Peptides and compositions for use in cosmetics, foods and pharmaceuticals

Abstract

The present invention relates to peptides for providing telomere protection, antioxidant and anti-inflammatory activity and thus useful in cosmetics for preventing, reducing and / or treating inflammatory aging and skin aging, and in pharmaceuticals for treating age-related diseases, inflammatory diseases and diseases related to oxidative stress, and in the food industry as dietary supplements.

Description

Detailed Description of the Invention

[0001] FIELD OF THEINVENTION The present invention relates to the field of molecular biology, more precisely to molecular biology applied to cosmetics, nutraceuticals and pharmaceuticals, and even more precisely to peptides and compositions comprising said peptides, capable in particular of protecting telomeres and protecting against inflammation and oxidation, said peptides therefore helping in the prevention and / or treatment of aging, including inflammation, preferably aging of the skin.

[0002] BACKGROUND OF THEINVENTION In recent decades, the average life expectancy of people has increased significantly, in addition to which individuals have become increasingly interested in beauty and tend to seek to delay or minimize the appearance of signs associated with aging.

[0003] Skin is the largest organ in humans, and because it is located at the interface of the body, it is subject to intrinsic aging (chronologic aging) and extrinsic aging (mainly photoaging). Intrinsic aging is accompanied by cell loss, epidermal thinning, DEJ (dermal-epidermal junction) flattening and fine wrinkles. On the other hand, photoaged skin is wrinkled and pigmented. It is now believed that inflammation and accumulation of ROS (reactive oxygen species) are causative factors in both types of skin aging.

[0004] One of the main functions of the skin is to protect the body from external factors, and it does so by using physical barriers, biomolecules, and a complex network of resident immune and non-immune cells and skin structures. Specifically, the outermost layer of the skin, called the epidermis, provides a barrier that protects against environmental factors, pathogens, heat, UV radiation, and water loss. Keratinocytes are keratin-producing epidermal cells and account for approximately 90% of the epidermal cells (Nguyen AV, Soulika AM (2019). The Dynamics of the Skin's Immune System. International journal of molecular sciences, 20(8), 1811; Lee HS, et al. (2019). Anti-inflammatory effects of ethanol extract from the leaves and shoots of Cedrela odorata L. in cytokine-stimulated keratinocytes. Experimental and therapeutic medicine, 18(1), 833-840).

[0005] Skin ageing is a complex biological process influenced by a combination of intrinsic and exogenous factors (intrinsic factors: genetics, cellular metabolism, hormones and metabolic processes; exogenous factors: chronic light exposure, pollution, ionizing radiation, chemicals and toxins) and thus leads to changes in the skin and the appearance of defects therein (e.g. loss of firmness, wrinkles, roughness and / or sagging), as mentioned above.

[0006] These factors not only induce cumulative structural and physiological changes in the different layers of the skin, but also lead to changes in the overall appearance of the skin (Ganceviciene, R, et al. (2012). Skin Anti-Aging Strategies. Dermato-Endocrinology, 4, 308-319).

[0007] As the aging process occurs, the body's ability to resolve inflammation is significantly reduced, resulting in an imbalance between inflammation and anti-inflammation. "Inflammatory aging" refers to the ongoing low-grade inflammation that accompanies aging. Such chronic inflammatory responses can accumulate over time and gradually cause tissue damage (Zhuang Y and Lyga J (2014) Inflammaging in skin and other tissues - the roles of complement system and macrophage. Inflamm Allergy Drug Targets. 13, 153-61). Inflammatory aging is caused by a systemic increase in multiple inflammatory cytokines and is greatly influenced by several exogenous factors, including ultraviolet radiation (UVR), airborne particulate matter (PM) and the microbiome (Xia S, et al. (2016) An Update on Inflamm-Aging: Mechanisms, Prevention, and Treatment. J Immunol Res. 2016, 8426874).

[0008] In addition, or in parallel, UV radiation, xenobiotics and heat stress disrupt the metabolism of skin cells, resulting in increased production of reactive oxygen species (ROS) and redox imbalance, contributing to oxidative stress resulting in the formation and accumulation of irreversibly damaged proteins, lipids, nucleic acids and carbohydrates in skin cells, which in turn leads to skin aging. The key factors or mechanisms of skin cells against oxidative stress are Nrf2 (which can regulate anti-inflammatory and antioxidant responses through multiple mechanisms, such as regulating redox homeostasis and suppressing inflammatory genes directly or via interaction with NF-κB) and catalase (a key antioxidant enzyme that alleviates oxidative stress to a great extent by destroying cellular hydrogen peroxide to generate water and oxygen).

[0009] Senescence usually occurs in response to damaging stimuli, such as shortening or dysfunction of telomeres (replicative senescence) or persistent activation of the DNA damage response (DDR). The recruitment of an inflammatory phenomenon known as senescence-associated secretory phenomenon (SASP) can promote tissue inflammation and deterioration. Growing evidence suggests that senescent cells accumulate in tissues and organs with aging, thereby impairing physiological regenerative processes and contributing to organismal aging (Rube, CE, et al. (2021). Human skin aging is associated with increased expression of the histone variant H2A.J in the epidermis. Aging Mech Dis 7, 7). Furthermore, as mentioned above, the production of reactive oxygen species increases with aging, causing double-strand breaks (DSBs) in DNA, while skin cells have a decreased ability to repair this damage over time, contributing to chronological aging. During RS (replicative senescence), fibroblasts have been shown to accumulate proteins involved in the DDR of telomeric regions, such as γH2A.X, 53BP1, MDC1 and NBS1 (d'Adda di Fagagna et al., (2003). A DNA damage checkpoint response in telomere-initiated senescence, Nature 426, 194-198). Activation of the DDR can result in the activation of the transcription factor p53, which is involved in various processes including DNA repair and apoptosis, but the most striking consequence is the expression of the cyclin-dependent kinase inhibitor p21, which, together with the activation of p16, is considered to be the main pathway for senescence induction (de Magalhaes JP and Passos JF (2018). Stress, cell senescence and organismal ageing; Mechanisms of Ageing and Development, Volume 170, 2-9). Antioxidants have the ability to bind free radicals caused by oxidative stress, reducing DNA DSBs and, as a result, the appearance of 53BP1 aggregates (foci), preventing and slowing down the aging phenomena of the skin.

[0010] On the other hand, interleukin-8 (IL-8) is a potent chemotactic and inflammatory cytokine that is produced in the skin by various cells in response to inflammatory stimuli. Recent studies suggest that IL-8, in addition to its potent activity on leukocytes, has direct effects on several functions of human epidermal cells, such as chemotaxis, Candida albicans bactericidal activity, or proliferation (Kemeny L et al. (1994). Role of interleukin-8 receptor in skin. International archives of allergy and immunology, 104(4), 317-322).

[0011] Moreover, one of the main players in the innate immune response is the toll-like receptor 2 (TLR2). This protein is a membrane receptor involved in the recognition of pathogen-associated molecular patterns (PAMPs) or exogenous ligands, and also recognizes danger-associated molecular patterns (DAMPs), also known as alarmins, or endogenous ligands. This recognition activates an inflammatory response to protect the organism from external insults (Chen L and DiPietro LA (2017). Toll-Like Receptor Function in Acute Wounds. Adv Wound Care (New Rochelle) 6, 344-355). Focusing on the skin, whose main function is to protect the body from external factors, TLR2 is expressed in many cell types that compose this organ, including keratinocytes (Nguyen AV and Soulika AM (2019). The Dynamics of the Skin's Immune System. International journal of molecular sciences, 20(8), 1811; Pivarcsi A et al. (2003). Expression and function of Toll-like receptors 2 and 4 in human keratinocytes. Int Immunol. 15, 721-30).Dysfunction or excessive activation of TLR2 in the skin may cause excessive inflammatory responses, leading to inflammation-related diseases such as acne, and contribute to the persistent maintenance of an inflammatory state, leading to inflammaging (Bailey KL et al. (2019). Aging leads to dysfunctional innate immune responses to TLR2 and TLR4 agonists. Aging Clin Exp Res. 31, 1185-1193.; Zhang B, et al. (2019) Toll-like receptor 2 plays a critical role in pathogenesis of acne vulgaris. Biomed Dermatol 3, 4).

[0012] Moreover, it is important to highlight that telomeres form the ends of human chromosomes. This structure consists of a simple tandem DNA repeat (TTAGGG), does not code for any gene product, and its main function is to cap the chromosome ends. Telomere capping is necessary to distinguish between chromosome ends that should not be repaired and DNA breaks within the genome, leading to cell cycle arrest and DNA repair or, if the damage is severe or irreparable, the induction of apoptosis. Telomeres prevent the induction of such reactions at the chromosome ends, and this capping is essential for maintaining chromosomal stability, and a minimum telomere length is required to perform this capping function (Buckingham EM and Klingelhutz AJ (2011). The role of telomeres in the ageing of human skin. Exp Dermatol. 20, 297-302; Jiang H et al. (2007). Telomere shortening and ageing. Z Gerontol Geriatr. 40, 314-24).

[0013] Telomeres are shortened with every cell division cycle due to the "end-replication problem" of DNA polymerase, which is one of the most relevant features of aging. Additional factors may be involved in telomere shortening (telomere processing during the cell cycle, reactive oxygen species), and various cellular mechanisms are involved in telomere preservation. Among these mechanisms, the sheltering complex is essential for telomere function, telomere maintenance, and linkage with intracellular signaling pathways. The sheltering complex is a multi-protein structure that protects the telomere ends and is formed by various proteins such as TRF1, TRF2, POT1, RAP1, TIN2, and TPP1 (Imbert I et al. (2012). Modulation of telomere binding proteins: a future area of ​​research for skin protection and anti-aging target. J Cosmet Dermatol. 11, 162-6).

[0014] When telomeres are critically shortened, they lose their capping function at the ends of chromosomes. These dysfunctional telomeres are recognized as DNA damage and induce the DNA damage checkpoint, leading to senescence or even cell death (Imbert I et al. (2012). Modulation of telomere binding proteins: a future area of ​​research for skin protection and anti-aging target. J Cosmet Dermatol. 11, 162-6; Maleki M et al. (2020) Stabilization of telomere by the antioxidant property of polyphenols: Anti-aging potential. Life Sci. 259, 118341). Focusing on skin aging, the accumulation of senescent cells with short telomeres in skin tissue is a hallmark of the aging phenomenon (Buckingham EM and Klingelhutz AJ (2011). The role of telomeres in the ageing of human skin. Exp Dermatol. 20, 297-302; Jiang H et al. (2007). Telomere shortening and ageing. Z Gerontol Geriatr. 40, 314-24).

[0015] Telomerase is an enzyme that extends telomeres, and in cells where it is normally expressed, such as pluripotent stem cells and the adult stem cell compartment, it compensates for telomere loss by adding new TTAGGG repeats to the ends of chromosomes (Blackbum EH (1991). Structure and function of telomeres. Nature 350, 569-573; Liu L, et al. (2007). Telomere lengthening early in development. Nat. Cell Biol. 9:1436- 1441; Flores l et al. (2005). Effects of telomerase and telomere length on epidermal stem cell behavior. Science. 309: 1253-1256; Marion RM et al. (2009). Telomeres acquire embryonic stem cell characteristics in induced pluripotent stem cells. Cell Stem Cell 4:141-54). Although telomerase is expressed in the adult stem cell compartment, this expression is insufficient to counteract the telomere attenuation that accompanies cell division throughout life, and as discussed above, telomeres shorten with age in vitro and in vivo. This progressive shortening of telomeres may impair the regenerative capacity of tissues and has been proposed as one of the molecular hallmarks of aging (Lopez-Otin C et al. (2013). The hallmarks of aging. Cell.153:1194-1217; Martinez P, Blasco MA (2017). Telomere-driven diseases and telomere-targeting therapies. J Cell Biol. 216(4):875-887).Telomere attenuation in stem cells impairs the tissue and self-renewal capabilities of stem cells and is considered to be one of the major molecular causes of aging and the onset of age-related diseases (Flores l et al. (2005). Effects of telomerase and telomere length on epidermal stem cell behavior. Science 309: 1253-1256; Sharpless NE and DePinho RA (2007). How stem cells age and why this makes us grow old. Nat. Rev. Mol. Cell Biol. 8:703-713). In particular, it has been suggested that telomerase plays an important role in maintaining the function and proliferation of skin cells in skin aging (Buckingham EM, Klingelhutz AJ (2011) The role of telomeres in the ageing of human skin. Exp Dermatol. 20, 297-302).

[0016] In recent years, the number of active ingredients for improving the signs of skin aging, such as loss of firmness, skin texture and the appearance of wrinkles, has increased considerably. Examples of such active ingredients are retinoids, vitamins or plant extracts (Bradley EJ et al. (2015). Over-the-counter anti-ageing topical agents and their ability to protect and repair photoaged skin. Maturitas, 80, 265-272).

[0017] Antioxidants are used to reduce the concentration of free radicals in the skin and thus counteract the degradation of collagen. An example of said antioxidant is ascorbic acid (also known as vitamin C). In addition to its antioxidant activity, ascorbic acid promotes epidermal differentiation and induces the synthesis of proteins from the ECM (collagen I and III, and elastin), especially while inhibiting matrix metalloproteinase-1 (MMP1). Unfortunately, ascorbic acid is highly unstable and is oxidized, especially at high temperature, in aerobic conditions, at high pH, ​​and / or when exposed to light (Manela-Azulay M et al. (2017). Vitamins and other Antioxidants. Daily Routine in Cosmetic Dermatology, 1-13).

[0018] Also, a wide range of plant extracts and plant-derived compounds, such as grape extracts containing resveratrol (an antioxidant), green tea containing polyphenols, or soybeans containing isoflavones, have been found in the marketplace with diverse applications. Another example is turmeric (Curcuma longa), known for its use in Chinese and Ayurvedic medicine (as a treatment for inflammatory conditions) and in culinary and beauty rituals around the world. Turmeric contains turmerin, which has been shown to have antioxidant and anti-inflammatory activity.

[0019] However, the in vivo effectiveness and composition of these ingredients have not been fully scientifically verified.

[0020] On the other hand, peptides can also be incorporated into cosmetic formulations to improve the signs of skin aging. Bioactive peptides mimic the body's own molecules and can affect processes such as collagen synthesis, with the advantage of being much better tolerated and stable. Furthermore, bioactive peptides can be deployed in a wide range of activities, chemistries and efficacies (Zhang L and Falla TJ (2009). Cosmeceuticals and peptides. Clinics in dermatology, 27, 485-494).

[0021] Despite the wide variety of compounds and / or extracts present in the field, there remains a need to find new molecules, preferably peptides, that are able to provide prevention and / or treatment of ageing, preferably skin ageing, and that exhibit a broad spectrum of activity against said ageing and diseases associated with ageing, inflammation and oxidative stress.

[0022] Description of the Invention BRIEF DESCRIPTION OF THE PRESENT EMBODIMENT After extensive and intensive research, the inventors of the present invention have surprisingly found peptides with a broad spectrum of activity, including telomere protection as well as antioxidant and anti-inflammatory activity, etc. Therefore, these peptides of the present invention are useful in the treatment of skin aging and symptoms related to said skin aging, as well as inflammatory diseases and diseases related to oxidative stress, thus solving the problems existing in the prior art mentioned above.

[0023] BRIEF DESCRIPTION OF THE DRAWINGS For a better understanding, the invention will now be described in more detail with reference to the accompanying drawings, given by way of example, and with reference to illustrative and non-limiting examples, in which: * means p<0.05; ** means p<0.01, *** means p<0.001.

[0024] Figure 1 shows the gene expression results obtained in Example 2 when HEKa cells were treated with 0.01 mg / mL of peptide of SEQ ID NO:2-NH2 for 6 hours. Each bar represents one gene from top to bottom: TOLLIP, IRAK3, TNFAIP3 and HMOX1. Bars extending to the right (i.e., positive values) indicate upregulation of the gene.

[0025] Figure 2 shows the gene expression results obtained in Example 2 when HEKa cells were treated with 0.01 mg / mL of peptide SEQ ID NO:2-NH2 for 24 hours. Each bar represents one gene from top to bottom: NQO1, NFKB1, IL1B, TNF, CXCL8, IL6, MMP3 and TLR2. Bars extending to the left (i.e., negative values) indicate downregulation of the gene. Bars extending to the right (i.e., positive values) indicate upregulation of the gene.

[0026] Figure 3 shows the miRNA expression results obtained in Example 3 when HEKa cells were treated with 0.01 mg / mL of peptide of SEQ ID NO: 2-NH2 for 24 hours. Each bar represents one miRNA from top to bottom: miR-146a-5p and miR-21-5p. Bars extending to the right (i.e., positive values) indicate upregulation of genes.

[0027] Figure 4 shows the results obtained in Example 4 for the secretion or release of IL-8 under inflammatory conditions (except for untreated or basal samples) when HaCaT keratinocytes are treated with the peptide of SEQ ID NO: 2-NH2. The results in this figure are shown relative to the positive control sample (cells treated with 10 ng / mL of IFNγ), in which the secretion of IL-8 is taken as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: negative control (basal), positive control, cells treated with 10 ng / mL of IFNγ and 0.005 mg / mL, 0.01 mg / mL or 0.05 mg / mL of the peptide of SEQ ID NO: 2-NH2. The y-axis represents the percentage (%) of secreted IL-8 relative to the positive control (set as 100%).

[0028] Figure 5 shows the results obtained in Example 4 for the secretion or release of IL-8 under inflammatory conditions (except for untreated or basal samples) when HaCaT keratinocytes are treated with the peptide of SEQ ID NO: 3-NH2. The results in this figure are shown relative to the positive control sample (cells treated with 10 ng / mL of IFNγ), in which the secretion of IL-8 is taken as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: negative control (basal), positive control, cells treated with 10 ng / mL of IFNγ and 0.005 mg / mL, 0.01 mg / mL or 0.05 mg / mL of the peptide of SEQ ID NO: 3-NH2. The y-axis represents the percentage (%) of secreted IL-8 relative to the positive control (set as 100%).

[0029] Figure 6 shows the results obtained in Example 4 for the secretion or release of IL-8 under non-inflammatory conditions when HaCaT keratinocytes are treated with the peptide of SEQ ID NO: 2-NH2. The results in this figure are shown relative to the basal state (untreated cells), in which the secretion of IL-8 is taken as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: negative control (basal), cells treated with 0.005 mg / mL, 0.01 mg / mL or 0.05 mg / mL of the peptide of SEQ ID NO: 2-NH2. The y-axis represents the percentage (%) of secreted IL-8 relative to the basal state (set as 100%).

[0030] Figure 7 shows the results obtained in Example 4 for the secretion or release of IL-8 under non-inflammatory conditions when HaCaT keratinocytes are treated with the peptide of SEQ ID NO: 3-NH2. The results in this figure are shown relative to the basal state (untreated cells), in which the secretion of IL-8 is taken as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: negative control (basal), cells treated with 0.005 mg / mL, 0.01 mg / mL or 0.05 mg / mL of the peptide of SEQ ID NO: 3-NH2. The y-axis represents the percentage (%) of secreted IL-8 relative to the basal state (set as 100%).

[0031] FIG. 8 shows the results obtained for telomere length when HDFa cells were treated with the peptide of SEQ ID NO: 2-NH2 in Example 5. The results are shown for the telomere length of 25-year-old subjects (HDFa 25 The telomere lengths are expressed as a percentage (%) of the telomere length of HDFa in untreated control mice. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: untreated HDFa 25 Cells, untreated HDFa 73 Cells, HDFa treated with 0.005mg / mL or 0.01mg / mL, respectively 73 Cells (HDFa from a 73-year-old subject). The y-axis shows untreated HDFa 25 The percentage (%) of telomere length relative to the cells (set as 100%) is shown.

[0032] Figure 9 shows the results obtained in Example 6 for Nrf2 translocation into the nucleus when HEKa cells are treated with the peptide of SEQ ID NO: 2-NH2. The results are expressed as a percentage (%) of the amount of nucleolar Nrf2 in comparison with the amount of nucleolar Nrf2 in untreated cells (negative control - basal state), which is set as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: untreated cells and cells treated with 0.01 and 0.05 mg / mL of the peptide of the invention. The y-axis represents the amount of nucleolar Nrf2 in percentage (%) relative to untreated cells, which is set as 100%.

[0033] Figure 10 shows the results obtained for TLR2 abundance when HEKa cells are treated with peptide of SEQ ID NO: 2-NH2 in Example 7. Results are expressed as percentage (%) of TLR2 abundance compared to untreated cells (negative control - basal state), which is set as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: untreated cells; cells treated with 10 ng / mL IFNγ; and cells treated with 10 ng / mL IFNγ and 0.01 or 0.05 mg / mL of the peptide of the invention, respectively. The y-axis represents the TLR2 abundance as percentage (%) relative to the TLR2 abundance of untreated cells, which is set as 100%.

[0034] Figure 11 shows the results obtained for the amount or synthesis of catalase when HEKa cells are treated with the peptide of SEQ ID NO: 2-NH2 in Example 8. The results are expressed as the amount of catalase relative to the total protein amount of the sample and as a percentage (%) relative to the value obtained for the basal condition (untreated cells), which is set as 100%. The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: untreated cells (basal condition); and cells treated with 0.005 mg / mL, 0.01 mg / mL, or 0.05 mg / mL of the above peptide. The y-axis represents the percentage (%) of the amount of catalase / total protein relative to the basal condition, which is set as 100%.

[0035] Figure 12 shows the results obtained in Example 9 for TRF1 production (TRF1 amount measured by the measured intensity) when CHA9.3 cells are treated with peptides of SEQ ID NO:2-NH2 and SEQ ID NO:3-NH2. The results are normalized with respect to the negative control (cells treated with vehicle, i.e. DMSO only). The columns or bars on the x-axis show, from left to right, the different experimental treatment groups: negative control (cells treated with DMSO), cells treated with peptide of SEQ ID NO:3-NH2 at 0.063 mg / mL, cells treated with peptide of SEQ ID NO:2-NH2 at 0.125 mg / mL. The y-axis shows the TRF1 intensity obtained relative to the negative control (TRF1 intensity is set as 100% for the negative control group, and the values ​​of the remaining treatment groups are set by comparison with the negative control mentioned above).

[0036] Figure 13 shows the results obtained for TRF1 aggregates (foci) per cell when HEKa cells are treated with the peptide of SEQ ID NO:2-NH2 in Example 10. The results are normalized with respect to the negative control (basal state, cells not treated with the peptide of the invention). The columns or bars on the x-axis show, from left to right, the different experimental treatment groups: negative control (basal state) and cells treated with 0.005 mg / mL of the peptide of SEQ ID NO:2-NH2. The y-axis shows the TRF1 aggregates per cell relative to the negative control (the number of TRF1 aggregates per cell is set as 100% for the negative control group, and the values ​​for the remaining treatment groups are set by comparison with the negative control above).

[0037] Figure 14 shows the results obtained for DNA protection (measured based on 53BP1 aggregates per cell) obtained in Example 10, where HEKa cells were treated with the peptide of SEQ ID NO: 2-NH2. The results were normalized with respect to the positive control (cells treated only with H2O2). The columns or bars on the x-axis represent, from left to right, the different experimental treatment groups: negative control (basal condition, untreated cells), positive control (cells treated only with H2O2), cells treated with H2O2 and 0.01 mg / mL or 0.05 mg / mL of the peptide of the invention. The y-axis represents the 53BP1 aggregates per cell relative to the positive control (the number of 53BP1 aggregates per cell is set as 100% for the positive control group, and the values ​​for the remaining treatment groups are set by comparison with the negative control mentioned above).

[0038] Detailed Description of the Invention In a first aspect, the present invention relates to peptides with a broad spectrum of activity against skin ageing, including telomere protection as well as anti-inflammatory and antioxidant activity.

[0039] In a second aspect, the present invention relates to a composition comprising a peptide of the invention.

[0040] In a further aspect, the present invention relates to the cosmetic use of a peptide of the invention or a composition of the invention in a subject in need of treatment.

[0041] In a fourth aspect, the present invention relates to the cosmetic use of a peptide of the invention or a composition of the invention in a subject in need of treatment.

[0042] In a fifth aspect, the present invention relates to a method for the prevention and / or treatment of skin ageing in a subject, comprising administering to the subject a peptide of the invention or a composition of the invention.

[0043] In a further aspect, the present invention relates to a composition or a peptide of the invention for use as a medicament.

[0044] In a seventh aspect, the present invention relates to the use of a composition or peptide of the invention for the manufacture of a medicament for the prevention, amelioration and / or treatment of an age-related disease, an inflammatory disease or a disease associated with oxidative stress.

[0045] In an eighth aspect, the present invention relates to a method for the prevention, amelioration and / or treatment of an age-related disease, an inflammatory disease or a disease associated with oxidative stress, comprising administering to a subject in need thereof a peptide or composition of the present invention.

[0046] In a ninth aspect, the present invention relates to a dietary supplement composition comprising a peptide of the present invention.

[0047] In a final aspect, the present invention relates to the use of a peptide of the invention or a composition of the invention as a dietary supplement.

[0048] As used herein, the term "acyclic aliphatic group" and its plurals have the ordinary meaning given to such terms in the art. Thus, these terms refer to, for example, but not limited to, linear or branched alkyl, alkenyl, and alkynyl groups.

[0049] As used herein, the term "alkyl group" and its plural forms refer to a saturated, straight or branched group having 1 to 24, preferably 1 to 16, more preferably 1 to 14, even more preferably 1 to 12, and even more preferably 1, 2, 3, 4, 5, or 6 carbon atoms and attached to the remainder of the molecule by a single bond, such as, but not limited 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, etc. Alkyl groups may be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, alkoxythio, etc.

[0050] The term "alkenyl group" and its plural forms as used herein refers to a linear or branched group having 2 to 24, preferably 2 to 16, more preferably 2 to 14, even more preferably 2 to 12, and even more preferably 2, 3, 4, 5, or 6 carbon atoms, one or more carbon-carbon double bonds, preferably 1, 2, or 3 carbon-carbon double bonds, conjugated or unconjugated, and bonded to the remainder of the molecule through a single bond, such as, but not limited to, vinyl, oleyl, linoleyl, and the like. The alkenyl group may be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0051] As used herein, the term "alkynyl group" and its plural forms refer to a linear or branched group having 2 to 24, preferably 2 to 16, more preferably 2 to 14, even more preferably 2 to 12, and even more preferably 2, 3, 4, 5, or 6 carbon atoms, one or more carbon-carbon triple bonds, preferably 1, 2, or 3 carbon-carbon triple bonds that are conjugated or unconjugated, and bonded to the remainder of the molecule through a single bond, such as, but not limited to, ethynyl groups, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, and the like. The alkynyl group may be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0052] As used herein, the term "alicyclic group" and its plural forms have the ordinary meaning given to such terms in the art. Thus, these terms are used to refer to, for example, but not limited to, cycloalkyl, cycloalkenyl, or cycloalkynyl groups.

[0053] As used herein, the term "cycloalkyl" and its plural forms refer to a saturated monocyclic or polycyclic aliphatic group having 3 to 24, preferably 3 to 16, more preferably 3 to 14, even more preferably 3 to 12, and even more preferably 3, 4, 5, or 6 carbon atoms, attached to the remainder of the molecule via a single bond, such as, but not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, methylcyclohexyl, dimethylcyclohexyl, octahydroindene, decahydronaphthalene, dodecahydrophenalene, adamantyl, and the like. Cycloalkyls can be optionally substituted with one or more groups, such as alkyl, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0054] The term "cycloalkenyl" and its plural forms as used herein refers to a non-aromatic monocyclic or polycyclic aliphatic group having 5 to 24, preferably 5 to 16, more preferably 5 to 14, even more preferably 5 to 12, and even more preferably 5 or 6 carbon atoms, one or more carbon-carbon double bonds, preferably 1, 2 or 3 carbon-carbon double bonds that are conjugated or non-conjugated, and bonded to the remainder of the molecule via a single bond, such as, but not limited to, cyclopent-1-en-1-yl and similar groups. Cycloalkenyl can be optionally substituted with one or more groups, such as alkyl, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0055] The term "cycloalkynyl" and its plural forms as used herein refers to a non-aromatic monocyclic or polycyclic aliphatic group having 8 to 24, preferably 8 to 16, more preferably 8 to 14, even more preferably 8 to 12, and even more preferably 8 or 9 carbon atoms, one or more carbon-carbon triple bonds, preferably 1, 2 or 3 carbon-carbon triple bonds that are conjugated or non-conjugated, and bonded to the remainder of the molecule via a single bond, such as, but not limited to, cyclooct-2-yn-1-yl groups. Cycloalkynyl can be optionally substituted with one or more groups, such as alkyl, halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0056] As used herein, the term "aryl group" and its plural forms refer to an aromatic group having 6 to 30, preferably 6 to 18, more preferably 6 to 10, and even more preferably 6 or 10 carbon atoms, having 1, 2, 3 or 4 aromatic rings, bonded or fused with carbon-carbon bonds, and bonded to the remainder of the molecule through a single bond, including, for example, but not limited to, phenyl, naphthyl, diphenyl, indenyl, phenanthryl, or anthranyl, which may be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0057] As used herein, the term "aralkyl group" and its plurals refer to an alkyl group having 7 to 24 carbon atoms and substituted with an aromatic group, including, but not limited to, -(CH2)1-6-phenyl, -(CH2)1-6-(1-naphthyl), -(CH2)1-6-(2-naphthyl), -(CH2)1-6-CH(phenyl)2, and the like. Aralkyl groups can be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0058] As used herein, the term "heterocyclic group" and its plural forms refer to a 3- to 10-membered heterocyclyl or hydrocarbon ring in which one or more of the ring atoms, preferably one, two or three of the ring atoms, are elements different from carbon, such as nitrogen, oxygen or sulfur, and may be saturated or unsaturated. For the purposes of this invention, a heterocyclyl may be a cyclic, monocyclic, bicyclic or tricyclic system, which may include fused ring systems; and the nitrogen, carbon or sulfur atoms may be optionally oxidized in the heterocyclyl radical; the nitrogen atom may be optionally quaternized; and the heterocyclyl radical may be partially or fully saturated or aromatic. Preferably, the term heterocyclyl relates to a 5- or 6-membered ring. The heterocyclic group may be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto and alkoxythio.

[0059] As used herein, the term "heteroarylalkyl group" and its plurals refer to an alkyl group substituted with a substituted or unsubstituted aromatic heterocyclyl group, the alkyl group having 1 to 6 carbon atoms, the aromatic heterocyclyl group having 2 to 24 carbon atoms and 1 to 3 non-carbon atoms, such as, but not limited to, -(CH2)1-6-imidazolyl, -(CH2)1-6-triazolyl, -(CH2)1-6-thienyl, -(CH2)1-6-furyl, -(CH2)1-6-pyrrolidinyl, and the like. Heteroarylalkyl groups can be optionally substituted with one or more substituents, such as halo, hydroxy, alkoxy, carboxy, carbonyl, cyano, acyl, alkoxy-carbonyl, amino, nitro, mercapto, and alkoxythio.

[0060] As used herein, the terms "halo" or "halogen" refer to fluorine, chlorine, bromine, or iodine, the anions of which are referred to as halides.

[0061] The term "derivatives" and its plural forms as used herein refer to cosmetically acceptable, pharma- ceutically acceptable and / or dietarily acceptable compounds, i.e. compounds derived from the compound of interest that can be used in the preparation of cosmetic, pharmaceutical and / or dietary compositions, as well as to cosmetically unacceptable, pharma-ceutically unacceptable and / or dietarily unacceptable derivatives, since the latter may be useful in the preparation of cosmetically acceptable, pharma-ceutically acceptable and / or dietarily acceptable derivatives.

[0062] The term "salt" and its plural forms as used herein refer to any type of salt known in the art, such as halide salts, hydroxy acid salts (oxy acid salts, acid salts, base salts and double salts, etc.), hydroxy salts, mixed salts, oxy salts or other hydrated salts. The term includes cosmetically acceptable, pharma- ceutically acceptable and / or food acceptable salts; as well as cosmetically unacceptable, pharma- ceutically unacceptable and / or food unacceptable salts, since the latter may be useful in the preparation of cosmetically acceptable, pharma- ceutically acceptable and / or food acceptable salts.

[0063] As used herein, the term "isomer" and its plural forms refer to optical isomers, enantiomers, stereoisomers or diastereoisomers. Individual enantiomers or diastereoisomers, as well as mixtures thereof, can be separated by conventional techniques known in the art.

[0064] The term "solvate" and its plural forms as used herein refer to any solvate known in the art, such as polar, non-polar or amphiphilic solvates, including any cosmetically acceptable, pharma- ceutical and / or food acceptable solvates that provide the compound of interest (peptide of the present invention) when administered or applied (directly or indirectly) to a subject of interest.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), more preferably a hydrate, or a solvate with an alcohol such as ethanol.

[0065] Furthermore, as used herein, the term "amino acid" and its plural forms include not only the amino acids encoded by the genetic code, but also the non-encoded amino acids, whether they are naturally occurring and whether they are D-amino acids and L-amino acids. Examples of non-encoded amino acids include, but are not limited to, 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, hydroxyproline, alloisoleucine, alloiisothreonine, isonipecotic acid, isoserine, phenylglycine, statins, β-alanine, norleucine, N-methyl amino acids, α-amino acids and β-amino acids, and derivatives thereof, among others. Nonetheless, additional unnatural amino acids are known in the art (see, e.g., "Unusual amino acids in peptide synthesis" by Roberts DC and Vellaccio F (1983) The Peptides, Vol. 5, Chapter VI. Gross E and Meienhofer J, Eds., Academic Press, New York, USA). Amino acid abbreviations used herein are in accordance with the 1983 IUPAC-IUB Joint Commission on Biochemical Nomenclature recommendations outlined in Eur. J. Biochem. (1984) 138:937.

[0066] "Percentage of identity" as used herein with respect to peptides, polypeptides and proteins has the meaning generally ascribed to them in the art, and thus refers to the percentage of amino acids that are identical between two amino acid sequences compared after optimal alignment of these sequences, where the percentage is merely statistical and the differences between the two amino acid sequences are randomly distributed throughout the sequences. "Optimal alignment" is understood as the alignment of amino acid sequences that results in a larger percentage of identity. The percentage of identity is calculated by determining the number of identical positions in the two compared sequences where the amino acids are identical, dividing the number of identical positions by the number of positions compared, and multiplying the result by 100 to obtain the percentage of identity between the two sequences. The sequence comparison between two amino acid sequences can be performed manually or using computer programs known in the art, such as the BLAST (Basic Local Alignment Search Tool) algorithm.

[0067] The terms "inflammaging" and "inflamm-aging" are used interchangeably herein and have the meanings they commonly have in the art, i.e., referring to the chronic, low-grade inflammation that characterizes aging.

[0068] Thus, in a first aspect, the present invention provides a peptide of formula (I): R1-AA1-AA2-AA3-AA4-AA5-AA6-AA7-AA8-R2 (I) (R1-SEQ ID NO:1-R2) and its acceptable isomers, salts, solvates, derivatives and / or mixtures thereof, AA1 is a small amino acid; AA2 is selected from a polar uncharged amino acid or an aromatic amino acid; AA3 is absent or selected from aromatic amino acids; AA4 is selected from negatively charged amino acids; AA5 is selected from nonpolar amino acids; AA6 is selected from Asn or Leu; AA7 is selected from negatively charged amino acids; AA8 is absent or selected from aromatic amino acids; R1 is selected from H, substituted or unsubstituted acyclic aliphatic, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and R5-CO-, where R5 is a substituted or unsubstituted C1-C 24 Alkyl radicals, substituted or unsubstituted C2-C 24 Alkenyl, substituted or unsubstituted C2-C 24 Alkynyl, substituted or unsubstituted C3-C 24 Cycloalkyl, substituted or unsubstituted C5-C 24 Cycloalkenyl, substituted or unsubstituted C8-C 24 Cycloalkynyl, substituted or unsubstituted C6-C 30 Aryl, substituted or unsubstituted C7-C 24 aralkyl, a substituted or unsubstituted 3- to 10-membered heterocyclyl ring, and a substituted or unsubstituted heteroarylalkyl having 2 to 24 carbon atoms, 1 to 3 non-carbon atoms, and an alkyl chain of 1 to 6 carbon atoms; and R2 is selected from H, -NR3R4-, -OR3 and -SR3, where R3 and R4 are independently selected from H, a substituted or unsubstituted acyclic aliphatic group, a substituted or unsubstituted alicyclyl, a substituted or unsubstituted heterocyclyl, a substituted or unsubstituted heteroarylalkyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted aralkyl; Alternatively, R1 and R2 are absent and AA1 refers to the peptide, directly linked to the last amino acid of the peptide, ie, AA8, or AA7 if AA8 is absent (forming a cyclic peptide).

[0069] R1 is linked to the N-terminus of the peptide, ie, the amino group of AA1 (R1 replaces or exchanges an H from the amino group of AA1).

[0070] R2 is linked to the C-terminus of the peptide, i.e., to the carboxy group of AA8, or, if AA8 is absent, to the carboxy group of AA7 (R2 replaces or replaces the OH of the carboxy group).

[0071] 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.

[0072] Preferably, the above-mentioned isomer is a stereoisomer. Said stereoisomer is intended to be an enantiomer or a diastereoisomer. Thus, in a preferred embodiment of the present invention, the peptide is a racemic mixture, a diastereomeric mixture, a pure enantiomer or a pure diastereoisomer.

[0073] Furthermore, the isomers, salts, solvates, derivatives and / or mixtures thereof are preferably cosmetically acceptable, pharma- ceutically acceptable and / or food acceptable isomers, salts, solvates, derivatives and / or mixtures thereof.

[0074] Preferably, AA1 is selected from Cys, Val, Ala, Pro, Ser, Asn, Ala, Gly or Thr, more preferably, AA1 is selected from Ser, Cys, Ala or Gly; even more preferably, AA1 is Gly.

[0075] Also preferably, AA2 is selected from Tyr, Phe, Trp, Ser, Thr, Gln, Asn, Cys or Gly, more preferably, AA2 is selected from Tyr, Trp, Ser, Asn or Gln, more preferably, AA2 is selected from Tyr or Asn.

[0076] Preferably, AA3 is absent or selected from Tyr, Phe or Trp, more preferably, AA3 is absent or Tyr.

[0077] In the present invention, AA4 is preferably selected from Asp or Glu, and more preferably, AA4 is Glu.

[0078] Also preferably, AA5 is selected from Ala, Val, Ile, Leu, Met, Pro, Phe or Trp, more preferably, AA5 is selected from Ala, Ile, Leu or Val, and even more preferably, AA5 is Leu.

[0079] In the present invention, AA7 is preferably selected from Asp or Glu, and more preferably, AA7 is Asp.

[0080] Preferably, AA8 is absent or selected from Tyr, Phe or Trp, more preferably, AA8 is absent or Tyr.

[0081] Thus, in a preferred embodiment, in formula (I): AA1 is selected from Cys, Val, Ala, Pro, Ser, Asn, Ala, Gly, or Thr; AA2 is selected from Tyr, Ser, Thr, Gln, Asn, Cys, or Gly; AA3 is absent or selected from Tyr, Phe, or Trp; AA4 is selected from Asp or Glu; AA5 is selected from Ala, Val, Ile, Leu, Met, Pro, Phe, or Trp; AA6 is selected from Asn or Leu; AA7 is selected from Asp or Glu; AA8 is absent or selected from Tyr, Phe, or Trp; More preferably, in formula (I): AA1 is selected from Cys, Val, Ala, Ser, Asn, Ala, Gly, or Thr; AA2 is selected from Tyr, Ser, Thr, Gln, Asn, Cys, or Gly; AA3 is absent or selected from Tyr, Phe, or Trp; AA4 is selected from Asp or Glu; AA5 is selected from Val, Ile, Leu, Met, Pro, Phe, or Trp; AA6 is selected from Asn or Leu; AA7 is selected from Asp or Glu; AA8 is absent or selected from Tyr, Phe, or Trp; More preferably, in formula (I): AA1 is selected from Ser, Cys, Ala, or Gly; AA2 is selected from Tyr, Trp, Ser, Gln or Asn (more preferably, Tyr or Asn); AA3 is absent or selected from Tyr, Phe or Trp (more preferably absent or Tyr); AA4 is selected from Asp or Glu (more preferably, Glu); AA5 is selected from Ala, Val, Ile, or Leu (more preferably, Leu); AA6 is selected from Asn or Leu; AA7 is selected from Asp or Glu (more preferably, Asp); AA8 is absent or selected from Tyr, Phe or Trp (more preferably absent or Tyr); Even more preferably, in formula (I): AA1 is Gly; AA2 is selected from Tyr or Asn; AA3 is absent or Tyr; AA4 is Glu; AA5 is Leu; AA6 is selected from Asn or Leu; AA7 is Asp; AA8 is absent or Tyr.

[0082] In any of the above embodiments, if AA3 is absent, then AA8 is preferably present, and vice versa, i.e., in a preferred embodiment, only one of AA3 and AA8 is absent.

[0083] Thus, in one embodiment, AA3 and AA8 are both present. In another embodiment, AA3 and AA8 are both absent. However, in a most preferred embodiment, AA3 is absent and AA8 is present; and in another most preferred embodiment, AA3 is present and AA8 is absent.

[0084] In a preferred embodiment, the peptide of the invention is: R1-Gly-Tyr-Tyr-Glu-Leu-Asn-Asp-R2 (R1-SEQ ID NO:2-R2); or R1-Gly-Asn-Glu-Leu-Leu-Asp-Tyr-R2 (R1-SEQ ID NO:3-R2), R1 and R2 are as disclosed above.

[0085] More preferably, the peptide of the invention is: R1-L-Gly-L-Tyr-L-Tyr-L-Glu-L-Leu-L-Asn-L-Asp-R2 (R1-SEQ ID NO:2-R2); or R1-L-Gly-L-Asn-L-Glu-L-Leu-L-Leu-L-Asp-L-Tyr-R2 (R1-sequence number 3-R2).

[0086] In any of the above embodiments, preferably, R1 is H or acetyl, and more preferably, R1 is H.

[0087] Also, in any of the above embodiments, R2 is NH2 or OH, more preferably R2 is NH2.

[0088] Thus, in the most preferred embodiment, R1 is H and R2 is NH2.

[0089] In a most preferred embodiment, the peptide of the invention is: Gly-Tyr-Tyr-Glu-Leu-Asn-Asp-NH2 (SEQ ID NO:2-NH2); or Gly-Asn-Glu-Leu-Leu-Asp-Tyr-NH2 (sequence number 3-NH2).

[0090] More preferably, the peptide of the invention has the structure: L-Gly-L-Tyr-L-Tyr-L-Glu-L-Leu-L-Asn-L-Asp-NH2 (SEQ ID NO:2-NH2); or L-Gly-L-Asn-L-Glu-L-Leu-L-Leu-L-Asp-L-Tyr-NH2 (sequence number 3-NH2).

[0091] The peptide of SEQ ID NO: 2 is a biomimetic peptide derived or generated from turmeric protein (Curcuma longa) - UniProt ref P85278 (TURM_CURLO).

[0092] The peptide of SEQ ID NO: 3 is a biomimetic peptide derived or generated from the ricin protein (Ricinus communis (castor bean)) - UniProt ref P02879 (RICI_RICCO).

[0093] In a preferred embodiment, which may be combined with any of the above embodiments, the peptide of the invention provides telomere protection, preferably by increasing the expression and / or amount of TRF1.

[0094] In a further preferred embodiment, which can be combined with any of the above embodiments, the peptide of the invention provides increased or restored length at telomeres.

[0095] In a further preferred embodiment, which can be combined with any of the above embodiments, the peptide of the present invention provides antioxidant activity, preferably by increasing the amount of catalase, by translocation of Nfr2 into the nucleus of the cell, or a combination thereof, more preferably by increasing the amount of catalase and by translocation of Nfr2 into the nucleus of the cell.

[0096] In a further preferred embodiment, which can be combined with any of the above embodiments, the peptide of the present invention provides anti-inflammatory activity, preferably by modulating inflammation-related genes and / or miRNAs, reducing the amount of IL-8 and / or reducing the amount of TLR2, or a combination thereof, most preferably by reducing at least the amount of IL-8.

[0097] The modulation of inflammation-related genes is preferably the upregulation of the genes TNFAIP3, TOLLP, HMOX1, IRAK3, NQO1 and the downregulation of the genes NFKB1, IL1B, TNF, CXCL8, IL6, MMP3 and TLR2.

[0098] The modulation of inflammation-related miRNAs is preferably the upregulation of miR-146a-5p and miR-21-5p.

[0099] Also included within the scope of the present invention are peptides derived or obtained from any of the above disclosed peptides, preferably incorporating one or more conservative substitutions, so long as they retain, exhibit or improve at least one of the aforementioned activities (and preferably all of the activities described above for the peptides of the present invention).

[0100] Additionally, within the scope of the present invention are peptides having a percentage identity of 70%, preferably 80%, more preferably 90%, more preferably 95%, and even more preferably 99% with any of the above-mentioned specific sequences, so long as they retain, exhibit or improve at least one of the above-mentioned activities (and preferably all of the activities described above for the peptides of the present invention).

[0101] As can be directly derived from the above, it is contemplated that the peptides of the present invention are linear or cyclic peptides.

[0102] In one embodiment, which can be combined with any of the above embodiments, the peptide is cyclic. In this case, the first amino acid and the last amino acid (as explained above) of the peptide of the present invention are bonded. Therefore, appropriate modifications to any of the above embodiments must be made (e.g., R1 and R2 may not be present, since the amino acids to which they are bonded are now bonded to each other).

[0103] In a preferred embodiment, which may be combined with any of the above embodiments, the peptide is linear.

[0104] The peptides of the present invention can be synthesized and produced by any means known in the art. For example, they can be synthesized and produced by chemical synthesis (preferably by solid-phase peptide synthesis), expression of said peptides in cell culture, or transgenic production of peptides in plants or animals. Furthermore, the peptides of the present invention can be purified by any means known in the art.

[0105] In a preferred embodiment, which may be combined with any of the embodiments disclosed above, the peptide of the present invention is suitable or adapted to be applied by iontophoresis, more preferably to the face of a subject, preferably a human.

[0106] In a further preferred embodiment, which may be combined with any of the embodiments disclosed above, the peptide of the present invention is suitable or adapted to be applied subcutaneously, more preferably to the face of a subject, preferably a human.

[0107] In another preferred embodiment, which may be combined with any of the embodiments disclosed above, the peptide of the present invention is suitable or adapted to be applied topically (preferably in the form of a cream), more preferably to the face of a subject (preferably a human).

[0108] As can be directly derived from the examples contained below, the peptides of the invention provide a broad spectrum of activity useful in cosmetics for the prevention and / or treatment of skin aging and inflammatory aging: -Protection of telomeres. -Lengthening of telomeres. -Protective against inflammation or anti-inflammatory activity. -Antioxidant activity. -Protection against heat.

[0109] Thus, the peptides of the present invention provide a broad range of activity for the prevention and / or treatment of inflammatory aging and skin aging, solving the above-mentioned problems existing in the art.

[0110] Furthermore, the peptides of the present invention are also useful as medicines due to the above-mentioned activities (especially anti-inflammatory and antioxidant activities), which indicate that the peptides of the present invention are useful for treating, for example, age-related diseases, inflammatory diseases and / or diseases associated with oxidative stress.

[0111] Furthermore, due to the above-mentioned activities (especially the anti-inflammatory and antioxidant activities), which are demonstrated in the examples included below, the peptides of the present invention are also useful in the food industry, preferably as dietary supplements.

[0112] In a second aspect, the present invention relates to a composition comprising a peptide of the invention.

[0113] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0114] The composition of the present invention is preferably a cosmetic composition. Therefore, in this embodiment, the cosmetic composition of the present invention contains at least one peptide of the present invention in a cosmetically effective amount, more preferably, the cosmetic composition of the present invention contains 0.0001% (weight / volume in g / 100 mL, hereinafter, w / v) to 0.05% (w / v) of the peptide of the present invention. In the most preferred embodiment, the cosmetic composition of the present invention contains 0.0001% (w / v) to 0.005% (w / v), more preferably, 0.0005% (w / v) to 0.0015% (w / v) of the peptide of the present invention.

[0115] The cosmetic composition of the present invention is also intended to include 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 art, as long as it does not affect the activity of the peptide of the present invention or does not affect it in an unacceptable manner.

[0116] In another embodiment, the composition of the invention is a pharmaceutical composition.

[0117] The pharmaceutical composition of the present invention is also intended to include at least one additional pharmaceutical ingredient. Said additional pharmaceutical ingredient can be at least one excipient and / or at least one additional active pharmaceutical ingredient. Said additional pharmaceutical ingredient can be any cosmetic ingredient known in the art, as long as it does not affect the activity of the peptide of the present invention or does not affect it in an unacceptable manner.

[0118] In a further embodiment, the composition of the invention is a food composition.

[0119] The food composition of the present invention is also intended to include at least one food ingredient. The additional food ingredient may be any food ingredient known in the art, so long as it does not affect or does not affect in an unacceptable manner the activity of the peptide of the present invention.

[0120] Preferably, the food composition of the present invention is a dietary supplement.

[0121] In one embodiment, a composition of the invention consists essentially of a peptide of the invention.

[0122] In another embodiment, the composition of the invention consists of the peptide of the invention.

[0123] In a third aspect, the present invention relates to the cosmetic use of a peptide of the invention or a composition of the invention in a subject in need of treatment.

[0124] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0125] The composition of the present invention is as disclosed above in the second aspect of the invention.

[0126] The peptides or compositions of the present invention are used in a cosmetically effective amount or quantity.

[0127] The composition of the present invention is preferably a cosmetic composition as disclosed above in the second aspect of the present invention.

[0128] In a preferred embodiment, the cosmetic use is to prevent, treat and / or reduce skin aging, more preferably to reduce, prevent and / or remove the signs of skin aging.

[0129] Preferably, the signs of skin aging are wrinkles, rough skin (preferably rough wrinkles), loose skin or sagging skin, more preferably wrinkles, rough skin, non-inflammatory loose skin or non-inflammatory sagging skin, more preferably wrinkles, even more preferably facial wrinkles. Said wrinkles may be aging wrinkles or expression (muscle origin) wrinkles.

[0130] Non-inflammatory skin laxity corresponds to cosmetic skin laxity, ie, a non-pathological (non-medical) condition.

[0131] Non-inflammatory loose skin corresponds to cosmetic loose skin, ie, a non-pathological (non-medical) condition.

[0132] In another preferred embodiment, the cosmetic use is the prevention, treatment and / or reduction of inflammatory ageing, more preferably the prevention, treatment and / or reduction of inflammatory ageing of the skin.

[0133] In this third aspect of the invention, the subject in need of treatment is preferably a mammal, more preferably a human.

[0134] In this third aspect of the invention, it is also contemplated that the cosmetic use is for improving skin firmness and / or elasticity.

[0135] As mentioned above, in a fourth aspect, the present invention relates to the cosmetic use of a peptide of the invention or a composition of the invention in a subject in need of treatment.

[0136] Preferably, in this fourth aspect of the invention, the use of the cosmetic product is to prevent, treat and / or reduce skin aging, more preferably to reduce, prevent and / or remove the signs of skin aging.

[0137] In another preferred embodiment, the cosmetic use is to prevent, treat and / or reduce inflammatory aging, more preferably cutaneous inflammatory aging, where inflammatory aging is cosmetic inflammatory aging, preferably cosmetic cutaneous inflammatory aging.

[0138] In this fourth aspect of the invention, it is also contemplated that the cosmetic use is for improving skin firmness and / or elasticity.

[0139] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0140] The composition of the present invention is as disclosed above in the second aspect of the invention.

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

[0142] The signs of skin ageing, and the subject in need of treatment, are as described above in the third aspect of the invention.

[0143] Furthermore, all of the embodiments discussed above in relation to the third aspect of the invention are also directly applicable to this fourth aspect of the invention, with appropriate adaptations.

[0144] In a fifth aspect, the present invention relates to a method for the prevention and / or treatment of skin ageing or inflammatory ageing in a subject, comprising administering to the subject a peptide of the invention or a composition of the invention.

[0145] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0146] The composition of the present invention is as disclosed above in the second aspect of the invention.

[0147] The composition of the present invention is preferably a cosmetic composition as disclosed above in the second aspect of the present invention.

[0148] The signs of skin ageing, inflammatory ageing and the subject are as described above in the third aspect of the invention.

[0149] Furthermore, all of the embodiments discussed above in the third and fourth aspects of the invention are directly applicable to this fifth aspect of the invention, with appropriate adaptations.

[0150] In a sixth aspect, the present invention relates to a composition or a peptide of the invention for use as a medicament.

[0151] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0152] The composition of the present invention is as disclosed above in the second aspect of the invention.

[0153] The peptides or compositions of the invention are used in a pharma- ceutical effective amount or quantity.

[0154] The composition of the present invention is preferably a pharmaceutical composition as disclosed above in the second aspect of the present invention.

[0155] In this sixth aspect of the invention, a composition or peptide of the invention is administered to a subject in need of treatment. Preferably, the subject in need of treatment is a mammal, more preferably a human.

[0156] In a preferred embodiment, the pharmaceutical use is for use in the prevention, amelioration and / or treatment of age-related diseases, inflammatory diseases and / or diseases associated with oxidative stress.

[0157] In a preferred embodiment, the pharmaceutical use is for the prevention, amelioration and / or treatment of an inflammatory disease.

[0158] Preferably, the inflammatory disease is a chronic inflammatory disease, more preferably rheumatoid arthritis, inflammatory bowel disease, psoriasis or palmoplantar pustulosis.

[0159] Also preferably, the inflammatory disease is rosacea, atopic skin, psoriasis or acne.

[0160] It is also contemplated that the inflammatory disease is inflammatory aging, more preferably non-cosmetic inflammatory aging, i.e., pathological inflammatory aging.

[0161] It is also contemplated that the inflammatory condition is inflammatory loose skin or inflammatory sagging skin (ie, pathological loose skin or pathological sagging skin).

[0162] In a further preferred embodiment the pharmaceutical use is for the prevention, amelioration and / or treatment of diseases associated with oxidative stress.

[0163] Preferably, the disease associated with oxidative stress is selected from cardiovascular disease (CVD) (preferably atherosclerosis), chronic obstructive pulmonary disease, acute or chronic renal disease, clinical frailty, neurodegenerative diseases (preferably AD, Parkinson's disease (PD), sarcopenia, Huntington's disease (HD), amyotrophic lateral sclerosis (ALS) and vascular dementia), macular degeneration (MD), biliary tract disease cancer, biliary cirrhosis, cholangitis, osteoarthritis and diabetes.

[0164] In a seventh aspect, the present invention relates to the use of a composition or a peptide of the invention for the manufacture of a medicament for the prevention, amelioration and / or treatment of an age-related disease, an inflammatory disease or a disease associated with oxidative stress.

[0165] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0166] The composition of the present invention is as disclosed above in the second aspect of the invention.

[0167] The peptides or compositions of the invention are used in a pharma- ceutical effective amount or quantity.

[0168] The composition of the present invention is preferably a pharmaceutical composition as disclosed above in the second aspect of the present invention.

[0169] The ageing-related diseases, inflammatory diseases and diseases associated with oxidative stress are as disclosed above in the sixth aspect of the present invention.

[0170] In this seventh aspect of the invention, a composition or peptide of the invention is administered to a subject in need of treatment. Preferably, the subject in need of treatment is a mammal, more preferably a human.

[0171] In an eighth aspect, the present invention relates to a method for the prevention, amelioration and / or treatment of an age-related disease, an inflammatory disease or a disease associated with oxidative stress, comprising administering to a subject in need thereof a peptide or composition of the present invention.

[0172] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0173] The composition of the present invention is as disclosed above in the second aspect of the invention.

[0174] The peptides or compositions of the invention are used in a pharma- ceutical effective amount or quantity.

[0175] The composition of the present invention is preferably a pharmaceutical composition as disclosed above in the second aspect of the present invention.

[0176] The ageing-related diseases, inflammatory diseases and diseases associated with oxidative stress are as disclosed above in the sixth aspect of the present invention.

[0177] Preferably, the subject in need of treatment is a mammal, more preferably a human.

[0178] In a ninth aspect, the present invention relates to a dietary supplement composition comprising a peptide of the present invention.

[0179] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0180] In a final aspect, the present invention relates to the use of a peptide of the invention or a composition of the invention as a dietary supplement.

[0181] The peptide of the invention is as disclosed above in the first aspect of the invention.

[0182] The composition of the present invention is as disclosed above in the second aspect of the present invention. More preferably, the composition of the present invention is a food composition as described in the second aspect of the present invention.

[0183] The activities demonstrated in the experiments described above and contained below, particularly the anti-inflammatory and antioxidant activities, indicate the utility of the peptides and compositions of the invention as dietary supplements.

[0184] [Sequence Listing Free Text] (SEQ ID NO:1) Artificially synthesized sequences Xaa is a small amino acid Xaa is selected from a polar uncharged amino acid or an aromatic amino acid Xaa is absent or selected from aromatic amino acids Xaa is selected from negatively charged amino acids Xaa is selected from non-polar amino acids Xaa is selected from Asn or Leu Xaa is selected from negatively charged amino acids Xaa is absent or selected from aromatic amino acids (SEQ ID NO:2) Artificially synthesized sequences (SEQ ID NO:3) Artificially synthesized sequences

[0185] [Example] [Abbreviation] The abbreviations used for amino acids in the text follow the recommendations of the 1983 IUPAC-IUB Joint Commission on Biochemical Nomenclature (reviewed in Eur. J. Biochem. (1984) 138:937).

[0186] Examples include: Asn, asparagine; Asp, aspartic acid; BSA, bovine serum albumin; cDNA, complementary DNA; C-terminus, carboxy terminus; DAPI, 4',6-diamidino-2-phenylindole; DCM, dichloromethane; DIPCDI, N,N'-diisopropylcarbodiimide; DMEM, Dulbecco's modified Eagle's medium; DMF, N,N-dimethylformamide; DMSO, dimethylsulfoxide; DNA, deoxyribonucleic acid; equiv, equivalent; ESI-MS, electrospray ionization mass spectrometry; FBS, fetal bovine serum; Fmoc, 9-fluorenylmethyloxycarbonyl; Glu, glutamic acid; Gly, glycine; HOBt, 1-hydroxybenzotriazole; HPLC, high performance liquid chromatography; IFN-γ, interferon gamma; Leu, leucine; LSGS, Low Serum Growth Supplement. Supplement); MBHA, p-methylbenzhydrylamine; Me, methyl; MeCN, acetonitrile; MeOH, methanol; N-terminus, amino terminus; PBS, Phosphate Borate Saline; qPCR, real-time polymerase chain reaction; RH, relative humidity; RIPA, radioimmunoprecipitation assay buffer; RNA, ribonucleic acid; RT, room temperature; RT-qPCR, quantitative reverse transcription polymerase chain reaction; tBu, tert-butyl; TFA, trifluoroacetic acid; TIS, triisopropylsilane; Trt, triphenylmethyl or trityl; Tyr, tyrosine.

[0187] For the chemical synthesis procedures included in this example, it should be noted that all synthesis processes were carried out in polypropylene syringes fitted with porous polyethylene disks or Pyrex® reactors fitted with porous plates. All reagents and solvents were of synthesis quality and were used without further treatment. Solvents and soluble reagents were removed by aspiration. Fmoc groups were removed with piperidine-DMF (2:8, volume / volume, v / v) (at least 1×1 min, 2×10 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 deprotection, coupling, and deprotection again steps were performed with DMF (3×1 min) and DCM (3×1 min) each time, using 10 mL solvent / g resin. Coupling reactions were carried out with 3 mL solvent / g resin. Coupling control was performed by carrying out the ninhydrin test (Kaiser E. et al., Anal. Biochem., 1970, 34: 595598). All synthesis reactions and washings were carried out at RT.

[0188] [Example 1] Synthesis and preparation of peptides a) Obtaining Fmoc-AA1-AA2-AA3-AA4-AA5-AA6-AA7-Rink-MBHA-resin, where AA1 is L-Gly; AA2 is L-Tyr; AA3 is L-Tyr; AA4 is L-Glu; AA5 is L-Leu; AA6 is L-Asn, and AA7 is L-Asp. Weight was normalized. 4.8 g (2.5 mmol) of Fmoc-Rink-MBHA resin with 0.52 mmol / g functionalization was treated with piperidine-DMF according to the general protocol described in the art to remove the Fmoc group. 3.08 g of Fmoc-L-Asp(tBu)-OH (7.5 mmol; 3 equiv) was loaded 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 solvent for 1 h.

[0189] The resin was then washed and the Fmoc group deprotection process was repeated to couple the next amino acid as described in common methods known in the art. Following the protocol previously described, 4.45 g of Fmoc-L-Asn(Trt)-OH (7.5 mmol; 3 equiv); followed by 2.65 g of Fmoc-L-Leu-OH (7.5 mmol; 3 equiv); followed by 3.19 g of Fmoc-L-Glu(OtBu)-OH (7.5 mmol; 3 equiv); followed by 3.44 g of Fmoc-L-Tyr(tBu)-OH (7.5 mmol; 3 equiv), followed by 3.44 g of Fmoc-L-Tyr(tBu)-OH (7.5 mmol; 3 equiv) and followed by 2.23 g of Fmoc-L-Gly-OH (7.5 mmol; 3 equiv), respectively, were coupled successively 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 mentioned above, a deprotection treatment of the Fmoc group was carried out between each amino acid addition step.

[0190] After synthesis, the peptide resin was washed with DCM (5 times for 3 min each) and dried under vacuum.

[0191] b) Obtaining Fmoc-AA1-AA2-AA3-AA4-AA5-AA6-AA7-Rink-MBHA-resin, where AA1 is L-Gly; AA2 is L-Asn; AA3 is L-Glu; AA4 is L-Leu; AA5 is L-Leu; AA6 is L-Asp, and AA7 is L-Tyr. Weights were normalized. 4.8 g (2.5 mmol) of Fmoc-Rink-MBHA resin with 0.52 mmol / g functionalization was treated with piperidine-DMF according to the general protocol described in the art to remove the Fmoc group. 3.44 g of Fmoc-L-Tyr(tBu)-OH (7.5 mmol; 3 equiv) was loaded 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 solvent for 1 h.

[0192] The resin was then washed and the Fmoc group deprotection procedure was repeated to couple the next amino acid as described in common methods known in the art. According to the protocol described above, 3.08 g of Fmoc-L-Asp(tBu)-OH (7.5 mmol; 3 equiv); followed by 2.65 g of Fmoc-L-Leu-OH (7.5 mmol; 3 equiv); followed by 2.65 g of Fmoc-L-Leu-OH (7.5 mmol; 3 equiv); followed by 3.19 g of Fmoc-L-Glu(OtBu)-OH (7.5 mmol; 3 equiv); followed by 4.45 g of Fmoc-L-Asn(Trt)-OH (7.5 mmol; 3 equiv) and followed by 2.23 g of Fmoc-L-Gly-OH (7.5 mmol; 3 equiv), respectively, were coupled consecutively 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 mentioned above, a deprotection treatment of the Fmoc group was carried out between each amino acid addition step.

[0193] After synthesis, the peptide resin was washed with DCM (5 times for 3 min each) and dried under vacuum.

[0194] Using the synthesis procedure described above and selecting the required amino acids, the following sequences were synthesized: R1-Gly-Tyr-Tyr-Glu-Leu-Asn-Asp-R2 (R1-SEQ ID NO:2-R2); or R1-Gly-Asn-Glu-Leu-Leu.Asp-Tyr-R2 (R1-SEQ ID NO:3-R2)

[0195] c) Removal of the Fmoc N-terminal protecting group of the peptide synthesized according to a) or b). The N-terminal Fmoc group of the peptidyl resin obtained in a) or b) above was deprotected with 20% (volume / volume, hereafter v / v) piperidine in DMF (1×1 min+2×10 min) (Lloyd Williams P. et al. (1997) Chemical Approaches to the Synthesis of Peptides and Proteins. CRC, Boca Raton (Fla., USA)). The peptidyl resin was washed with DMF (5×1 min) and DCM (4×1 min) and dried under vacuum.

[0196] d) The cleavage process of the peptidyl resin obtained according to c) from the polymer support. The weight was normalized. 200 mg of the dried peptidyl resin obtained in c) was treated with 5 mL of TFA / TIS / HO (volume ratio 90:5:5) for 2 h at room temperature with stirring. The filtrate was collected and precipitated using 50 mL (8-10 times) of cold diethyl ether. The ether solution was evaporated to dryness under reduced pressure at room temperature, and the precipitate was redissolved in 50% (v / v) MeCN in HO and lyophilized.

[0197] e) Characterization of peptides synthesized and prepared according to d). HPLC analysis of the peptides obtained according to d) was performed using a Shimadzu instrument (Kyoto, Japan) with a gradient of MeCN (+0.036% (v / v) TFA) in HO (+0.045% (v / v) TFA) at a flow rate of 1.25 mL / min, detection at 220 nm, using a reversed-phase column (150 × 4.6 mm, XBridge Peptide BEH C18, 3.5 μm, Waters, USA). All peptides showed a purity of more than 80%. The identity of the obtained peptides was confirmed by ESI-MS on a Water ZQ 4000 detector, using MeOH as mobile phase at a flow rate of 0.2 mL / min. The obtained results demonstrated that the peptides of sequence ID 2-NH2 and sequence ID 3-NH2 were correctly and effectively synthesized.

[0198] [Example 2] Expression analysis of inflammation-related genes For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0199] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain a final working concentration of 0.01 mg / mL in complete medium (Dermal Cell Basal Medium (DCBM) supplemented with Keratinocyte Growth Kit and 0.1% penicillin / streptomycin). Untreated cells were used as negative control samples (basal).

[0200] The relevant genes whose expression was measured are as follows: TOLLIP (toll interacting protein), IRAK3 (interleukin 1 receptor-associated kinase 3), TNFAIP3 (TNFα-induced protein 3), HMOX1 (heme oxygenase 1), NQO1 (NAD(P)H quinone dehydrogenase 1), NFKB1 (nuclear factor kappa B subunit 1), TNF (tumor necrosis factor), IL1B (interleukin 1 beta), IL6 (interleukin 6), CXCL8 (CXC motif chemokine ligand 8), MMP3 (matrix metallopeptidase 3), and TLR2 (toll-like receptor 2). The primers used for each gene were commercially available TaqMan gene expression assay probes. The experiment was performed in vitro in HEKa cells, and one concentration of peptide (0.01 mg / mL) was tested at 6 and 24 h.

[0201] Briefly, adult-derived human epidermal keratinocytes (HEKa) were seeded at a density of 300,000 cells / well in 6-well plates and maintained for 24 h under standard culture conditions (DCBM medium supplemented with Keratinocyte Proliferation Kit and 0.1% penicillin / streptomycin; 37°C, 95% relative humidity, 5% CO2). HEKa were then incubated in the presence of 0.01 mg / mL peptide for 6 or 24 h.

[0202] Cells were finally lysed and RNA extraction was performed using the Qiagen RNeasy Mini kit according to the manufacturer's instructions. Purified RNA was used to generate the corresponding cDNA by reverse transcription, which served as a template for amplification. RT-qPCR was performed using a StepOne plus Real-Time PCR instrument with the panel of TaqMan assay probes mentioned above and 2x Gene Expression Master Mix. Amplification included 40 cycles of 95°C for 15 s (denaturation) and 60°C for 1 min (annealing and extension).

[0203] The data obtained were analyzed using the ΔΔCt method, which provides expression values ​​of target genes as fold changes in treated samples compared to untreated basal samples. All samples were normalized with the relative expression of the housekeeping gene GAPDH (glyceraldehyde 3-phosphate dehydrogenase).

[0204] The analytical steps included: 1. Calculate the average Ct for each condition; 2. Calculate ΔCT test sample and ΔCT calibrator sample; 3. Calculate ΔΔCT: ΔΔCT = ΔCT test sample - ΔCT calibrator sample; 4.2 -ΔΔCT The ratio is calculated by:

[0205] Gene regulation with ratios (fold change vs. basal sample) >1.15 or <(-1.15) is considered biologically significant.

[0206] The results obtained are summarized in Figures 1 and 2. These figures show that the peptide of SEQ ID NO: 2-NH2 (as an example of a peptide of the invention) is able to modulate the expression of inflammation-related genes, with anti-inflammatory genes being upregulated and inflammatory genes being downregulated. More precisely, after 6 hours, HEKa cells treated with the peptide of the invention showed an upregulation of the genes TNFAIP3, TOLLP, HMOX1 and IRAK3, while after 24 hours of treatment with the peptide of the invention, NQO1 was upregulated and NFKB1, IL1B, TNF, CXCL8, IL6, MMP3 and TLR2 genes were downregulated. All these results confirm or demonstrate the usefulness of the peptide of the invention against inflammation and therefore for the prevention, reduction and / or treatment of inflammatory aging and aging of the skin. Moreover, all these results also demonstrate the usefulness of the peptide of the invention for the treatment of age-related and inflammatory diseases, as explained above. This anti-inflammatory activity also demonstrates the usefulness of the peptides of the invention as dietary supplements.

[0207] [Example 3] Expression analysis of inflammation-related miRNA For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0208] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain a final working concentration of 0.01 mg / mL in complete medium (Dermal Cell Basal Medium (DCBM) supplemented with Keratinocyte Growth Kit and 0.1% Penicillin / Streptomycin). Untreated cells were used as negative control samples (Basal).

[0209] The relevant miRNAs whose expression was measured were miR-21-5p and miR-146a-5p. In the last few years, microRNAs (miRNAs) have been identified as another important component in aging and inflammatory aging. miR-21-5p and miR-146a-5p are involved in the regulation of the NF-κB and NLRP inflammatory pathways, and their abundance is significantly altered in both tissues and the bloodstream during aging and is associated with various inflammatory conditions. Decreased expression of this miRNA is associated with aging and has been proposed as a biomarker for age-related diseases.

[0210] The primers used for each gene were commercially available TaqMan Advanced miRNA assay probes. Experiments were performed in vitro in HEKa cells, and one concentration of peptide (0.01 mg / mL) was tested for 24 h.

[0211] Briefly, adult-derived human epidermal keratinocytes (HEKa) were seeded at a density of 300,000 cells / well in 6-well plates and maintained for 24 h under standard culture conditions (DCBM medium supplemented with Keratinocyte Proliferation Kit and 0.1% penicillin / streptomycin; 37°C, 95% relative humidity, 5% CO2). HEKa were then incubated in the presence of 0.01 mg / mL of peptide for 24 h.

[0212] Cells were finally lysed and RNA extraction was performed using the miRVana miRNA isolation kit according to the manufacturer's instructions. Purified miRNA was used to generate the corresponding cDNA by reverse transcription, which served as a template for amplification. RT-qPCR was performed using a StepOne plus Real-Time PCR instrument with the TaqMan Advanced miRNA assay probe panel described above and the Taqman Fast Advance master mix. Amplification included 40 cycles of 95°C for 1 s (denaturation) and 60°C for 20 s (annealing and extension).

[0213] The data obtained were analyzed using the ΔΔCt method, which provides expression values ​​of target genes as fold changes in treated samples compared to untreated basal samples. All samples were normalized to the relative expression of the housekeeping miRNA: miR-16-5p.

[0214] The analytical steps included: 1. Calculate the average Ct for each condition; 2. Calculate ΔCT test sample and ΔCT calibrator sample; 3. Calculate ΔΔCT ΔΔCT = ΔCT test sample - ΔCT calibrator sample; 4.2 -ΔΔCT The ratio is calculated by:

[0215] Three independent experiments (N=3) were performed.

[0216] The results obtained are summarized in FIG. 3. This figure shows that the peptide of SEQ ID NO: 2-NH2 (as an example of a peptide of the invention) was able to regulate the expression of miRNAs with anti-inflammatory properties. More precisely, the peptide of the invention was able to upregulate the expression of both miR-146a-5p and miR-21-5p. All these results confirm or demonstrate the usefulness of the peptide of the invention against inflammation and therefore for the prevention, reduction and / or treatment of inflammatory aging and skin aging. Moreover, all these results also demonstrate the usefulness of the peptide of the invention for the treatment of age-related and inflammatory diseases, as explained above. This anti-inflammatory activity also demonstrates the usefulness of the peptide of the invention as a dietary supplement.

[0217] [Example 4] Analysis of IL-8 levels For this example, peptides SEQ ID NO:2-NH2 and SEQ ID NO:3-NH2 were prepared according to Example 1.

[0218] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain final working concentrations of 0.005, 0.01 and 0.05 mg / mL in complete medium (DCBM supplemented with Keratinocyte Proliferation Kit and 0.1% penicillin / streptomycin). Cells treated with 10 ng / mL IFNγ alone were used as a positive control for inflammation induction. Untreated cells were used as basal conditions.

[0219] In this example, HaCaT keratinocytes were seeded in 96-well plates at a density of 75,000 cells / well and maintained under standard culture conditions (DMEM with 10% FBS and 1% penicillin / streptomycin; 37°C, 95% RH, 5% CO2) for 24 hours. Keratinocytes were then treated with the peptides of the invention at the above-mentioned concentrations in the presence of 10 ng / mL IFNγ in DMEM + 1% FBS for 48 hours. After treatment, the supernatants were collected, centrifuged to remove cell debris, and stored at -80°C until used for IL-8 quantification using the Human IL-8 / CXCL8 DuoSet ELISA Kit according to the manufacturer's instructions.

[0220] To determine cell viability, at the end of the experiment, cells were incubated with 0.5 mg / mL MTT in complete medium for 1 h, then the MTT solution was removed and the formazan crystals formed were dissolved with 150 μL DMSO / well. Finally, the OD at 570 nm was determined using a Multiskan spectrophotometer.

[0221] Each condition was performed in triplicate (n=3) in three independent experiments (N=3). All data obtained were normalized according to formulas (II) and (III).

number

number

[0222] The results obtained are summarized in Figures 4 and 5. These figures show that SEQ ID NO:2-NH2 and SEQ ID NO:3-NH2 (as an example of a peptide of the invention) were able to dose-dependently reduce IL-8 secretion in all conditions tested (48 hours of treatment, 0.005, 0.01 or 0.05 mg / mL, together with 10 ng / mL IFNγ) compared to the positive control (inflammation induced with 10 ng / mL IFNγ).

[0223] Similar experiments were carried out under basal conditions (i.e. without inflammatory stimulation of IFNγ) for both peptides SEQ ID NO:2-NH2 (at concentrations of 0.005, 0.01 and 0.05 mg / mL) and SEQ ID NO:3-NH2 (at concentrations of 0.01 and 0.05 mg / mL). In this case, basal or untreated cells were set as 100% (IL-8 secreted from basal or untreated cells). The results obtained are summarized in Figures 6 and 7. As in the previous case, all tested concentrations of SEQ ID NO:2-NH2 dose-dependently reduced the secretion of IL-8, but to a lesser extent than under inflammatory conditions. Moreover, surprisingly, all tested concentrations of SEQ ID NO:3-NH2 also reduced the secretion of IL-8, but to a greater extent than SEQ ID NO:2-NH2.

[0224] All the results obtained in this example demonstrate the protective effect of the peptide of the present invention, since IL-8 is an inflammatory cytokine, and therefore the peptide of the present invention is useful for preventing, reducing and / or treating inflammatory aging and skin aging.Furthermore, all these results also demonstrate that the peptide of the present invention is useful for treating aging-related and inflammatory diseases, as explained above.This anti-inflammatory activity also demonstrates the usefulness of the peptide of the present invention as a dietary supplement.

[0225] [Example 5] Analysis of telomere length For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0226] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain a final working concentration of 0.005 or 0.01 mg / mL in complete medium (Medium 106 supplemented with 2% LSGS). 73 (73-year-old adult HDFa) was used as a negative control, and untreated HDFa 25 (HDFa in a 25-year-old adult) was used for comparison purposes.

[0227] Briefly, adult-derived human dermal fibroblasts (HDFa) were seeded in 12-well plates at a concentration of 150,000 cells / well and maintained under standard culture conditions (medium 106 supplemented with 2% LSGS, 37°C, 95% RH, 5% CO2) for 24 h.

[0228] Next, HDFa cells were treated with the above peptides at the above concentrations for 48 hours. DNA was then extracted using the Qiagen DNeasy Mini kit according to the manufacturer's instructions. Finally, the telomere DNA to nuclear DNA ratio was set using the Absolute Human Telomere Length Quantitative qPCR Assay Kit according to the manufacturer's instructions. The resulting data was analyzed using the ΔΔCt method to provide the relative telomere length of the target sample as the fold change in telomere sequence (telDNA) compared to the single copy nuclear reference DNA sequence (SCR).

[0229] The analytical steps included: 1. Calculate the Ct for each condition of telDNA and SCR; 2. Calculate ΔCT of telDNA (Ct telDNA target sample-Ct telDNA reference sample) and ΔCT SCR DNA (Ct SCR target sample-Ct SCR reference sample); 3. Calculate ΔΔCT: ΔΔCT = ΔCT tel DNA - ΔCT SCR DNA; 4.2 -ΔΔCT Obtain relative telomere length by; 5. Telomere length (%) = (relative telomere length of target sample / HDFa 25 The relative telomere length of the sample is multiplied by 100.

[0230] Three independent experiments (N=3) were performed.

[0231] The results obtained are summarized in Figure 8. 73 is HDFa 25However, HDFa incubated with 0.005 or 0.01 mg / mL of the peptide of SEQ ID NO:2-NH2 (as an example of a peptide of the present invention) for 48 hours showed telomeres that were 20% shorter than those of HDFa 73 are untreated HDFa, 73 The HDFa showed 17% and 30% longer telomeres compared to the HDFa. Thus, the peptides of the present invention can induce telomere length restoration and 73 Telomere length of HDFa 25 The study was able to restore telomere length to levels similar to those observed in humans, demonstrating utility for preventing, reducing and / or treating inflammatory aging and skin aging, as well as age-related diseases, inflammatory diseases and diseases associated with oxidative stress.

[0232] [Example 6] Analysis of intracellular localization of Nrf2 For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0233] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain a final working concentration of 0.01 or 0.05 mg / mL in complete medium (Keratinocyte Growth Kit and Skin Basal Medium supplemented with 0.1% penicillin / streptomycin). Untreated cells were used as negative control samples (basal conditions).

[0234] In this study, HEKa cells were cultured at a density of 50,000 cells / well in skin cell basal medium (DCBM) and incubated at 37°C in a humidified atmosphere of 5% CO2. When the cells reached approximately 80% confluency, they were treated with various concentrations of the peptides listed above for 24 hours. After treatment, the samples were washed twice with PBS, fixed with 4% formaldehyde, and permeabilized with 0.2% Triton X-100. The samples were then blocked with 5% PBS / BSA for 1 hour to avoid nonspecific antibody binding.

[0235] For Nrf2 staining, samples were incubated with 1:100 Alexa Fluor® 488 anti-NRF2 antibody in 1% PBS / BSA overnight at 4°C. After appropriate washing with PBS, cells were incubated with 50 μg / mL Phalloidin-TRITC (for actin filament staining) for 1 h at room temperature and in the dark. Finally, samples were washed three times with PBS and coverslip mounted with Prolong-Gold containing DAPI, which strongly binds to DNA (nuclear stain). Microscopic images were acquired using a 10x objective. Immunofluorescence was replicated for each condition, and images of at least five different fields from each coverslip were acquired with the same settings.

[0236] The images obtained were analyzed by ImageJ software for fluorescence intensity in the nucleus (DAPI staining) and cytoplasm (Nrf2 staining).

[0237] The percentage of Nrf2 located in the nucleus was obtained by normalizing the nuclear fluorescence signal obtained for Nrf2 in treated samples with respect to that compared for untreated samples.

[0238] The results obtained are summarized in FIG. 9. It can be seen that the peptide of SEQ ID NO: 2-NH2 (as an example of the peptide of the present invention) can provide Nrf2 translocation from the cytoplasm to the nucleus in a concentration-dependent manner. As is widely known, in the nucleus, Nrf2 in the nucleus induces the expression of various antioxidant genes. Thus, the peptide of the present invention can provide anti-inflammatory and antioxidant activity, and is useful for preventing, reducing and / or treating inflammatory aging and skin aging. Furthermore, all these results also demonstrate that the peptide of the present invention is useful for treating aging-related diseases, inflammatory diseases and diseases related to oxidative stress, as explained above. This anti-inflammatory and antioxidant activity also demonstrates the usefulness of the peptide of the present invention as a dietary supplement.

[0239] [Example 7] Analysis of TLR2 levels in human epidermal keratinocytes For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0240] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain a final working concentration of 0.01 or 0.05 mg / mL in complete medium (Keratinocyte Proliferation Kit and Skin Basal Medium supplemented with 0.1% penicillin / streptomycin). Cells treated with 10 ng / mL IFNγ alone were used as a positive control sample for inflammation induction. Untreated cells were used as a negative control sample (basal condition).

[0241] In this study, HEKa cells were seeded in 24-well plates at a density of 50,000 cells / well and maintained for 24 hours under standard conditions (DCBM supplemented with Keratinocyte Proliferation Kit and 0.1% penicillin / streptomycin; 37°C, 95% RH, 5% CO2). Keratinocytes were then treated with 10 ng / mL IFNγ for 24 hours. IFNγ was then removed and cells were incubated with various concentrations of the peptides of the invention as described above for 24 hours. During this time, positive control samples (IFNγ only) were incubated with complete medium without peptide. After treatment, cells were washed with Dubelcco PBS (PBS also containing potassium chloride) and lysed with RIPA and scrapper. Lysates were collected, centrifuged to remove cell debris, and protein concentrates were stored at -80°C until TLR2 was quantified using the Human TLR2 DuoSet ELISA Kit according to the manufacturer's instructions.

[0242] Each condition was performed in triplicate (n=3) in three independent experiments (N=3).

[0243] The results are summarized in Figure 10, and show that the peptide of SEQ ID NO: 2-NH2 (as an example of the peptide of the present invention) can dose-dependently reduce the amount of TLR2 after exposure to IFNγ, with a maximum reduction of 19% after treatment with the highest peptide concentration. Thus, the peptide of the present invention has a protective role against skin inflammation and is useful for preventing, reducing and / or treating inflammatory aging and skin aging. Furthermore, all these results also demonstrate that the peptide of the present invention is useful for treating age-related and inflammatory diseases, as explained above. This anti-inflammatory activity also demonstrates the usefulness of the peptide of the present invention as a dietary supplement.

[0244] [Example 8] Analysis of the amount or synthesis of catalase enzyme For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0245] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO. This stock solution was further diluted to obtain final working concentrations of 0.005, 0.01 or 0.05 mg / mL in complete medium (Keratinocyte Growth Kit and Skin Basal Medium supplemented with 0.1% penicillin / streptomycin). Untreated cells were used as negative control samples (basal conditions).

[0246] In this study, HEKa cells were seeded in 24-well plates at a density of 50,000 cells / well and maintained in standard culture conditions (DCBM; 37°C, 95% RH, 5% CO2) for 24 hours. Keratinocytes were then treated with the peptides of the invention at the above-mentioned concentrations for 24 hours. After treatment, cells were washed with Dubelcco PBS and lysed with RIPA and scrapper. Lysates were collected, centrifuged to remove cell debris, and protein concentrates were stored at -80°C until used for catalase quantification using a human catalase ELISA kit according to the manufacturer's instructions. Finally, to normalize the data, the total protein content of each sample was quantified with a Micro BCA protein assay kit according to the manufacturer's instructions.

[0247] Each condition was performed at least in triplicate (n=3) in three independent experiments (N=3).

[0248] The results obtained are summarized in Figure 11, where it can be seen that the peptide of SEQ ID NO: 2-NH2 (as an example of a peptide of the invention) is able to increase the amount of catalase in a dose-dependent manner, with an increase of 47% at the highest concentration. These results demonstrate the ability of the peptide of the invention to provide an antioxidant response, thus demonstrating its usefulness for the prevention, reduction and / or treatment of inflammatory aging and skin aging. This antioxidant activity also demonstrates the usefulness of the peptide of the invention in the treatment of diseases related to oxidative stress (as explained above) and as a dietary supplement.

[0249] [Example 9] Analysis of TRF1 quantity For this example, peptides SEQ ID NO:2-NH2 and SEQ ID NO:3-NH2 were prepared according to Example 1.

[0250] It is widely known in the art that in vertebrates, telomeres are composed of tandem repeats of the TTAGGG sequence and are bound by six protein complexes known as shelterins, including TRF1, TRF2, TIN2, POT1, RAP1 and TPP1. Telomere repeat binding factor 1 (TRF1) was the first shelterin discovered (Zhong, Z., et al. (1992) A mammalian factor that binds telomeric TTAGGG repeats in vitro. Mol Cell Biol 12, 4834-4843.) and is one of the best characterized factors. TRF1 binds to double-stranded TTAGGG repeats, its abundance at telomeres is proportional to the length of the repeat (de Lange, T. (2005) Shelterin: the protein complex that shapes and safeguards human telomeres. Genes. Dev. 19, 2100-2110, and Liu, L., et al. (2007) Telomere lengthening early in development. Nat. Cell Biol. 9:1436-1441), making it an excellent reporter of telomere length in vivo.

[0251] CHA9.3 cells were seeded at 10,000 cells per well in a sterile 96-well black-clear bottom tissue culture plate using 150 μL of DMEM medium supplemented with 10% fetal bovine serum. Cells were incubated at 37°C with 5% CO2 in humidified air for 24 hours.

[0252] In parallel, the peptides used in this example were weighed and diluted in dimethyl sulfoxide (DMSO) or water to generate a "mother" plate. From this, intermediate dilution plates were prepared. 1.5 μL of each compound solution was automatically dispensed from the intermediate dilution plate into the medium of cells grown in culture medium using a Biomek FX Dual Pod automated liquid handling robot to obtain final concentrations of 0.125 mg / mL (peptide with SEQ ID NO:2-NH2), 0.063 mg / mL (peptide with SEQ ID NO:3-NH2). Each compound was biochemically analyzed in duplicate. The final concentration for screening was determined based on a viability assay. Each plate also contained water or DMSO controls as basal conditions.

[0253] After 24 h of incubation at 37 °C in humidified air with 5% CO2, cells were processed for immunofluorescence. Briefly, cells were fixed with 4% formaldehyde in phosphate-buffered saline (PBS) for 13 min and washed twice again with PBS at room temperature. Cells were then permeabilized with 0.2% Triton X-100 detergent for 10 min, washed again with PBS, and then permeabilized with 0.1% sodium citrate + 0.1% Triton X-100 for 5 min, followed by a final wash to remove residual buffer. Cells were blocked with 0.05% PBS-Tween-20 at 50 μM and 3% bovine serum albumin for 60 min at room temperature. Immunostaining was performed by incubation with anti-TRF1 primary antibody overnight at 4 °C. Afterwards, the cells were washed three times with 0.05% PBS-Tween-20 and the secondary antibody anti-rat-555 was dispensed at 1:500 in PBS-Tween + 3% BSA for 60 min. Samples were then washed three times with 0.05% PBS-Tween-20 and further washed with PBS. Finally, they were incubated with 1 μg / mL Hoechst stain and a mask was created corresponding to the cell nuclei. SPSS software was used for statistical analysis. Image acquisition was performed on an Opera™ LX spinning disk confocal automation platform (PerkinElmer). Forty images were acquired with a 40x magnification lens under non-saturating conditions, and an average of 1 × 10 3Quantification of fluorescence at the single cell level was performed on the Acapella™ platform using built-in analysis algorithms to identify aggregate and nucleus area and intensity.

[0254] Cell viability in each condition was also assessed by comparing nuclei counts with controls.

[0255] Each condition was performed at least in duplicate (n=2) in three independent experiments (N=3). For all data, the mean TRF1 intensity value of the different experiments was calculated.

[0256] The results obtained are summarized and presented in Figure 12. As can be directly derived from this figure, the peptides of the invention (peptides of SEQ ID NO:2-NH2 and SEQ ID NO:3-NH2) provide an increase in TRF1, which, as mentioned above, is directly related to telomere length (increased telomere length), demonstrating the usefulness of the peptides of the invention for preventing, reducing and / or treating inflammatory aging and skin aging, as well as age-related diseases, inflammatory diseases and diseases related to oxidative stress.

[0257] [Example 10] Analysis of TRF1 and DNA damage under oxidative stress conditions For this example, the peptide of SEQ ID NO:2-NH2 was prepared according to Example 1.

[0258] Standard stock solutions of peptides were prepared at 50 mg / mL in DMSO, which were further diluted to give final working concentrations of 0.005 mg / mL, 0.01 mg / mL and 0.05 mg / mL pure activity.

[0259] HEKa cells were seeded on cover slips in 24-well plates containing skin cell basal medium. Cultures were then maintained in a 37°C incubator to allow attachment and proliferation. When cells were approximately 50% confluent, the medium was replaced with medium containing the various concentrations of peptides to be tested (5, 10 or 50 μg / mL of peptide of sequence number 2-NH2) for 24 hours. The medium was then replaced with medium containing compound (for wells in basal condition experiments) or medium containing 100 μM H2O2 and compound (for wells in oxidative stress conditions) for another 24 hours.

[0260] After treatment, cells were harvested, washed once with PBS, treated with Triton X-100 buffer for 30 s for nuclear extraction, washed again with PBS, fixed with 4% buffered formaldehyde for 10 min, washed with PBS, and stored at 4°C until processing.

[0261] For immunofluorescence (IF), cells were permeabilized with 0.2% PBS-Triton for 10 min, washed, and blocked with fetal bovine serum for 1 h and with 5% BSA in PBS for an additional 1 h to avoid nonspecific antibody binding.

[0262] For staining of TRF1 and 53BP1, the cells were incubated with the corresponding primary antibodies overnight at 4 °C. After appropriate washing with PBS, the cells were incubated with 488-Alexa secondary antibody for 1 h in a humid chamber. The samples in the cover slides were mounted with Prolong Gold containing DAPI, which strongly binds to DNA (nuclear stain), and the samples were protected from light at 4 °C until microscopic images were acquired. The fluorescent signals on the slides were visualized with a confocal ultraspectromicroscope SP5-MP (Leica). Microscopic images were acquired using a 40x objective. Immunofluorescence was performed in triplicate for each condition, and images of at least four different fields from each cover slip were acquired with the same settings.

[0263] Signals were quantified using Definiens Developer XD.2 software. Image analysis was performed blinded. The blue channel (DAPI staining) was used to count the number of nuclei per image. Red and green were used to determine the intensity of TRF1 and 53BP1, respectively. The number of TRF1 or 53BP1 aggregates was then divided by the number of nuclei to determine the total number of TRF1 or 53BP1 aggregates / nucleus for each image. Finally, all data were normalized to negative controls (basal samples for TRF1 and H2O2-treated samples for 53BP1) to determine the percentage (%) of TRF1 or 53BP1 aggregates per nucleus.

[0264] The results obtained for TRF1 are summarized and presented in Figure 13. As can be easily seen from the figure, the peptides of the present invention (exemplified by the peptide of SEQ ID NO:2-NH2) provide an increase in TRF1, which, as described above, is directly related to telomere length (increased telomere length), demonstrating the usefulness of the peptides of the present invention for preventing, reducing and / or treating inflammatory aging and dermal aging.

[0265] The results obtained for 53BP1 are summarized and presented in Figure 14. As can be easily seen from the figure, the peptide of the present invention (exemplified by the peptide of SEQ ID NO: 2-NH2) provides a reduction in 53BP1 aggregates, i.e., a reduction in 53BP1, which in turn shows a reduction in the number of double-strand breaks. This demonstrates the peptide's ability to protect cells from DNA damage induced by oxidative stress. These results therefore demonstrate the usefulness of the peptide of the present invention for preventing, reducing and / or treating inflammatory aging and skin aging. These results also demonstrate the antioxidant potential of the peptide of the present invention, and therefore demonstrate its usefulness in treating aging-related diseases and diseases related to oxidative stress, as well as dietary supplements. [Brief description of the drawings]

[0266] [Figure 1]1 shows the gene expression results obtained in Example 2 when HEKa cells were treated with 0.01 mg / mL of the peptide of SEQ ID NO:2-NH2 for 6 hours. [Diagram 2] 1 shows the gene expression results obtained in Example 2 when HEKa cells were treated with 0.01 mg / mL of the peptide of SEQ ID NO:2-NH2 for 24 hours. [Diagram 3] 1 shows the miRNA expression results obtained in Example 3 when HEKa cells were treated with 0.01 mg / mL of the peptide of SEQ ID NO:2-NH2 for 24 hours. [Figure 4] In Example 4, the results obtained for the secretion or release of IL-8 under inflammatory conditions (excluding untreated or basal samples) when HaCaT keratinocytes were treated with the peptide of SEQ ID NO: 2-NH2 are shown. [Diagram 5] In Example 4, the results obtained for the secretion or release of IL-8 under inflammatory conditions (excluding untreated or basal samples) when HaCaT keratinocytes were treated with the peptide of SEQ ID NO: 3-NH2 are shown. [Figure 6] In Example 4, the results obtained on the secretion or release of IL-8 under non-inflammatory conditions when HaCaT keratinocytes were treated with the peptide of SEQ ID NO: 2-NH2 are shown. [Figure 7] In Example 4, the results obtained on the secretion or release of IL-8 under non-inflammatory conditions when HaCaT keratinocytes were treated with the peptide of SEQ ID NO: 3-NH2 are shown. [Figure 8] In Example 5, the results obtained regarding telomere length when HDFa cells were treated with the peptide of SEQ ID NO:2-NH2 are shown. [Figure 9] Example 6 shows the results obtained regarding Nrf2 translocation into the nucleus when HEKa cells were treated with the peptide of SEQ ID NO:2-NH2. [Figure 10] In Example 7, the results obtained regarding the amount of TLR2 when HEKa cells were treated with the peptide of SEQ ID NO:2-NH2 are shown. [Figure 11]In Example 8, the results obtained on the amount or synthesis of catalase when HEKa cells were treated with the peptide of SEQ ID NO:2-NH2 are shown. [Figure 12] In Example 9, the results obtained for TRF1 production (TRF1 amount measured by the measured intensity) are shown when CHA9.3 cells are treated with the peptides SEQ ID NO:2-NH2 and SEQ ID NO:3-NH2. [Figure 13] In Example 10, the results obtained for TRF1 foci per cell are shown when HEKa cells were treated with the peptide of SEQ ID NO:2-NH2. [Figure 14] 1 shows the results obtained for DNA protection (measured based on 53BP1 aggregates per cell) obtained in Example 10, where HEKa cells were treated with the peptide of SEQ ID NO:2-NH2.

Claims

1. Peptides of formula (I): <h2 style=";text-align:left;direction:ltr">1<h2 style=";text-align:left;direction:ltr"> １ <h2 style=";text-align:left;direction:ltr"> I am sorry, I did not find this information.<h2 style=";text-align:left;direction:ltr"> ２ (I) and its acceptable isomers, salts, solvates, and / or mixtures thereof, AA1 is selected from Cys, Val, Ala, Ser, Asn, Ala, Gly, or Thr; AA2 is selected from Tyr, Ser, Thr, Gln, Asn, Cys, or Gly; AA3 is absent or selected from Tyr, Phe, or Trp; AA4 is selected from Asp or Glu; AA5 is selected from Val, Ile, Leu, Met, Pro, Phe or Trp; AA6 is selected from Asn or Leu; AA7 is selected from Asp or Glu; AA8 is absent or selected from Tyr, Phe, or Trp; R １ is H, substituted or unsubstituted acyclic aliphatic, substituted or unsubstituted alicyclyl, substituted or unsubstituted heterocyclyl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted aralkyl, and R ５ —CO—, R ５ is a substituted or unsubstituted C １ ~C ２４ Alkyl radical, substituted or unsubstituted C ２ ~C ２４ Alkenyl, substituted or unsubstituted C ２ ~C ２４ Alkynyl, substituted or unsubstituted C ３ ~C ２４ Cycloalkyl, substituted or unsubstituted C ５ ~C ２４ Cycloalkenyl, substituted or unsubstituted C ８ ~C ２４ Cycloalkynyl, substituted or unsubstituted C ６ ~C ３０ Aryl, substituted or unsubstituted C ７ ~C ２４ is selected from the group consisting of aralkyl, a substituted or unsubstituted 3- to 10-membered heterocyclyl ring, and a substituted or unsubstituted heteroarylalkyl of 2 to 24 carbon atoms, 1 to 3 non-carbon atoms, and an alkyl chain of 1 to 6 carbon atoms; and R ２ is H, -NR ３ R ４ -, -OR ３ and -SR ３ where R ３ and R ４ is independently selected from H, a substituted or unsubstituted acyclic aliphatic group, a substituted or unsubstituted alicyclyl, a substituted or unsubstituted heterocyclyl, a substituted or unsubstituted heteroarylalkyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted aralkyl; Or, R １ and R ２ is absent and AA1 is directly linked to the last amino acid of the peptide, i.e., AA8 or AA7 if AA8 is absent (forming a cyclic peptide).

2. In formula (I): AA1 is selected from Ser, Cys, Ala, or Gly; AA2 is selected from Tyr, Ser, Gln or Asn; AA3 is absent or selected from Tyr, Phe, or Trp; AA4 is selected from Asp or Glu; AA5 is selected from Val, He, or Leu; AA6 is selected from Asn or Leu; AA7 is selected from Asp or Glu; AA8 is absent or selected from Tyr, Phe or Trp; The peptide according to claim 1 .

3. In formula (I): AA1 is Gly; AA2 is selected from Tyr or Asn; AA3 is absent or Tyr; AA4 is GIu; AA5 is Leu; AA6 is selected from Asn or Leu; AA7 is Asp; AA8 is absent or is Tyr; 3. A peptide according to claim 1 or 2, characterized in that it is

4. The peptide is R １ -Gly-Tyr-Tyr-Glu-Leu-Asn-Asp-R ２ (R １ -SEQ ID NO:2-R ２ );or R １ -Gly-Asn-Glu-Leu-Leu-Asp-Tyr-R ２ (R １ -SEQ ID NO:3-R ２ ) The peptide according to any one of claims 1 to 3, characterized in that it is selected from the group consisting of

5. R １ The peptide according to any one of claims 1 to 4, characterized in that is H.

6. R ２ NH ２ The peptide according to any one of claims 1 to 5, characterized in that it is

7. A composition comprising a peptide according to any one of claims 1 to 6.

8. Use of a peptide according to any one of claims 1 to 6 or a composition according to claim 7 as a cosmetic in a subject in need thereof.

9. To prevent, reduce and / or treat skin aging; and / or To prevent, reduce and / or treat inflammaging; and / or To improve skin firmness and / or elasticity 9. The cosmetic use according to claim 8, characterized in that

10. 10. Use of a cosmetic product according to claim 9, characterized in that the signs of skin ageing are wrinkles, rough skin, loose skin or sagging skin.

11. A composition according to claim 7 or a peptide according to any one of claims 1 to 6 for use as a medicament.

12. A composition according to claim 7 or a peptide according to any one of claims 1 to 6 for use in the prevention, amelioration and / or treatment of an age-related disease, an inflammatory disease or a disease associated with oxidative stress.

13. Use of a peptide according to any one of claims 1 to 6 or a composition according to claim 7 as a dietary supplement.

14. A dietary supplement comprising a peptide according to any one of claims 1 to 6.

Images