with PBS. Then, samples were blocked and permeabilized for 1 hour with PBS / 0.2% Triton X-100 / 5% BSA, in order to avoid nonspecific antibody binding and improve antibody permeation.

[0216] Next, samples were incubated overnight at 4°C with pan-keratin antibody (Proteintech) diluted 1 :200 in PBS / 0.2% Triton X- 100 / 1 % BSA. After proper washing with PBS, cells were incubated with 5 pg / mL of the secondary antibody Goat anti-Rabbit IgG Alexa Fluor 488 (recognition of primary antibody) (Thermofisher) and 0.4X Phalloidin-TRITC (for actin filaments staining) (Sigma) during 1 hour at room temperature and dark condition. Finally, samples were washed three times with PBS and coverslips were mounted using Prolong-Gold with DAPI (Thermofisher), which strongly binds DNA (nuclei staining), and samples were kept protected from light at 4°C until microscopic images were acquired.

[0217] Microscopic images were acquired using 20x objective in an Eclipse TS2 Microscope (Nikon). Immunofluorescence was performed on at least three replicates for each condition and images of at least five different fields from each coverslip were acquired using the same settings.

[0218] Images were analyzed using Image J software. Threshold was adjusted to select pankeratin staining and stained area was measured. The number of cells was counted using the DAPI staining. Pan-keratin-stained area was divided by the number of cells. Finally, all data was normalized as follows to obtain the % of pan-keratin compared to untreated cells (Basal):

[0219] Statistical data analysis was performed using Student t-test by comparison of % fluorescence from ingredient treated samples vs. untreated samples, (*, p < 0.05).

[0220] Figure 5 and Table 2 show that sebocytes treated with SEQ ID NO: 21 reduced pan-keratin staining in a concentration dependent manner, reaching up to 51 % decrease after treatment with 0.05 mg / mL of the ingredient.

[0221] Table 2. Data of the analysis results shown in Figure 5.

[0222] Treatment of sebocytes with SEQ ID NO:21 resulted into a decrease of pan-keratin-stained area. Given that pan-keratin levels are correlated with sebocyte size, it can be concluded that the peptide reduces cell size. These results suggest a potential role of SEQ ID NO: 21 to reduce sebum production, with potential cosmetic applications.

[0223] 3.3 Potential of the peptides modulating the expression of sebum-related genes in Human iPSCs-derived sebocytes.

[0224] The potential of the peptide SEQ ID NO: 21 to modulate the expression of genes involved in sebocytes maturation and sebum production in human sebocytes was analyzed.

[0225] Briefly, early sebocytes were seeded in triplicate (n=3) in 6-well plates at a density of 3.5-4 x 105cells / well and maintained at standard culture conditions (Sebocytes basal medium with supplement A (1 :1000) (Phenocell); 37°C, 5% CO2) for 4 days. Then, cells were treated 6 or 24 hours more with basal medium supplemented with supplement A (1 :1000) alone or in the presence of 0.01 or 0.005 mg / mL SEQ ID NO: 21 .

[0226] Finally, cells were lysed, and replicates were pooled together for RNA extraction using Qiagen RNeasy Mini kit following manufacturer’s instructions. Purified RNAs were used to generate the corresponding cDNAs by reverse transcription using a commercial kit (High- capacity cDNA reverse transcription kit, Applied Biosystems) which served as templates for amplification. RT-qPCR was performed with the panel of TaqMan assay probes specified above and 2x gene expression Master Mix using StepOne plus Real-Time PCR instrument. Amplification was performed following described methodology (Arya et al., 2005. Basic principles of real-time quantitative PCR, Expert Rev. Mol. Diagn., 5(2):209-19; Jozefczuk et al., 2011. Quantitative real-time PCR-based analysis of gene expression. Methods Enzymol. 500:99-109).

[0227] The obtained data were analyzed using the AACt method, which provides the target gene expression values as fold changes in the treated sample compared with an untreated basal sample. Both samples were normalized with the relative expression of a housekeeping gene: GAPDH (Glyceraldehyde 3-phosphate dehydrogenase).

[0228] The steps for analysis include:

[0229] 1 . Calculate the average Ct for each condition

[0230] 2. Calculate the ACT test sample and the ACT calibrator sample

[0231] 3. Calculate the AACT: AACT = ACT test sample - ACT calibrator sample

[0232] 4. Obtain ratio by 2'AACT

[0233] Gene modulations with a ratio (fold change vs. basal) >1.2 or < -1.2 are considered significant and of potential biological importance. Results showed that after treatments with SEQ ID NO: 21 , upregulation of FOXO1 gene can be observed at 6 h of treatment, whereas MLIC1 , IGF1 R, ACACA, ACACB, NR1 H3, PTGS2 and TLR2 are downregulated (Figure 6). Table 3. Mean normalized expression and fold change vs. Basal for each gene analyzed in sebocytes treated with 0.01 or 0.005 mg / mL SEQ ID NO: 21 for 6 or 24 h.

[0234] As may be seen, peptide SEQ ID NO: 21 modulates the expression of several genes involved in sebocytes maturation or sebum synthesis. These results suggest that the peptide could have an effect reducing sebum production, with potential cosmetic applications.

[0235] 3.4 Analysis of the effect of peptides on NR1H3 levels in human iP-SCS-derived sebocytes NR1 H3 (nuclear receptor subfamily 1 group H member 3, also known as Liver X Receptorci, LXR-a) is a key element in sebum production, as its activation induces lipid synthesis. In addition, it induces the expression of Sterol Regulatory Element-Binding Protein 1 (SREBP- 1) that regulates genes required for fatty acid and lipid metabolism and production (Bakry et al., 2017. Immunohistochemical Expression of Cyclo-oxygenase 2 and Liver X Receptorci in Acne Vulgaris. J Clin Diagn Res. Sep; 11 (9): WC01-WC07).

[0236] NR1 H3 plays vital roles in lipid metabolism and cholesterol homeostasis, acting as a cholesterol sensor. In the human epidermis, cholesterol contributes to the formation of the permeability barrier via lamellar body formation and its levels are significantly higher in acne patients (El-Akawi et al., 2007. The relationship between blood lipids profile and acne. J Health Sci. 53:596-99).

[0237] In this study, it was evaluated the effect of the peptide SEQ ID NO: 21 on NR1 H3 levels in Human iPCs-derived sebocytes (hereafter, sebocytes) through fluorescence immunostaining.

[0238] Before seeding, coverslips were placed in 24 wells plates and coated with fibronectin (Sigma) diluted 1 / 100 in PBS (Life Technologies) for at least 2 hours at 37°C. Then, sebocytes were seeded in Sebocytes basal medium with supplement A (1 :1000) (Phenocell), at a density of 60 000 cell / well and incubated at 37°C in a humidified atmosphere with 5% CO2 for 4 days.

[0239] Next, cells were treated with SEQ ID NO: 21 at 0.005 mg / mL diluted in Sebocytes basal medium with supplement A (1 :1000) for 24 h more. After treatment, samples were fixed with 4% formaldehyde (Sigma) and washed three times with PBS. Then, samples were blocked and permeabilized for 1 hour with PBS / 0.2% Triton X-100 / 5% BSA (Sigma), in order to avoid nonspecific antibody binding and improve antibody permeation.

[0240] Samples were incubated overnight at 4°C with NR1 H3 antibody (Proteintech) diluted 1 :200 in PBS / 0.2% Triton X-100 / 1 % BSA. After proper washing with PBS, cells were incubated with 5 pg / mL of the secondary antibody Goat anti-Rabbit IgG Alexa Fluor 488 (Thermofisher) during 1 hour at room temperature and dark conditions.

[0241] Finally, samples were washed three times with PBS and coverslips were mounted using Prolong-Gold with DAPI (Thermofisher), which strongly binds DNA (nuclei staining), and samples were kept protected from light at 4°C until microscopic images were acquired.

[0242] Microscopic images were acquired using 20x objective in an Eclipse TS2 Microscope (Nikon). Immunofluorescence was performed on at least three replicates for each condition and images of at least five different fields from each coverslip were acquired using the same settings.

[0243] Images were analysed using Image J software. Threshold was adjusted to select NR1 H3 staining and mean fluorescent was measured. The number of cells was counted using the DAPI staining. NR1 H3 mean fluorescent intensity was divided by the number of cells. Finally, all data was normalized as follows to obtain the % of NR1 H3 compared to untreated cells (Basal).

[0244] Statistical data analysis was performed using Student t-test by comparison of % fluorescence from ingredient treated samples vs. untreated samples, (*, p < 0.05).

[0245] Figure 7 shows that sebocytes treated with SEQ ID NO: 21 at 0.005 mg / mL for 24 h showed a 54% reduction of NR1 H3 (See the specific values in Table 4).

[0246] Table 4. Data of the analysis of results shown in Figure 7.

[0247] Treatment of sebocytes with SEQ ID NO: 21 results into a decrease of NR1 H3 staining. Given that NH1 R3 levels are correlated with sebocyte lipid content and pathogenesis of acne, these results suggest a potential role of SEQ ID NO: 21 to reduce sebum production, with potential anti-acne applications.

[0248] Example 4. Assays in human epidermal keratinocytes

[0249] 4. 1 Analysis of the effect of the peptides on the expression of acne-related genes in human epidermal keratinocytes

[0250] It was analyzed the potential of the peptides to modulate the expression of several acne- related genes in human epidermal keratinocytes from adult (HEKa) in vitro.

[0251] HEKa were seeded in duplicate (n=2) in 12-well plates at a concentration of 150 000 cells / well in Dermal Cell Basal Medium (DCBM). Then, cells were incubated at 37°C in a humidified atmosphere with 5% CO2. Next, cells were treated with 0.005 mg / mL SEQ ID NO: 21 and / or 107CFUs (Colony forming Units) of C. acnes for 24 hours. Finally, cells were lysed and replicates were pooled together for RNA extraction using RNeasy Mini kit (Qiagen) following manufacturer’s instructions. Purified RNAs were used to generate the corresponding cDNAs by reverse transcription using a commercial kit (see Example 4.3).

[0252] The obtained data were analyzed using the AACt method (see Example 4.3), which provides the target gene expression values as fold changes in the treated sample compared with an untreated basal sample. Gene modulations with a ratio (fold change vs. basal) >1.2 or < -1.2 are considered significant and of potential biological importance.

[0253] After a 24-hour treatment with 0.005 mg / mL SEQ ID NO: 21 , upregulation of GATA6 was observed, whereas KRT16, MMP1 , IL1 B, ACACB, ACACA, IL1 A, IGF1 R, TNF, TLR2 and MMP9 were downregulated (Figure 8). In addition, cells treated with 107CFUs of C. acnes showed upregulation of MMP1 , IL1 A, IL1 B and CXCL8 genes, whereas the presence of 0.005 mg / mL SEQ ID NO: 21 during C. acnes treatment can partially or totally revert this effect (Figure 9).

[0254] Table 5: Mean normalized expression and fold change vs. Basal for each gene analyzed in HEKa treated with 0.005 mg / mL SEQ ID NO: 21 for 24 h.

[0255] Table 6. Mean normalized expression and fold change vs. Basal for each gene analyzed in HEKa treated with 107CFUs of C. acnes alone or in the presence of 0.005 mg / mL SEQ ID NO: 21 for 24 h.

[0256] Peptide SEQ ID NO:21 modulates the expression of several genes related with acne. Moreover, SEQ ID NO:21 can revert the effect of C. acnes over several acne-related genes. All these results strongly suggest that SEQ ID NO:21 can be used for the development of cosmetic products with antiacne effect.

[0257] 4.2 Inflammation assay with C. acnes

[0258] The effect of SEQ ID NO: 21 on the release of interleukin 1 alpha (IL1 a) induced by C. acnes was determined in human epidermal keratinocytes from adult (HEKa) through enzyme linked immunosorbent assay (ELISA). Briefly, HEKa cells were seeded in a 96-well plate at a density of 18.000 cells / well and maintained at standard culture conditions (Supplemented Dermal Cell Basal Medium, DCBM; 37°C, 95% Relative Humidity, 5% CO2) for 24 hours. Then, cells were deprived for 24 h in DCBM supplemented 1 :10. Next, cells were treated for 48 hours with 107colony forming units (CFLIs) of C. acnes alone or in the presence of SEQ ID NO:21 (0.01 or 0.05 mg / mL) diluted in DCBM supplemented 1 :10.

[0259] After treatment, supernatants were collected and centrifuged to eliminate cell debris and were kept at -80°C until they were used for IL1a quantification with a Human IL1a / IL 1 F1 DuoSet ELISA kit according to manufacturer instructions.

[0260] In order to determine cell viability at the end of the experiment, 200 pL of Alamar Blue, diluted 1 :10 in complete medium, were added to each well. Cells were incubated for 2 h and then Alamar Blue was transferred to a 96-well black plate.

[0261] Finally, fluorescence of the stored 96-well black plate with Alamar Blue solutions was measured using FLUOstar Fluorimeter (Excitation: 530 nm; Emission: 590 nm) to establish cell viability.

[0262] Each condition was performed at least in triplicate (n=3) in three independent experiments (N=3). For all data, mean values of the different experiments and SEM values are shown. All data was normalised as follows:

[0263] ILla released per well

[0264] ILla released / cell = — — — - — - — — - -

[0265] RFUs Alamar Blue (590 nm)

[0266] ILla per cell in treated samples

[0267] % vs Basal = — - — — - - - - — x 100

[0268] ILla per cell in C. acnes treated samples

[0269] Statistical data analysis was performed using Student t-test by comparison of % I L1 a protein release from positive control cells (C. acnes alone) vs. untreated ones (Basal) or the other experimental conditions *p<0.05, **p<0.01 , ***p<0.001.

[0270] Figure 10 shows the results of this experiment, where treatment with C. acnes significantly increases I L1 a levels by 41%. By contrast, the presence of SEQ ID NO:21 during C. acnes treatments lowered IL1 a levels, showing up to 19% decrease at the highest concentration tested (0.05 mg / mL SEQ ID NO:21), when compared with cells treated with C. acnes alone (see Table 7 for specific values). Table 7. Data of the analysis of results shown in Figure 10.

[0271] As previously seen, peptide SEQ ID NO:21 presents an anti-inflammatory efficacy. Moreover, SEQ ID NO:21 can revert the effect of C. acnes over IL1a. All these results strongly suggest that SEQ ID NO:21 can be used for the development of cosmetic products with antiacne effect.

[0272] 4.3 Analysis of the effect of the peptides on the number of Ki67 positive human epidermal keratinocytes

[0273] Hyperkeratinization occurs when the cells of the follicle become cohesive and do not shed normally onto the skin’s surface. The result is a microcomedone (the precursor to acne) and subsequent lesions characteristic of acne (Kurokawa and Nakase, 2020. Recent advances in understanding and managing acne. F1000Res. Jul 29; 9:F1000 Faculty Rev-792). Regarding bacterial colonization and inflammatory response, the skin commensal C. acnes is thought to trigger an inflammatory response and lead to subclinical and inflammatory acne lesions. The mechanisms proposed for C. acnes pathogenesis include changes in sebaceous gland activity, comedone formation (through hyperkeratinization), and host inflammation (Xu and Li, 2019. Acne, the Skin Microbiome, and Antibiotic Treatment. Am J Clin Dermatol. Jun;20(3):335-344).

[0274] Ki67 is an antigen expressed in all stages of the cell cycle except Go. It is a nuclear protein that is associated with cellular proliferation and ribosomal RNA transcription and is necessary for the maintenance of cell proliferation (Heenen et al., 1998). Therefore, it is commonly used as a cellular marker for proliferation.

[0275] In this study, it was evaluated the effect of the peptide SEQ ID NO:21 on the number of Ki67 positive cells in Human Epidermal Keratinocytes (HEKa) through immunostaining.

[0276] HEKa cells were cultured on coverslips in Dermal Cell Basal Medium (DCBM) supplemented the Keratinocyte Growth Kit (ATCC) at a density of 35.000 cell / well and incubated at 37 °C in a humidified atmosphere with 5% CO2. 24 h later, HEKa were deprived by incubation overnight with DCBM without any supplement. Then, cells were treated with 108colony forming units (CFLIs) of C. acnes alone or in the presence of SEQ ID NO:21 (0.01 or 0.05 mg / mL) for 6 hours.

[0277] After treatments, samples were fixed with 4% formaldehyde and washed three times with PBS. Then, samples were blocked and permeabilized for 1 hour with PBS / 0.2% Triton X- 100 / 5% BSA, to avoid nonspecific antibody binding and improve antibody permeation. Next, samples were incubated 90 min at room temperature with Ki67 antibody (Proteintech) diluted 1 :200 in PBS / 0.2% Triton X- 100 / 1 % BSA. After proper washing with PBS, cells were incubated with 5 pg / mL of the secondary antibody Goat anti-Rabbit IgG Alexa Fluor 488 (Thermofisher) during 1 hour at room temperature and dark conditions.

[0278] Finally, samples were washed three times with PBS and coverslips were mounted using Prolong-Gold with DAPI (Thermofisher), which strongly binds DNA (nuclei staining), and samples were kept protected from light at 4°C until microscopic images were acquired with Eclipse TS2 Microscope (Nikon). Microscopic images were acquired using 20x objective. Immunofluorescence was performed on at least three replicates for each condition and images of at least five different fields from each coverslip were acquired using the same settings.

[0279] Images were analyzed using Image J software. Shortly, threshold was adjusted to select nuclear Ki67 staining, and the number of positive nuclei was counted using “Analyze particles” tool. Then, the number of cells was counted using the DAPI staining. Next, Ki67 positive nuclei were divided by the total number of cells of the sample and multiplied by 100 to obtain the percentage of Ki67-positive cells.

[0280] Finally, all data was normalized as follows to obtain the % of Ki67 positive cells compared to C. acnes condition control. Statistical data analysis was performed using Student t-test by comparison of % of Ki67. 100

[0281] Statistical data analysis was performed using Student t-test by comparison of % of Ki67 positive nuclei from ingredient treated samples vs. C. acnes stimulated samples, (*, p < 0.05; “, p < 0.01 and ***, p < 0.001). Figure 11 shows that the presence of SEQ ID NO:21 completely counteracts this increase in a concentration-dependent manner. HEKa treated with C. acnes alone presented a 29% increase in Ki67 positive nuclei, which is related with an increase of cell proliferation. However, the presence of SEQ ID NO:21 completely reduced this increase, recovering basal levels 6 hours after treatment (See the specific values in Table 8).

[0282] Table 8. Data of the analysis of results shown in Figure 11 .

[0283] Treatment of HEKa with SEQ ID NO:21 reverts the effect of C. acnes over keratinocyte proliferation. Given that this phenomenon is related with hyperkeratinization, these results suggest a potential role of SEQ ID NO:21 to reduce acne, with potential cosmetic applications.

[0284] Example 5: Characterization of the effect of an active ingredient on the organization of the extracellular matrix around skin pores and on the tightening of skin pores

[0285] The objective was the evaluation of the effect of the peptides on the organization of the extracellular matrix around skin pores and the tightening of skin pores. These results reinforce the cosmetic use of the peptides.

[0286] An abdominal skin explant was used for this study and was received in a survival medium (DMEM-antibiotics). We had 2 conditions, and it were used 3 punches per condition. The detailed conditions were:

[0287] Untreated skin

[0288] - Active ingredient-treated skin (SEQ ID NO:21)

[0289] Adipose tissue was removed from the abdominoplasty before the experiments. From this skin explant, skin punches were cut and then placed at the air / liquid interface in a survival medium (Medium: Gibco DMEM + antibiotic cocktail). This allows the skin punches to be hydrated by the dermis. The skin punches were placed at controlled temperature and humidity during the experiment (respectively 37°C and relative humidity: > 93%). After three days of treatment, the skin punches have either been frozen in liquid nitrogen to perform cryocuts for AFM (Atomic Force Microscopy) measurements or fixed in formaldehyde for SPIM sample processing.

[0290] The Atomic Force Microscope used in this study is a Bioscope Resolve (Bruker) on which is added an epifluorescence microscope (Leica DMi8). This configuration allows the precise positioning of the AFM probe on the sample. This combination also allows to acquire correlative images from mechanical to fluorescent acquisitions.

[0291] For this study, the AFM measurements consist in the acquisition of “force-volume (FV)” around the skin pores. A force-volume consists in the acquisition of a multitude of measurement points from which the mechanical properties (elastic modulus) are extracted. The quantification of the elastic modulus (Ea) from the raw force curves is performed using BioMeca Analysis® processing software. The raw approach curves obtained for each force-volume were processed to extract a quantification of the skin Ea. The principle consists in applying a theoretical physical model (Sneddon model) to the approach curve to calculate the elastic modulus. The elastic modulus (Ea) is expressed in kilopascal (kPa) and corresponds to the stiffness of a material. The higher the Ea value, the stiffer the material is. The Ea evaluates the resistance of a material to undergo elastic deformation when a force is applied on it. A material with high Ea is described as stiff, and conversely, materials whose Ea are low are described as “soft”, “elastic” or” flexible”. Z (pm) is the distance traveled by the AFM probe; and “force” is the relative force applied. Technically, the tip approaches the sample, comes in contact, and then penetrates it to get information about its deformation properties. The theoretical model is applied on a defined indentation range, here delimited by the blue bars.

[0292] Lightsheet imaging was performed using a Zeiss Lightsheet Z.1 microscope with a X5 objective. The Phalloidin signal was acquired using an excitation wavelength of 561 nm and a band-pass emission filter using 575 - 615 nm window was used. Technically, each fixed skin punches were processed into small pieces of 2 mm3of volume and a phalloidin labelling was performed followed by several washes in 1X PBS. Then, a dehydration by several washes in alcohol was carried out and submitted to a clearing step. The labelled small pieces of skin were then attached to the holder of the Lightsheet microscope to locate and image the skin pores. The stiffness of the dermis was first measured by AFM around the pores to determine whether the active ingredient could induce a reorganization of the dermal matrix (i.e. extracellular matrix, including collagen network). Results of the collagen stiffness around the skin pores by AFM analysis are presented in Figure 12. The table with the data is presented in Table 9.

[0293] Table 9: Collagen stiffness around the skin pores by AFM analysis.

[0294] The result from the AFM analysis of the stiffness of the collagen network surrounding the skin pores revealed a significant increase of the stiffness after the skin treatment compared to the untreated skin by 8.14 points. This increase of stiffness indicates an increase of the firmness of the dermis around the skin pores and therefore, a restructuring of the matrix around the skin pores.

[0295] It was then evaluated the diameter of the skin pores after treatment to determine if the active ingredient could also have an astringent effect. The diameter of the skin pores was assessed by SPIM imaging that allows to image large sample such as skin punches. Results of the diameter of the pores by the SPIM analysis are presented in Figure 13 and Table 10.

[0296] Table 10. Diameter of the skin pores

[0297] The size of the skin pores was assessed by Lightsheet imaging. The results after treatment overall indicate that the average diameter of the skin pores significantly decreases by 8.39 points at depth 150 pm after three days of treatment by the peptide compared to the control. In addition, a decrease in the average diameter of the skin pores is also observed at Depth 500 pm. The peptide tends to decrease the average diameter of the skin pores by 15.1 points compared to the control.

[0298] The peptide significantly increases the stiffness of the matrix around the skin pores and significantly reduces the diameter of the skin pores. Consequently, the peptide can modify the biomechanical properties of the dermal matrix around pores.

[0299] Example 6: Evaluation of the efficacy of a cosmetic product for a face care

[0300] The next step was the evaluation of the efficacy of the peptides as a cosmetic product for face care in improving acne prone skin.

[0301] In order to reach this goal a clinical instrumental study is carried out on 40 healthy Caucasian female subjects, aged between 18 and 35 years old, with mild to moderate acne on face according to GEA SCALE (level 1-2-3). According to a previously defined randomization list the subjects are divided in two groups: 20 subjects apply the active product and 20 the placebo formulation. The study foresees 28 days of use: evaluations are performed at baseline (TO), after 7 (T7), 14 (T14) and 28 days (T28) of use. Evaluations are carried out by means of non-invasive bioengineering techniques able to measure skin sebum content, skin profilometry (pores diameter) and skin moisturization. The analysis are integrated with image analysis for protoporphyrin distribution, expert scoring for lesions count, only at T28 biochemical analysis of the protection from oxidative stress (MDA test) and a self-assessment questionnaires filled in by the enrolled subjects at each checkpoint.

[0302] The products were applied in whole face, twice daily, morning and before bedtime with circular motions. The tested cosmetic products conform to Regulation (EC) No 1223 / 2009 of the European Parliament and of the Council of 30 November 2009 on cosmetic products (recast) (Text with EEA relevance) and to its annexes.

[0303] Product B comprises SEQ ID NO: 21 as active ingredient in a media, whereas product A is only the media (water (EAll), isononyl / isononanoate, caprylic / capric triglyceride polyglyceryl-4 diisostearae / polyhydroxystearate / sebacate, synthetic beeswax, glycerin, nylon-12, dimethicone, magnesium sulfate, caprylyl glycol, dimethicone / vinyl dimethicone crosspolymer, triethyl citrate, disodium EDTA, benzoic acid).

[0304] 6. 1 Skin sebum content

[0305] The sebum measurement is based on the internationally recognized Sebumeter® method (Sebumeter 815, Courage+Khazaka GmbH). The measurement principle is the photometric method, the grease spot photometer. According to this method the sebum on skin is collected on a 64 mm2 mat synthetic tape contained in a cassette, then the measuring head of the cassette is inserted into the aperture of the device, where a photocell measures the transparency. The light transmission represents the sebum content on the surface of the measuring area. A microprocessor calculates the result, which is shown on the display in pg sebum / cm2of the skin.

[0306] Figure 13 shows the results of the assay. The upper graph (A) shows the mean data obtained at each experimental time for the analyzed skin parameter. The data are expressed as a mean ± SE. The graph below (B) reports the mean % variations obtained at each timepoint for the analyzed skin parameter. Black bars correspond to the sample without the active ingredient (placebo), whereas white bars are the control with the peptide. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO). The above asterisks in (A): * p<0.05, ** p<0.01 , *** p<0.001 inter-group statistical analysis (Product A vs Product B, carried out on % variations).

[0307] Product A (placebo) determines a statistically significant decrease vs TO of skin sebum content after 28 days of product use by -7.7%. On the other side, the composition with the peptide determines a statistically significant decrease vs TO of skin sebum content after 7, 14 and 28 days of product use respectively by -11 .3%, by -15.4% and by -20.2%. Moreover, results obtained with Product B are significantly greater compared to those obtained with Product A, at each checkpoint.

[0308] 6.2 Pores size

[0309] Skin surface is quantitatively assessed by PrimosCR SF (Canfield Scientific Europe, BV, Utrecht, Netherlands). PrimosCR SF is a non-contact in vivo skin measurement device based on structured light projection. In conjunction with a comprehensive 3-D measurement and evaluation software, the sensor allows to evaluate skin surface properties (i.e. wrinkle depth, volume, roughness etc.). In this study pores size is evaluated.

[0310] Figure 14 shows the mean data obtained at each experimental time for the analyzed skin parameter (A). The data are expressed as a mean ± SE. * p<0.05, **p<0.01 , ***p<0,001 intragroup statistical analysis (T7 / T14 / T28 vs TO). Upper asterisks: * p<0.05, ** p<0.01 , *** p<0.001 inter-group statistical analysis (Product A vs Product B, carried out on % variations). The graph below (B) reports the mean % variations obtained at each timepoint for the analyzed skin parameter.

[0311] No significant variations of the monitored parameter are observed with the use of without peptide, Product A (black bars) at each checkpoint. The product with the active ingredient (Product B, white bars) determines a statistically significant reduction vs TO of pores size after 7, 14 and 28 days of product use respectively by -2.8%, by -5.0% and by -8.0%. Moreover, results obtained with Product B are significantly greater compared to those obtained with Product A, at T14 and T28.

[0312] 6.3 Lesion count - parameter evaluated all over the face

[0313] The experimenter visually assessed (and by palpation if necessary) the facial skin, counting the inflammatory (papules and pustules) and non-inflammatory (comedones: blackheads and whiteheads) lesions. The lesions count is mainly based on the evaluation of the total number of lesions on the face of the subject.

[0314] It was counted the number of acne lesions (both inflammatory and non-inflammatory) at each timepoint for each volunteer. Data are expressed as lesion number. Figures 15A to 15D reports the mean data obtained at each experimental time for the analyzed skin parameter. The data are expressed as a mean ± SE.

[0315] The composition comprising the peptide (White bars) determines a statistically significant decrease vs TO of the total number of acne lesions (inflammatory and non-inflammatory) after 7, 14 and 28 days of product use respectively by -1.8 (mean percentage variation by - 8.7%), by -2.2 (mean percentage variation by -10.1 %), and by -3.1 (mean percentage variation by -15.0%).

[0316] 6.4 Analysis of protoporphyrin distribution - parameter evaluated all over the face

[0317] On pictures acquired by Visia®-CR (Canfield Scientific) an image analysis with a dedicated software is carried out in order to evaluate Protoporphyrin distribution. Protoporphyrin is produced by bacteria and a decrease of its distribution can be related to impurities removal.

[0318] Figure 16 shows the mean data obtained at each experimental time for the total area occupied by protoporphyrin (a.u.). The graph below reports the analyzed skin parameter. The data are expressed as a mean ± SE.

[0319] The product comprising the peptide (white bars) determines a statistically significant decrease vs TO of the total area occupied by protoporphyrin after 14 and 28 days of product use respectively by -5.3% and by -7.8%; a decrease of protoporphyrin distribution is related to impurities removal. Moreover, results obtained with Product B are significantly greater compared to those obtained with without the peptide, at T28.

[0320] 6.5 Skin moisturization - parameter evaluated on cheek

[0321] Skin moisturization was evaluated by means of Corneometer® measurement. This measurement is based on the completely different dielectric constant of water and other substances (mostly <7). The measuring capacitor shows changes of capacitance according to the moisture content of the skin. A glass lamina separates the metallic tracks (gold) in the probe head from the skin in order to prevent current conduction in the measured area. An electric field between the tracks with alternating attraction develops. One track builds up a surplus of electrons (minus charge) the other a lack of electrons (plus charge). The scatterfield penetrates the very first layer of the skin (10-20 pm) during the measurement and the capacitance was determined.

[0322] Figure 17 shows the data for the skin moisturization at each timepoint for each volunteer. Data are expressed as corneometric units (c.u.), treated with the composition comprising the peptide (White bars) or the control (black bars), whereas in B the mean of % variation at each timepoint is shown.

[0323] The composition comprising the peptide determines a statistically significant increase vs TO of skin moisturization after 7, 14 and 28 days of product use respectively by +5.8%, by +8.0% and by +8.7%. Moreover, these results B are quantitively greater compared to those obtained with the control.

[0324] 6.6 Biochemical analysis of the protection from oxidative stress (MPA test) / Lipoperoxidation

[0325] Samples of the first layers of the stratum corneum were collected by the experimenter from the cleaned skin of the face using Corneofix® foils (Courage+Khazaka electronic GmbH); in particular, different layers of stratum corneum were collected from the same skin area. The first stripped layer is discarded and then 10 consecutive strips are collected and stored at -80°C upon further analysis.

[0326] Lipoperoxides are measured by means of Malondialdehyde assay (MDA). Malondialdehyde (MDA) and 4- hydroxynonenal are the two main products of lipid peroxidation. Their concentration in a biological system is a good index of its lipoperoxide damage. To determine the lipoperoxides levels it was assayed the method tested by Erdelmeier and collaborators (1998): the assay is based on the capability of a cromogen, N methyl 2 phenylindole (NMPI), to react with MDA at 45°C and acid pH to produce a stable chromophore that has an absorption peak at 586 nm. The lipoperoxide levels were measured after the induction of unstable hydroperoxides decomposition, produced in the oxidative processes by means of a pro-oxidant agent (CuSC t 500 mM).

[0327] Figure 18 shown the data of the experiment, comparing the composition with the peptide (white bars) and the composition without the peptide (control), black bars. Figure 18B shows the mean % variations between T28 and TO. It is proved that the composition comprising the peptide determines a statistically significant reduction vs TO of Lipo-PerOxide levels after 28 days of product use by -22.4%. Moreover, results obtained with this product B are significantly greater compared to those obtained with the control.

[0328] To summarize the cosmetic effects of the composition, the inclusion of the peptide produces a decrease of skin sebum content by -11.3%* at T7, by -15.4%* at T 14 and by -20.2%* at T28; a decrease of pores size (diameter) by -2.8%* at T7, by -5.0%* at T14 and by -8.0%* at T28; a decrease of the number of the total number of acne lesions (inflammatory and non-inflammatory) by -1.8* at T7, by -2.2* at T14 and by -3.1* at T28; a decrease of the total area occupied by protoporphyrin (related to impurities removal) by -5.3%* at T14 and by - 7.8%* at T28; an increase of skin moisturization by +5.8%* at T7, by 8.0%* at T14 and by +8.7%* at T28; a decrease of Lipo-PerOxide levels (measured by means of MDA assay) by -22.4%* at T28.

Claims

CLAIMS1. Peptide which interacts with ACC, MMP1 and / or IGF1 R selected from:Ri-Lys-Tyr-Asp-Val-R2(SEQ ID NO: 3);Ri-Trp-His-Tyr-Glu-Arg-R2(SEQ ID NO: 6);Ri-Arg-Trp-Ala-His-Phe-Leu-R2(SEQ ID NO: 19)Ri-Arg-Trp-leu-His-Phe-Ala-R2(SEQ ID NO: 20); or Ri-His-Trp-Leu-Arg-Ala-Phe-R2(SEQ ID NO: 21), its acceptable isomers, salts, solvates and / or mixtures thereof, wherein:Ri is selected from H, substituted or unsubstituted non-cyclic aliphatic, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl and Rs-CO-, wherein R5 is selected from the group formed by substituted or unsubstituted Ci-C24 alkyl radical, substituted or unsubstituted C2-C24 alkenyl, substituted or unsubstituted C2-C24 alkynyl, substituted or unsubstituted Cs-C24 cycloalkyl, substituted or unsubstituted Cs-C24 cycloalkenyl, substituted or unsubstituted Cs-C24 cycloalkynyl, substituted or unsubstituted C6-C30 aryl, substituted or unsubstituted C?-C24 aralkyl, substituted or unsubstituted heterocyclyl ring of 3 to 10 members, and substituted or unsubstituted heteroarylalkyl of 2 to 24 carbon atoms and 1 to 3 atoms other than carbon and an alkyl chain of 1 to 6 carbon atoms; andR2is selected from H, -NR3R4- , -OR3 and -SR3, wherein R3 and R4 are independently selected from H, substituted or unsubstituted non-cyclic aliphatic group, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, and substituted or unsubstituted aralkyl.

2. The peptide of claim 1 , wherein it is a head-to-tail cyclic peptide.

3. The peptide of claims 1 or 2, wherein it is selected from:Ri-Lys-Tyr-Asp-Val-R2(SEQ ID NO: 3)Ri-Arg-Trp-Ala-His-Phe-Leu-R2(SEQ ID NO: 19)Ri-Arg-Trp-leu-His-Phe-Ala-R2(SEQ ID NO: 20); orRi-His-Trp-Leu-Arg-Ala-Phe-R2(SEQ ID NO: 21).

4. The peptide of any of claims 1 to 3, wherein Ri is H or acetyl and R2 is NH2 or OH.

5. The peptide of claim 4, wherein the peptide is selected from: Ac-Lys-Tyr-Asp-Val-NH2(SEQ ID NO: 3) Ac-Arg-Trp-Ala-His-Phe-Leu-NH2(SEQ ID NO: 19) Ac-Arg-Trp-leu-His-Phe-Ala-NH2(SEQ ID NO: 20); or Ac-His-Trp-Leu-Arg-Ala-Phe-NH2(SEQ ID NO: 21).

6. A cosmetic composition comprising the peptide of any of claims 1 to 5.

7. A pharmaceutical composition comprising the peptide of any of claims 1 to 5.

8. Use of the cosmetic composition of claim 6 for skin pore reduction, sebum control, redness, dryness, roughness, wrinkles and / or increase of hydration of the skin in a subject.

9. The peptide of claims 1 to 5 or the pharmaceutical composition of claim 7 for use as a medicament.

10. The peptide of claims 1 to 5 for use in the reduction of inflammation of the skin.

11. The peptide of claims 1 to 5 or the pharmaceutical composition of claim 7 for use in the prevention, amelioration and / or treatment of an acne-related disease and / or an inflammatory disease.