Cosmetic and / or pharmaceutical compositions comprising synthetic or plant extracts for increasing expression of mitochondrial-derived peptides

Abstract

The present invention relates to a cosmetic or pharmaceutical composition comprising a physiologically acceptable medium and an active ingredient capable of increasing the expression of a mitochondrial-derived peptide, more particularly a cerebrasin, where the active ingredient may be a peptide, a plant extract, a plant extract, or a plant extract. The present invention relates to an aqueous extract of an aqueous extract, such as an aqueous extract of a fruit pulp of phoenix dactylifera, an aqueous extract of a seed of germinated red clover, an aqueous extract of a seed of Chenopodium quinoa, or an aqueous extract of a root of Iris pallida, for example, an aqueous extract of a fruit pulp of phoenix dactylifera, an aqueous extract of a seed of germinated red clover, an aqueous extract of a seed of Chenopodium quinoa, or an aqueous extract of a root of Iris pallida. The invention also relates to a cosmetic treatment method for protecting the skin and accessories from external infringement and against skin aging.

Description

Technical Field

[0001] This invention belongs to the fields of cosmetics and pharmaceuticals, and more specifically relates to plant extracts and peptides that can increase the expression of mitochondrial-derived peptides, and their use in cosmetic applications for mitochondrial biogenesis and skin lifespan. Background Technology

[0002] Mitochondria have been extensively studied in the field of aging due to their roles in cellular energy production, calcium homeostasis, apoptosis, and cell signaling. Mitochondria possess less efficient DNA repair mechanisms than those found in the cell nucleus. These inefficient DNA repair mechanisms, combined with proximity to the electron transport chain, generate large amounts of reactive oxygen species (ROS), promoting numerous mtDNA mutations throughout a person's life.

[0003] Small open reading frames (ORFs) in circular mt DNA can encode a variety of microproteins called mitochondrial-derived peptides (MDPs). Some of these peptides function intracellularly, while others are present in systemic circulation and target various tissues. Currently, eight MDPs have been identified: humanin, MOTS-c, and SHLP (small humanin-like peptides) 1–6 (Miller et al. Mitochondria-derived peptides in aging and healthspan. J Clin Invest, 2022, 132(9)).

[0004] Brain protectant (HN) is a 24-amino acid peptide encoded by the MT-RNR2 gene in the mitochondrial 16S rRNA region.

[0005] HN mediates mitochondrial state and cell survival through intracellular mechanisms or as a secretory factor via extracellular signals. Intracellularly, it binds to Bcl2-associated X protein (BAX), Bim, tBid, and IGFBP3 to inhibit caspase activity and apoptosis.

[0006] Circulating neuroprotectin levels decreased with age in the plasma of mice, monkeys, and humans. Interestingly, this was not observed in long-lived naked mole rats, a model of negligible aging and healthy aging. Human offspring of centenarians with a greater chance of living to 100 years old even showed elevated circulating neuroprotectin levels compared to age-matched controls without a particularly long-lived family history. Using a model of *C. elegans* overexpressing HN, HN has been reported to increase lifespan by regulating Foxo (Yen et al. The mitochondrial derived peptide humanin is an agulator of lifespan and healthspan. Aging 2020, Vol 12, N°12).

[0007] HN possesses anti-aging activity because it has been shown to exacerbate the aging-associated secretory phenotype (SASP) in senescent cells by stimulating the secretion of IL-6, IL-1β, IL-8, IL-10, and tumor necrosis factor α. Nephroticin protects cells in vitro from oxidative stress, serum starvation, hypoxia, and other damage (Mendelsohn and Larrick. Mitochondrial-Derived Peptides Exacerbate Senescence. Rejuvenation Res, Aug;21(4):369-373.2018).

[0008] MOTS-c is a 16-amino acid mitochondrial-derived peptide encoded by the 12S rRNA region of the mitochondrial genome. Under stress, MOTS-c translocates to the cell nucleus, where it regulates a wide range of genes in response to metabolic dysfunction. It co-localizes to mitochondria in various problems and is found in plasma, but its levels decline with age. Due to its important cellular functions and potential hormonal effects, MOTS-c has been shown to have beneficial effects on age-related diseases, and it may extend lifespan by promoting cellular homeostasis.

[0009] Small neuroprotectin-like peptides 1 to 6 (SHLPs 1–6) are encoded by a 16S rRNA region and share some biological characteristics with neuroprotectins. For example, SHLP2 protects cells from amyloid-β-induced toxicity and age-related macular degeneration, and administration of SHLP2 and SHLP3 promotes mitochondrial biogenesis, reduces ROS, and decreases mtDNA oxidation. SHLP2 may also affect lifespan and health span, similar to other MDPs.

[0010] The presence of mitochondrial-derived peptides (including neuroprotectin) was identified in primary dermal fibroblasts (Kim et al. Mitochondrial peptides modulate mitochondrial function during cellular senescence, Aging, 2018, Vol.10, No. 6).

[0011] Compounds that stimulate mitochondrial-derived peptides and exhibit activity related to mitochondrial metabolism and cellular energy levels are crucial for improving skin lifespan and preventing or treating signs of skin aging. They represent significant interest in medical research and cosmetics because they can mitigate skin and hair damage caused by external aggressors such as UV radiation, exposure to toxins, or pollutants. To the inventors' knowledge, no cosmetics have been described that include plant extracts or peptides capable of activating mitochondrial-derived peptides. Attached Figure Description

[0012] The invention and its advantages will be better understood through the following description and non-limiting embodiments with reference to the accompanying drawings, in which:

[0013] [ Figure 1 The expression of neuroprotectin in normal human skin biopsies treated with peptides SEQ ID No. 1 and SEQ ID No. 2 at 5 mg / Kg was quantified by immunofluorescence detection.

[0014] [ Figure 2 Collagen I expression in normal human skin biopsies treated with 5 mg / Kg of SEQ ID No. 1 and SEQ ID No. 2 was quantified by immunofluorescence detection.

[0015] [ Figure 3 Immunofluorescence assay was performed on dates treated with 1% ( Phoenix dactylifera The expression of neuroprotectin in normal human skin biopsy treated with aqueous extracts was quantified.

[0016] [ Figure 4 Immunofluorescence assay was performed using 1% red clover (…). Trifolium pratense The expression of neuroprotectin was quantified in human skin biopsy treated with aqueous extracts.

[0017] [ Figure 5 Immunofluorescence assay was performed using 1% quinoa ( Chenopodium quinoa The expression of neuroprotectin was quantified in human skin biopsy treated with aqueous extracts.

[0018] [ Figure 6 Immunofluorescence assay was performed using 1% iris extract (…). Iris pallidaThe expression of neuroprotectin was quantified in human skin biopsy treated with aqueous extracts. Summary of the Invention

[0019] In one embodiment, the present invention relates to peptides having the following general formula (I) that can increase the expression of mitochondrial-derived peptides: R1-(AA)n- Cys-Leu-Leu-Leu-Ser-Thr-X1-(AA)p-R2 in AA represents any amino acid, and n and p are integers between 0 and 2. X1 indicates glycine or no amino acid. R1 represents the primary amine functional group of the N-terminal amino acid, which is either without a protecting group or substituted with a protecting group, wherein the protecting group may be selected from acetyl, benzoyl, toluenesulfonyl, or benzyloxycarbonyl. R1 can also represent palmitoylation, and more particularly S-palmitoylation. R2 represents the hydroxyl functional group of the carboxylic acid of the C-terminal amino acid, which is either without a protecting group or substituted with a protecting group. The protecting group may be selected from an alkyl chain from C1 to C20, or an NH2, NHY, or NYY group, where Y represents an alkyl chain from C1 to C4.

[0020] In another embodiment, the present invention relates to a cosmetic composition or pharmaceutical composition comprising, in a physiologically acceptable medium, an effective amount of an active ingredient or any mixture thereof capable of increasing the expression of mitochondrial-derived peptides, said active ingredient being selected from: the peptide of SEQ ID No. 1, the peptide of SEQ ID No. 2, date palm (… Phoenix dactylifera Aqueous extracts of fruit pulp, germinated red clover ( Trifolium pratense Aqueous extracts of quinoa seeds, quinoa ( Chenopodium quinoa Aqueous extracts of seeds and iris root ( Iris pallida Aqueous extracts of the root, or any mixture thereof.

[0021] In another embodiment, the present invention relates to a cosmetic treatment method aimed at protecting the skin and appendages from external aggressors and combating skin aging, comprising applying a composition according to the invention to the skin. Detailed Description of the Invention definition

[0022] Unless otherwise stated, all terms used in this specification have the broadest known meanings. For the purposes of this invention, the following terms are defined as follows:

[0023] When describing a range of values, the limitation of the range should be understood to explicitly include all intermediate values ​​within the range. For example, a range of values ​​between 1% and 10% should be understood to include 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, as well as all decimal values ​​between 1% and 10%.

[0024] Unless otherwise stated, numerical percentage values ​​are weight percentages, that is, the weight of the compound relative to the total weight of the intended mixture.

[0025] The compositions described in this application may “comprising,” “composed of,” or “substantially consist of” essential compounds or optional ingredients.

[0026] "consistently made of" means that a composition or component may include additional ingredients, but only if the additional ingredients do not alter the essential or novel characteristics of the composition or use described in this application.

[0027] "Physiologically acceptable media" refers to a mediator that is suitable for contact with the outer layer of skin or mucous membranes without toxicity, irritation, excessive allergic reactions or intolerance, and is proportionate to a reasonable benefit / risk ratio.

[0028] "Topical application" means applying or smearing, according to the invention, a peptide or plant extract obtained from the plant or fungal family of interest, or a composition containing the same, onto the surface of the skin or mucous membrane.

[0029] "Skin" refers to the skin including its appendages (hair and body hair, eyebrows, eyelashes, nails).

[0030] It should be understood that this invention relates to mammals, and more particularly to humans.

[0031] The inventors have identified several peptides and plant extracts containing those peptides capable of regulating mitochondrial-derived peptides. In particular, they have demonstrated that some plant extracts or peptides, when applied to the skin, can significantly stimulate mitochondrial function. This has been demonstrated by increased expression of mitochondrial-derived peptides associated with skin benefits, including neuroprotectin (e.g., increased expression of collagen I).

[0032] As used in this article, “active ingredient” refers to any peptide or plant extract that can activate mitochondrial-derived peptides when applied to the skin of a subject as required.

[0033] "Anti-skin or skin aging" includes, but is not limited to, one or more of the following: (a) treating, reducing and / or preventing fine lines or wrinkles; (b) improving skin thickness, plumpness and / or firmness; (c) improving skin suppleness and / or softness; (d) improving procollagen and / or collagen production; (e) improving skin texture and / or promoting densification; (f) improving skin barrier repair and / or function; (g) treating and / or preventing skin sagging or atrophy; (h) improving the appearance of skin contour; (i) restoring skin radiance and / or brightness; (j) improving the appearance of skin reduced due to menopause; (k) improving skin hydration and / or moisture retention; and (l) improving skin elasticity and / or resilience.

[0034] "Active ingredients that can activate mitochondrial-derived peptides" are understood to be any substance that can increase the expression of mitochondrial-derived peptides by activating protein synthesis (through direct or indirect regulation of gene expression), or by increasing the biological activity of mitochondrial-derived peptides, or by other biological processes such as the stabilization of mitochondrial-derived peptides or even the stabilization of messenger RNA transcripts.

[0035] Preferably, the substance capable of activating the mitochondrial-derived peptides according to the present invention is a plant extract or a pure peptide.

[0036] This disclosure covers peptides designed by the inventors by selecting short amino acid sequences of neuroprotective agents (including sequences involved in the neuroprotective activity of neuroprotective agents and those binding to proteins Bax and IGFBP-3). Several peptide sequences were designed to potentially act on neuroprotective agent mitochondrial-derived peptides, but only peptides showing homology with plant sequences were selected.

[0037] Therefore, the first object of the present invention is a peptide capable of activating mitochondrial-derived peptides of general formula (I). R1-(AA)n- Cys-Leu-Leu-Leu-Ser-Thr-X1-(AA)p-R2 in AA represents any amino acid, and n and p are integers between 0 and 2.

[0038] In a particular implementation scheme, X1a indicates glycine or no amino acid.

[0039] R1 represents the primary amine functional group of the N-terminal amino acid, which is either without a protecting group or substituted with a protecting group, wherein the protecting group may be selected from acetyl, benzoyl, toluenesulfonyl, or benzyloxycarbonyl. R1 may also represent palmitoylation, and more particularly S-palmitoylation.

[0040] R2 represents the hydroxyl functional group of the carboxylic acid of the C-terminal amino acid, which is either without a protecting group or substituted with a protecting group. The protecting group may be selected from an alkyl chain from C1 to C20, or an NH2, NHY, or NYY group, where Y represents an alkyl chain from C1 to C4.

[0041] According to a specific embodiment of the present invention, the bioactive peptide consists of the following sequence: (SEQ ID No. 1) Ser-Cys-Leu-Leu-Leu-Ser-Thr,

[0042] According to another specific embodiment of the present invention, the bioactive peptide consists of the following sequence:

[0043] (SEQ ID No. 2) Ser-Cys-Leu-Leu-Leu-Ser-Thr-Gly

[0044] This invention also relates to homologous forms of these sequences. According to the invention, the term "homologous" means any peptide sequence that is at least 50%, or preferably at least 80%, and even more preferably at least 90% identical to the peptide sequences selected from sequences SEQ ID No. 1 to SEQ ID No. 2. "At least X% identical to a peptide sequence" is understood to represent the percentage of identity between the amino acid residues of the two sequences obtained after optimal alignment of the two sequences to be compared. Optimal alignment is obtained using algorithms for local homology, such as those used by the computer software BLAST P or T BLAST N available on the National Center for Biotechnology Information (NCBI) website.

[0045] The term "homologous" can also refer to peptides with sequences different from those of peptides having sequences SEQ ID No. 1 to SEQ ID No. 2, differing only in the substitution of chemically equivalent amino acids, i.e., one residue being replaced by another residue with the same characteristics. Therefore, classical substitutions can occur between Ala, Val, Leu, and Ile; between Ser and Thr; between acidic residues of Asp and Glu; between Asn and Gln; and between basic residues of Lys and Arg, or between aromatic residues of Phe and Tyr.

[0046] The term "peptide" refers to a chain of two or more amino acids linked together by peptide bonds or modified peptide bonds.

[0047] A “peptide” must be understood as at least one fragment of a synthetic peptide or protein, obtained by the proteolysis of a larger protein or polypeptide.

[0048] The peptides of general formula (I) according to the present invention can be obtained from constituent amino acids or derivatives thereof by classical chemical synthesis (in a solid or homogeneous liquid phase) or by enzymatic synthesis (Kullman et al. (1980), J. Biol. Chem., 225, 8234).

[0049] Peptides having general formula (I) can also be obtained by fermentation of genetically modified or unmodified bacterial strains, or even by extracting proteins of animal or plant origin, preferably of plant origin, followed by controlled hydrolysis that releases peptide fragments that correspond wholly or partially to peptides having general formula (I).

[0050] Those skilled in the art of protein and peptide synthesis, extraction, and purification can conceive of other procedures, whether simpler or more complex. Therefore, the peptides according to the invention can be of natural or synthetic origin. Preferably, according to the invention, the peptides of general formula (I) are obtained through chemical synthesis.

[0051] Many proteins found in plants may contain peptides of general formula (I) within their structures. Controlled hydrolysis allows these peptide fragments to be released. To realize the present invention, it may be possible, but not necessary, to first extract the protein of interest and then hydrolyze it, or to first hydrolyze the original extract and then purify the peptide fragments. It is also possible to use certain hydrolyzed extracts without purifying the peptide fragments corresponding to general formula (I) according to the invention, while simultaneously ensuring the presence of said fragments through appropriate analytical methods.

[0052] In the second step, the inventors selected plants that have sequence homology with the peptide of general formula (I) in order to provide plant extracts that have the same biological activity as the peptide of general formula (I).

[0053] Sequence similarity searches were performed using comprehensive protein sequence databases such as BLAST according to standard procedures (William Pearson, “An Introduction to Sequence Similarity (“Homology”) searching”, Curr Protoc Bioinformatics. June 2013). Homologous sequences were identified.

[0054] This search allows inventors to identify several candidates in the following classification groups: o Arecaceae ( Arecaceae ), genus *Solanum* Phoenix ), especially dates ( Phoenix dactylifera )kind. o Leguminosae ( Fabaceae ), Clover ( Trifolium ), especially clover (Trifolium pratense )kind. o Iris family ( Iridaceae ), Iris genus ( Iris ), especially iris root ( Iris pallida )kind. o Amaranthaceae ( Amaranthaceae ), genus Chenopodiaceae ( Chenopodium ), especially quinoa ( Chenopodium quinoa )kind. o Anacardiaceae ( Anacardiaceae ), Pistacia genus ( Pistacia ), especially A-Yuehunzi ( Pistacia vera ) and Atlantic Pistacia chinensis ( Pistacia atlantica )kind. o Sapindaceae ( Sapindaceae ), Xanthoceras genus ( Xanthoceras ), especially the Chinese tallow tree ( Xanthoceras sorbifolia )kind. o Anomala ( Teratosphaeriaceae ), Friedmanella spp. Friedmanniomyces Ascomycota ( Ascomycota ), especially endophytic fungal species, such as Friedmann's endophytes ( Friedmanniomyces endolithicus ). o Marine Spirogyra ( Oceanospirillaceae Marine Monoclonal Bacteria ( Marinomonas ), especially species of the genus *Marine Monotrophus*, such as *Marine Monotrophus thuringiensis* (Mediterranean Marine Monotrophus). Marinomonas mediterranea ).

[0055] A series of peptides different from those having the general formula (I) were identified as potential agents for neuroprotectin. However, these peptides were not selected because they did not show homology with plant sequences.

[0056] In one specific embodiment, the present invention relates to an aqueous extract of date fruit pulp obtained by crushing the pulp in water and subjecting the plant material to microwave, having a dry matter content of 20-50 g / kg.

[0057] In another specific embodiment, the present invention relates to germinating red clover ( Trifolium pratense An aqueous extract of the seeds, having a dry matter content of 15 to 30 g / kg, obtained using the method described in patent S11,021,505 B2 (incorporated herein by reference).

[0058] In another specific embodiment, the present invention relates to quinoa obtained by crushing seeds in water and subjecting the plant material to microwaves. Chenopodium quinoaAqueous extracts of the seeds, containing 4 to 10 g / kg of dry matter.

[0059] In another particular embodiment, the present invention relates to an aqueous extract of iris root having a dry matter content of 8 g / kg to 14 g / kg and obtained using the method described in patent US11,021,505 B2 (incorporated herein by reference).

[0060] According to the present invention, the active ingredient may be a pure peptide, a mixture of several peptides, or at least one aqueous plant extract containing a peptide. The effective amount of the active ingredient corresponds to the minimum amount required to obtain the desired result (i.e., increased expression of the mitochondrial-derived peptide neuroprotectin in skin cells).

[0061] A second object of this disclosure is a cosmetic or pharmaceutical composition comprising, in a physiologically acceptable medium, an effective amount of at least one of the aforementioned peptides or an aqueous extract or any mixture thereof as an active ingredient, capable of activating mitochondrial-derived peptides.

[0062] In one advantageous embodiment, the composition is a cosmetic composition.

[0063] In one specific embodiment, the cosmetic composition contains, in a physiologically acceptable medium, an effective amount of an active ingredient capable of increasing the expression of mitochondrial-derived peptides, said active ingredient being selected from: the peptide of SEQ ID No. 1, the peptide of SEQ ID No. 2, and date palm (…). Phoenix dactylifera) Aqueous extracts of fruit pulp, sprouted red clover ( Trifolium pratense Aqueous extracts of quinoa seeds, quinoa ( Chenopodium quinoa Aqueous extracts of seeds and iris root ( Iris pallida Aqueous extracts of the root or any mixture thereof.

[0064] Preferably, the mitochondrial-derived peptide is a neuroprotective agent.

[0065] According to an advantageous embodiment, when the active ingredient is a peptide of SEQ ID No. 1 or SEQ ID No. 1, its concentration is about 0.0005-500 mg / kg (parts per million) relative to the total weight of the final composition, preferably about 0.01-10 mg / kg.

[0066] According to another embodiment, when the active ingredient is an aqueous extract, its concentration is preferably from about 0.01% to 5% relative to the total weight of the final composition, and more preferably from about 0.1% to 2%.

[0067] According to another embodiment, the active ingredient according to the invention is pre-diluted in one or more physiologically acceptable solvents, such as water, glycerin, ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diols, cyclic polyols, petrolatum, vegetable oils, or any mixture of these solvents.

[0068] According to a specific implementation scheme, when the active ingredient is an aqueous extract of iris root, it can be diluted in 20% propylene glycol (propylene-1,3-diol) and 10% glycerin.

[0069] Of course, it should be understood that the active ingredients according to the invention can be used alone or in combination with at least one other active ingredient for use in cosmetic compositions or for the preparation of pharmaceutical and / or dermatological compositions.

[0070] The compositions according to the invention can be applied in any suitable manner, particularly orally, parenterally or topically, and their formulation will be adjusted by a professional, particularly for cosmetic or dermatological compositions.

[0071] Advantageously, the composition according to the invention is in a form suitable for application by local means.

[0072] The compositions available according to the invention may be in particular in the form of creams, oil-in-water emulsions or water-in-oil or multiple emulsions, solutions, suspensions, gels, lotions, lotions, sticks or even powders, suitable for application to the skin, lips and / or appendages.

[0073] The compositions may include excipients required for their formulation, such as solvents, thickeners, diluents, surfactants, antioxidants, colorants, preservatives, or fragrances.

[0074] Advantageously, the available compositions contain at least one other active agent that promotes the action of the peptide according to the invention. Therefore, the compositions according to the invention can be associated with the active ingredient according to the invention, which has antioxidant activity, or stimulates the synthesis of dermal macromolecules, or stimulates energy metabolism. For example, vitamin C, vitamin E, coenzyme Q10, and polyphenolic extracts from plants can be cited as active agents with anti-free radical or antioxidant activity.

[0075] The compositions may be, in particular, aqueous, hydroalcoholic, or oily solutions, oil-in-water or water-in-oil emulsions, or a combination of these emulsions. They may also be in the form of creams, suspensions, or even powders suitable for application to the skin, mucous membranes, lips, and / or appendages. These compositions may be more or less fluid and have the appearance of cream, lotion, milk, butter, ointment, gel, paste, or mousse. They may also be in solid form, such as sticks, or applied to the skin as aerosols. They can be used as care products and / or as cosmetics for the skin.

[0076] In addition, the composition may include any additives commonly used in the intended application area, as well as adjuvants required for their formulation, such as solvents, thickeners, diluents, antioxidants, colorants, solar filters, self-tanning agents, pigments, carriers, preservatives, fragrances, deodorants, active cosmetic or pharmaceutical ingredients, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0077] In any case, those skilled in the art will note that these adjuvants and their proportions are selected in a manner that does not impair the advantageous properties studied in the compositions according to the invention. These adjuvants may, for example, be equivalent to 0.01 to 20% of the total weight of the composition. When the compositions of the invention are emulsions, the fatty phase may comprise 5 to 80% by weight, preferably 5 to 50% by weight, relative to the total weight of the composition. The emulsifiers and co-emulsifiers used in the compositions will be selected from those conventionally used in the fields under consideration. For example, they may be used at a weight ratio of 0.3 to 30% relative to the total weight of the composition.

[0078] In another embodiment, the present invention relates to a cosmetic treatment method aimed at protecting the skin and appendages from external damage and combating skin aging, comprising applying a composition according to the invention to the skin of a subject in need.

[0079] In another embodiment, the present invention relates to a cosmetic treatment method aimed at increasing the expression of mitochondrial-derived peptides and collagen I in skin cells.

[0080] Advantageously, mitochondrial-derived peptides are brain protectants. Example

[0081] As an illustration, an exemplary embodiment of the process according to the present invention is described below.

[0082] Example 1: Effects of peptides from SEQ ID No. 1 and SEQ ID No. 2 on the expression of neuroprotectin in isolated human skin biopsy tissue

[0083] The aim of this study was to determine the effects of the peptides in SEQ ID No. 1 and SEQ ID No. 2 on the expression of human neuroprotectin mitochondrial-derived peptides.

[0084] Protocol: Normal human skin biopsy tissue was treated twice daily for 2 days with a 1% diluted solution of 500 mg / kg SEQ ID No. 1 or SEQ ID No. 2 in PBS. PBS was used as a negative control. For immunolabeling with anti-cerebroside antibody, the tissue was fixed and embedded in paraffin. The embedded skin biopsy tissue was then cut, and the sections were dewaxed and rehydrated. A demasking protocol was then performed, followed by administration of a specific anti-human cerebroside antibody (Sigma, ref. H2414, rabbit polyclonal), and then administration of a second suitable antibody conjugated to a fluorescent dye. After mounting in a specific medium, the slides were observed using an epifluorescence microscope (Nikon Eclipse 80i microscope).

[0085] Results: Biopsy tissue compared with PBS control ( Figure 1 Compared to treatment with 5 mg / Kg of SEQ ID No.1 and SEQ ID No.2 peptides, microscopic observation of skin sections after immunofluorescence staining of cerebrolysin showed increased expression of cerebrolysin.

[0086] Example 2: Effects of peptides from SEQ ID No. 1 and SEQ ID No. 2 on collagen I expression in isolated human skin biopsy tissue

[0087] The aim of this study was to determine the effect of the peptides in SEQ ID No. 1 and SEQ ID No. 2 on the expression of collagen I.

[0088] Protocol: Normal human skin biopsies were treated twice daily for 2 days with a 1% diluted solution of 500 mg / kg SEQ ID No. 1 or SEQ ID No. 2 in PBS. PBS was used as a negative control. For immunolabeling with anti-collagen I antibody, the tissue was fixed and embedded in paraffin. The embedded skin biopsies were then cut, and the sections were dewaxed and rehydrated. A demasking protocol was then performed, followed by administration of a specific anti-collagen I antibody (Proteintech, 14695-1-AP, rabbit polyclonal), and then administration of a second suitable antibody conjugated to a fluorescent dye. After mounting in a specific medium, the slides were observed using an epifluorescence microscope (Nikon Eclipse 80i microscope).

[0089] Results: Compared with PBS control biopsy tissue, microscopic observation of collagen I immunofluorescence staining on skin sections showed increased collagen I expression after treatment with 5 mg / Kg of SEQ ID No. 1 and SEQ ID No. 2 peptides. Figure 2 ).

[0090] Example 3: Effect of date palm extract on the expression of neuroprotectin in isolated human skin biopsy tissue

[0091] The aim of this study was to determine the effect of aqueous extracts of date palms on the expression of neuroprotectin.

[0092] Protocol: Normal human skin biopsy tissue was treated twice daily for 2 days with the date palm extract of Example 7 diluted 1% in PBS. PBS was used as a negative control. For immunolabeling with anti-cerebroside antibody, the tissue was fixed and embedded in paraffin. The embedded skin biopsy tissue was then cut, and the sections were dewaxed and rehydrated. A demasking protocol was then performed, followed by the application of a specific anti-cerebroside antibody (ProteinTech, ref. 25885-1-AP, rabbit polyclonal), and then a second suitable antibody conjugated to a fluorescent dye. After mounting in a specific medium, the slides were observed using an epifluorescence microscope (Nikon Eclipse 80i microscope).

[0093] Results: Biopsy tissue compared with PBS control ( Figure 3 In contrast, microscopic observation of skin sections treated with 1% date extract and stained with immunofluorescence showed increased expression of neuroprotectin.

[0094] Example 4: Effect of red clover extract on the expression of neuroprotectin in isolated human skin biopsy tissue

[0095] The aim of this study was to determine the effect of aqueous extracts of red clover on the expression of neuroprotectin.

[0096] Protocol: Normal human skin biopsy tissue was treated twice daily for 2 days with red clover extract of Example 7 diluted 1% in PBS. PBS was used as a negative control. For immunolabeling with anti-cerebroside antibody, the tissue was fixed and embedded in paraffin. The embedded skin biopsy tissue was then cut, and the sections were dewaxed and rehydrated. A demasking protocol was then performed, followed by administration of a specific anti-cerebroside antibody (Sigma, ref. H2414, rabbit polyclonal), and then administration of a second suitable antibody conjugated to a fluorescent dye. After mounting in a specific medium, the slides were observed using an epifluorescence microscope (Nikon Eclipse 80i microscope).

[0097] Results: Compared with PBS control biopsy tissue, microscopic observation of skin sections treated with 1% red clover extract showed increased expression of cerebrolysin immunofluorescence staining. Figure 4 ).

[0098] Example 5: Effect of quinoa extract on the expression of neuroprotectin in isolated human skin biopsy tissue

[0099] The aim of this study was to determine the effect of the aqueous extract of quinoa from Example 7 on the expression of neuroprotectin.

[0100] Protocol: Normal human skin biopsy tissue was treated twice daily for 2 days with quinoa extract of Example 7 diluted 1% in PBS. PBS was used as a negative control. For immunolabeling with anti-cerebroside antibody, the tissue was fixed and embedded in paraffin. The embedded skin biopsy tissue was then cut, and the sections were dewaxed and rehydrated. A demasking protocol was then performed, followed by the application of a specific anti-human antibody (ProteinTech, ref. 25885-1-AP, rabbit polyclonal), and then a second suitable antibody conjugated to a fluorescent dye. After mounting in a specific medium, the slides were observed using an epifluorescence microscope (Nikon Eclipse 80i microscope).

[0101] Results: Microscopic observation of immunofluorescence staining of skin sections with cinnamicin showed increased expression of cinnamicin after treatment with 1% quinoa extract compared to PBS control biopsy tissue. Figure 5 ).

[0102] Example 6: Effect of Iris tectorum extract on the expression of neuroprotectin in isolated human skin biopsy tissue

[0103] The aim of this study was to determine the effect of an aqueous extract of Iris tectorum root on the expression of neuroprotective peptide mitochondrial-derived peptide.

[0104] Protocol: Normal human skin biopsy tissue was treated twice daily for 2 days with the iris root extract (diluted 1% in PBS) from Example 7. PBS was used as a negative control. For immunolabeling with anti-cerebroside antibody, the tissue was fixed and embedded in paraffin. The embedded skin biopsy tissue was then cut, and the sections were dewaxed and rehydrated. A demasking protocol was then performed, followed by the application of a specific anti-cerebroside antibody (ProteinTech, ref. 25885-1-AP, rabbit polyclonal), and then a second suitable antibody conjugated to a fluorescent dye. After mounting in a specific medium, the slides were observed using an epifluorescence microscope (Nikon Eclipse 80i microscope).

[0105] Results: Compared with PBS control biopsy tissue, microscopic observation of skin sections treated with 1% Iris tectorum root extract showed increased expression of cerebrolysin immunofluorescence staining. Figure 6 ).

[0106] Example 7: Preparation of Aqueous Plant Extracts

[0107] Date Palms: Date palm pulp was crushed, and then 5% of the crushed plant material in water was extracted for 1 to 10 minutes using a high-performance microwave (maximum magnetron 850 watts, maximum pressure 30 bar) at a temperature between 60-80°C. Solids were removed by centrifugation, and the supernatant was collected and clarified by sequential filtration until sterile. The obtained date palm fruit extract had a dry matter content of 20-50 g / kg and was used for bioefficacy testing.

[0108] Red clover: Red clover seeds were germinated by soaking in water. 10% of the germinated seeds were crushed and water extracted using the proprietary technique disclosed in patent US 11,021,505 B2. Solids were then removed by centrifugation, and the supernatant was collected and clarified by sequential filtration until sterile. The resulting red clover extract had a dry matter content between 15-30 g / kg and was used as is for bioefficacy testing.

[0109] Quinoa: Quinoa seeds were crushed and then 5% of the crushed plant was extracted from water using a high-performance microwave (maximum power magnetron 850 watts, maximum pressure 30 bar) at 60-80°C for 1-10 minutes. Solids were then removed by centrifugation, and the supernatant was collected and clarified by sequential filtration until it passed through sterile pores. The obtained quinoa seed extract had a dry matter content of 4-10 g / kg and was used for bioefficacy testing.

[0110] Orris root: Extracting crushed and dried orris root using the proprietary technology disclosed in patent US 11,021,505 B2 by a method comprising the following steps: a) Extracting pulverized, dried roots of *Iris tectorum* root at a concentration of 2.5% in water in the presence of 10 mM phytic acid, at a pH of 8 to 11, preferably 10.5 to 11, at a temperature of 40 to 80°C, for a duration of 30 minutes to 4 hours, preferably 2 hours. b) Remove solids by centrifugation and collect the supernatant. c) Clarify the supernatant by sequential filtration until sterilization is achieved. d) The filtered extract was then diluted in water to obtain an orris root extract with a dry matter content of 8 to 14 g / kg. The diluted extract was used as is for bioefficacy testing. In step d), the iris root extract may alternatively be diluted in a solvent, such as water, glycerin, ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diethylene glycol, cyclic polyols, petrolatum, vegetable oils, or any mixture of these solvents. Preferably, the iris root extract is diluted in 20% propylene glycol (propylene-1,3-diol) and 10% glycerin. sequence list

Claims

1. A peptide capable of increasing the expression of mitochondrial-derived peptides, having the following general formula (I): R1-(AA)n- Cys-Leu-Leu-Leu-Ser-Thr-X1-(AA)p-R2 in, AA represents any amino acid, and n and p are integers between 0 and 2. X1 indicates glycine or no amino acid. R1 represents the primary amine functional group of the N-terminal amino acid, which is either without a protecting group or substituted with a protecting group. The protecting group can be selected from acetyl, benzoyl, toluenesulfonyl, or benzyloxycarbonyl. R1 can also represent palmitoylation, and more particularly S-palmitoylation. R2 represents the hydroxyl functional group of the carboxylic acid of the C-terminal amino acid, which is either without a protecting group or substituted with a protecting group. The protecting group may be selected from an alkyl chain from C1 to C20, or an NH2, NHY, or NYY group, where Y represents an alkyl chain from C1 to C4.

2. The peptide according to claim 1, wherein the amino acid sequence SEQ ID No 1: Ser-Cys-Leu-Leu-Leu-Ser-Thr is composed of the following amino acid sequence: Ser-Cys-Leu-Leu-Leu-Ser-Thr.

3. The peptide according to claim 1, wherein it comprises the amino acid sequence SEQ ID No 2: Ser-Cys-Leu-Leu-Leu-Ser-Thr-Gly.

4. A cosmetic or pharmaceutical composition comprising, in a physiologically acceptable medium, an effective amount of an active ingredient capable of increasing the expression of mitochondrial-derived peptides, said active ingredient being selected from: the peptide of SEQ ID No. 1, the peptide of SEQ ID No. 2, and date palm (…). Phoenix dactylifera Aqueous extracts of fruit pulp, germinated red clover ( Trifolium pratense Aqueous extracts of quinoa seeds, quinoa ( Chenopodium quinoa Aqueous extracts of seeds and iris root ( Iris pallida Aqueous extracts of the root, or any mixture thereof, or any mixture thereof.

5. The composition according to claim 4, wherein the mitochondrial-derived peptide is a neuroprotective agent.

6. The composition according to claim 4, wherein when the active ingredient is a peptide of SEQ ID No: 1 or SEQ ID No: 2, the concentration of the active ingredient is between about 0.0005 mg / kg and 500 mg / kg, and preferably between about 0.01 mg / kg and 10 mg / kg, relative to the total weight of the final composition.

7. The composition according to claim 4, wherein when the active ingredient is an aqueous extract, the concentration of the active ingredient relative to the total weight of the final composition is between about 0.01% and 5%, preferably between about 0.1% and 2%.

8. The composition according to claim 4, wherein the aqueous extract of the iris root is obtained by a method comprising the following steps: a) extracting pulverized iris root in water at a concentration of 2.5% in the presence of 10 mM phytic acid, at a pH of 8 to 11, preferably 10.5 to 11, at a temperature of 40 to 80°C for a duration of 30 minutes to 4 hours, preferably 2 hours; b) removing solids by centrifugation and collecting the supernatant; c) clarifying the supernatant by sequential filtration until sterilization is achieved; and d) diluting the filtered supernatant in water to obtain an extract with a dry matter content of 8 to 14 g / kg.

9. The composition according to claim 8, wherein in step d), the aqueous extract of the iris root is diluted in a solvent selected from the group consisting of water, glycerin, ethanol, propylene glycol, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diethylene glycol, cyclic polyols, petrolatum, vegetable oils, and any mixture of these solvents.

10. The composition according to claim 8, wherein in step d), the iris root extract is diluted in 20% propylene glycol (propylene-1,3-diol) and 10% glycerol.

11. The composition according to claim 4, wherein the composition is in the form of a cream, an oil-in-water emulsion or a water-in-oil or multiple emulsion, a solution, a suspension, a gel, an emulsion, a lotion, a stick or even a powder.

12. The composition of claim 4, wherein the composition is suitable for topical application to the skin, lips and / or appendages.

13. A cosmetic treatment method designed to protect skin and appendages from external aggressors and to combat skin aging, the method comprising applying to the skin the composition according to claim 4.

14. The cosmetic treatment method according to claim 13, wherein the applied composition increases the expression of neuroprotectin in skin cells.

15. The cosmetic treatment method according to claim 13, wherein the applied composition increases the expression of collagen I in skin cells.

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