Method for evaluating the ability of a substance or composition to prevent, slow down or eliminate the signs of aging of the skin or lips

Abstract

The present invention relates to a method for evaluating the ability of a chemical substance (S) or a chemical composition (C) to prevent or slow down the appearance of signs of aging on human skin or lips, or even to eliminate said signs; said method comprising: - step a) contacting the chemical substance or the chemical composition with "young" human dermal fibroblasts at passage R6 obtained from a culture medium; - step b) contacting senescent cells of the human dermal fibroblasts with "young" fibroblasts from step a); and - step c) measuring the senescence input of the "young" fibroblasts and comparing it with a reference value.

Description

[0001] The present invention relates to a method for evaluating the ability of a chemical substance (S) or a chemical composition (C) to prevent or slow down the appearance of or eliminate the signs of aging of human skin or lips.

[0002] The invention also relates to cosmetic compositions and cosmetic formulations.

[0003] The human skin constitutes the first visual image presented to others, and therefore, improving its appearance is generally a subject of human concern. The skin is a reflection of a state of health, which is usually associated with youth, or, on the contrary, of a state of fatigue and / or aging. Therefore, cosmetic consumers are looking for solutions for reducing and / or preventing the visible manifestations associated with said aging.

[0004] Skin aging is a phenomenon caused by two different biological processes:

[0005] - Intrinsic or "chronological" aging that is dependent on genetic traits and is therefore specific to each individual

[0006] - Extrinsic aging influenced by environmental factors, such as repeated and prolonged exposure to natural or artificial ultraviolet light (or photoaging), atmospheric pollution, cigarette smoke, various oxidants; various psychological, emotional and / or neurological stresses.

[0007] Aging is characterized by metabolic, functional, cellular, architectural and tissue impairments that lead to visible external effects. For skin aging, key issues are the appearance and / or growth of wrinkles and fine lines, dull skin tone, lack of uniformity of skin tone (skin discoloration), or changes in skin texture (e.g. reduced skin thickness) and in the properties of the skin, in particular its biomechanical properties (loss of elasticity).

[0008] Thus, over a period of time, the renewal of the skin (which is an organ in constant renewal) slows down, a phenomenon which leads to the accumulation of "old" cells and to the impairment of their repair and maintenance mechanisms.

[0009] Skin cells then enter a state of senescence: they undergo a cell cycle arrest following a limited number of divisions, which is associated with numerous changes and specific phenotypic traits [1].

[0010] The most notable changes are:

[0011] - Increased cell size

[0012] - Increased nuclear size

[0013] - Increased lysosomal activity leading to increased autophagy

[0014] - Production and secretion of bioactive molecules, such as interleukins, growth factors, matrix-degrading enzymes or reactive oxygen species (ROS); these bioactive molecules constitute the senescence-associated secretory phenotype (SASP) or the senescence-message secretome (SMS)[2][3]

[0015] - Changes in metabolic activity were significantly associated with an increase in the synthesis of matrix metalloproteinase (MMP-1), a decrease in the synthesis of procollagen-1, and an increase in the activity of β-galactosidase, a characteristic marker of cellular senescence

[0016] - DNA damage

[0017] -Alterations in the cell cycle and slowing of cell proliferation.

[0018] This aging phenomenon is associated with multiple mechanisms, including the infection of "young" cells with mediators produced by neighboring senescent cells.

[0019] These molecules (SMS or SASP) secreted by "old" cells are detected by neighboring "young" cells that do not show the above-mentioned cellular changes (morphological, metabolic or proliferation, etc.). These neighboring "young" cells that have come into contact with the molecules (or SMS) secreted by senescent cells will then proceed to enter a senescent state and show the specific morphology and cellular damage of skin aging. This phenomenon has been recently described and has been called "contagious aging" or the "bystander effect" [4].

[0020] Methods using co-cultures of different skin cells exposed to UV light have been described for studying the sensitivity of human melanocytes, keratinocytes, and fibroblasts to induce or be susceptible to the "bystander effect" [5]. In these methods, different cell cultures were exposed to UVA rays and various markers of aging, particularly oxidation and DNA damage, were assessed. From this research:

[0021] - Aging can be slightly induced by direct exposure of melanocytes to UV-A rays, but the melanocytes are susceptible to infectious aging or the "bystander effect";

[0022] - Keratinocytes are the cells most resistant to infectious senescence or the "bystander effect";

[0023] -Fibroblasts showed a moderate response to both induction by ultraviolet A rays and resistance to infectious senescence or the "bystander effect."

[0024] However, this method is characterized by a long implementation time and is not very suitable for use in screening methods for selecting anti-aging active agents, which is very common in the research field.

[0025] It does not appear that the prior art discloses any in vitro method for selecting chemical substances or chemical compositions for preventing or slowing down the appearance of or eliminating the signs of aging on human skin or lips, which in vitro method is easy to use in screening methods with medium throughput, does not involve animal cells, is reproducible, sensitive, discriminating and effective, and which in vitro method is suitable for the cosmetic market.

[0026] Therefore, there is a need to develop methods for selecting chemicals or chemical compositions that prevent or slow the appearance of or eliminate signs of aging on human skin or lips; said methods:

[0027] - Does not involve the use of animal cells

[0028] - Conducted within a sufficiently short period of time (maximum 72 hours)

[0029] - Not using any clinical research models or models derived from biopsies from clinical studies

[0030] - and can therefore be used in a screening method aimed at testing and possibly selecting up to 12 products distributed in four culture series.

[0031] One solution of the present invention is a method for evaluating the ability of a chemical substance (S) or a chemical composition (C) to prevent or slow down the appearance of signs of aging on human skin or lips or to eliminate said signs; said method comprising:

[0032] - step a) contacting a chemical substance or a chemical composition with "young" human dermal fibroblasts at passage R6 obtained from culture,

[0033] - step b) contacting human aged dermal fibroblasts with "young" fibroblasts obtained from step a), and

[0034] - Step c) measurement of the entry of "young" fibroblasts into senescence and comparison with reference values.

[0035] The term "substance" means a molecule defined by a formula, and the term "composition" means a mixture of molecules.

[0036] The term "contacting" means adding in the same medium.

[0037] For the purposes of the present invention, the term "human dermal fibroblasts" refers to cells present in the connective tissue, located in the dermis and ensuring its coherence and softness. A person skilled in the art will particularly appreciate that during skin aging, the number of fibroblasts has a tendency to decrease, the ratio between collagen type III fibers and collagen type I fibers changes, and the elements of the extracellular matrix (collagen, elastic fibers, glycosaminoglycans) decrease. The skin becomes thinner, loses elasticity, and develops folds or wrinkles.

[0038] For the purposes of the present invention, the term "Ri passage" refers to the number of subcultures (i) that a cell undergoes, i.e.:

[0039] - R19 passage means 19 subcultures

[0040] -R20 passage means 20 subcultures

[0041] - R5 passage means 5 subcultures

[0042] - R6 passage means 6 subcultures.

[0043] For the purposes of the present invention, the term "ability to prevent or slow down the appearance of or eliminate the signs of aging on human skin or lips" means the ability to prolong the duration before the appearance of said signs of aging on human skin or lips.

[0044] For the purposes of the present invention, the term "signs of aging of human skin or lips" is intended to mean any change in the appearance of the skin or lips due to aging, such as wrinkles and fine lines, impaired microtopography of the human skin or lips, lack of elasticity and / or tone, lack of density and / or firmness of the skin, but also any internal changes in the skin that are not organically reflected by an altered appearance (such as any internal degradation of the skin due to continuous exposure to UV radiation).

[0045] Depending on the circumstances, the method according to the invention may have one or more of the following features:

[0046] - Senescent cells are human dermal fibroblasts at passage R20 obtained from culture medium,

[0047] - Senescent cells are human dermal fibroblasts at passage R19 that have been harvested from culture before undergoing expansion,

[0048] - "young" human dermal fibroblasts at passage R6 obtained from culture medium are obtained by expansion of "young" human dermal fibroblasts at passage R5 obtained from culture medium,

[0049] - step c) of measuring the entry of "young" fibroblasts into senescence comprises the use of at least two biomarkers: at least one biomarker for SASP / SMS (M1) and at least one biomarker for senescence (M2),

[0050] - biomarkers of SASP / SMS (M1) are selected from the group consisting of proinflammatory cytokines, and more particularly extracellular interleukin-1 (IL-1), extracellular interleukin-6 (IL-6), extracellular interleukin-8 (IL-8), growth factors, matrix degrading enzymes (such as MMP-1 and MMP-3), reactive oxygen species or ROS,

[0051] - an element of a biomarker of aging (M2) selected from the group consisting of: β-galactosidase, persistent DNA damage and apoptosis resistance,

[0052] - biomarker (M1) is interleukin-6 and biomarker (M2) is beta-galactosidase,

[0053] - in step c), comparing the expression level of the two biomarkers (M1) and (M2) with at least one reference expression level of each of the two biomarkers,

[0054] - Signs of ageing of the human skin or lips selected from the group consisting of the appearance of wrinkles, fine lines or impaired microtopography; and / or the appearance of a lack of elasticity and / or tone; and / or the appearance of a lack of density and / or firmness.

[0055] For the purposes of this patent application, the term "biomarker" means a characteristic that is objectively measured and evaluated as an indicator of a normal biological process, a pathogenic process, or a pharmacological response to an external intervention. A biomarker can be, for example, a substance whose detection indicates a specific pathological state, or conversely, a substance whose detection indicates a specific physiological state.

[0056] For the purposes of the present invention, the term "biomarker of the SASP / SMS (M1)" means a biomarker as previously defined, the changes in the expression of which are associated with infectious aging or bystander effect phenomena (in the sense of belonging to the group of active molecules contained in the SASP or "senescence-associated secretory phenotype" (also called SMS or "senescence message secretome")).

[0057] Among the biomarkers (M1) of SASP / SMS, the expression levels of which are measured in step e) of the method that is the subject of the present invention, mention may be made of the elements of the group of biomarkers (M1) consisting of extracellular interleukin-1 (IL-1), extracellular interleukin-6 (IL-6), interleukin-8 (IL-8), growth factors, matrix degrading enzymes (such as MMP-1 and MMP-3), reactive oxygen species or ROS, and in particular extracellular interleukin-6 (IL-6).

[0058] For the purposes of this patent application, the biomarker IL-6 comprises a human IL-6 gene (NCBI reference: gene ID: gene bank: JQ250825.1), and the product of the gene. In a specific embodiment, the biomarker IL-6 consists of one of the products of the human IL-6 gene. The product of the human IL-6 gene comprises a transcript of the human IL-6 gene and a human IL-6 protein. For the purposes of this patent application, a "transcript of the human IL-6 gene" is a polynucleotide whose sequence has a NCBI reference: gene bank: M54894.1. For the purposes of this patent application, the term "human IL-6 protein" means a protein whose peptide sequence is a sequence of the NCBI reference sequence gene bank: AAD13886.1.

[0059] For the purpose of this patent application, the term "biomarker of senescence (M2)" means a biomarker as defined previously, the changes in the expression of which are associated with cellular senescence.

[0060] Among the biomarkers of aging (M2), the expression levels of which are measured in step c) of the method that is the subject of the present invention, mention may be made of the elements of the group of biomarkers (M2) consisting of: β-galactosidase, persistent DNA damage (associated with the detection of γH2AX foci), resistance to apoptosis (associated with the detection of p53, p16, p21 and IGFBP5), most particularly β-galactosidase.

[0061] According to a particular aspect of the method as previously defined, in step c), the biomarker (M1 ) is interleukin-6 and the biomarker (M2) is β-galactosidase.

[0062] For each biomarker (M1) and (M2), expression "measurement of expression level" generally refers to measurement of the amount of gene transcript, or to measurement of the amount of biomolecules, and more particularly proteins and metabolites produced by the human body, or the level of enzyme activity.

[0063] When measuring the expression level of a biomarker at the nucleotide level (i.e. by measuring the amount of a gene product in its nucleotide form), any method commonly used by a person skilled in the art for measuring the amount of nucleotides can be used, and thus qRTPCR (real-time quantitative polymerase chain reaction), DNA chips and in situ hybridization can be mentioned.

[0064] When measuring the expression level of a biomarker at the protein, functional or metabolic level, i.e. by measuring its amount if a protein or metabolite, or by measuring its biological function, any method commonly used by a person skilled in the art for measuring the amount or function of a protein or metabolite can be used, and thus ELISA assays, Western blotting, mass spectrometry, immunofluorescence, chromatography techniques and enzyme activity may be mentioned.

[0065] In the method that is the subject of the present invention, the measurement of the expression of a biomarker of inflammation (M1) is carried out more particularly at the protein level.

[0066] According to a more specific aspect of the method as the subject of the present invention, when the biomarker (M1) is interleukin-6, the measurement of interleukin-6 expression is carried out at the protein level, and even more specifically using a colorimetric ELISA (enzyme-linked immunosorbent assay) method for measurement. The method uses an antibody specific to human IL-6, and the antibody is attached to the bottom of the well of the measurement plate. The sample to be measured is added to the well, and the IL-6 present in the sample is combined with the fixed antibody. The wells are then washed, and another biotinylated anti-human IL-6 antibody is added.

[0067] The wells are washed again to remove unbound antibodies and streptavidin conjugated to the enzyme HRP (horseradish peroxidase) is added to the wells. The wells are washed once more and the HRP substrate tetramethylbenzidine is added to the wells.

[0068] The intensity of the color develops in proportion to the amount of bound IL-6. The "stop" solution changes color from blue to yellow, and the color intensity is measured spectrophotometrically at 450 nm. The IL-6 concentration is calculated by reference to the calibration range.

[0069] The measurement of β-galactosidase activity is performed at the metabolic level, even more particularly by an assay involving colorimetric enzymatic detection.

[0070] Briefly, the β-galactosidase present in the cells reacts specifically with one of its substrates, X-gal (or 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (C14H15BrClNO6)) deposited on the cells. The substrate is a galactoside, a galactose heteroside, bound to a substituted indole nucleus. This colorless compound is hydrolyzed by the β-galactosidase, releasing the indole moiety, which then forms a water-insoluble blue compound that precipitates at the reaction site through oxidation. The blue coloration is detected / observed using a microscope capable of taking pictures. For each picture, the number of labeled cells (blue cells) and the total number of visible cells are counted, and the number of blue cells is taken as a ratio of the total number of cells for normalization.

[0071] According to an even more specific aspect of the method that is the subject of the invention, when the biomarker (M1) is interleukin-6, the measurement of interleukin-6 expression is carried out at the protein level, even more particularly by an assay using a colorimetric ELISA method, and when the biomarker (M2) is β-galactosidase, the measurement of β-galactosidase activity is carried out at the metabolic level, even more particularly by an assay using a colorimetric enzymatic assay.

[0072] The term "reference expression level" of a biomarker refers to any expression level of said biomarker used as a reference.

[0073] In the context of the present invention, the reference expression level of a biomarker (M1) of SASP / SMS can be obtained by measuring the expression level of the biomarker (M1):

[0074] - in a cell culture medium of "young" human dermal fibroblasts at passage R6 (the "young" cells at passage R6 were not brought into contact with senescent cells nor treated with the substance or chemical composition). This measurement of the expression level of the marker (M1) of untreated and unstressed cells is recorded as N1, and / or

[0075] - in a cell culture medium obtained by contacting "young" human dermal fibroblasts obtained at passage R6 with aged human dermal fibroblasts obtained at passage R20; recording this measurement of the expression level of the marker (M1) of untreated but stressed cells as N10; and / or

[0076] - in the cell culture medium obtained at the end of the implementation of step b) of the method that is the subject of the invention, when the substance (S) used in step a) of the method that is the subject of the invention is a substance of an element chosen from the group consisting of vitamin E and vitamin C. This measurement of the expression level of the biomarker (M1) of the reference substance (S) is recorded as N1 Ref.

[0077] In the context of the present invention, the reference expression level of a biomarker of aging (M2) can be obtained by measuring the expression level of the biomarker (M2):

[0078] - in a cell culture medium of "young" human dermal fibroblasts at passage R6 (the "young" cells at passage R6 were not brought into contact with senescent cells nor treated with the substance or chemical composition); this measurement of the expression level of the biomarker (M2) of the untreated and unstressed cells is recorded as N2, and / or

[0079] - in a cell culture medium obtained by contacting "young" human dermal fibroblasts obtained at passage R6 with aged human dermal fibroblasts obtained at passage R20; recording this measurement of the expression level of the marker (M2) of untreated but stressed cells as N20; and / or

[0080] - in the cell culture medium obtained at the end of the implementation of step b) of the method that is the subject of the invention, when the substance (S) used in step a) of the method that is the subject of the invention is a substance of an element chosen from the group consisting of vitamin E and vitamin C. This measurement of the expression level of the biomarker (M2) of the reference substance (S) is recorded as N2 Ref.

[0081] By means of the method as previously defined, said substance (S) or said composition (C) can be selected as an ingredient intended to prevent or slow down the appearance of or eliminate the signs of aging of human skin or lips and can be used as such or for the preparation of a composition for topical use in the cosmetic or dermo-cosmetic or pharmaceutical industry.

[0082] By means of the method as previously defined, the selected substance (S) or composition (C) is used as such or for the preparation of a composition for topical use comprising it, for preventing or slowing down the appearance of signs of aging on human skin or lips or for eliminating said signs, if:

[0083] - the expression level measured for said substance (S) or said composition (C), recorded as N1i, is lower than the reference expression level N10 of the biomarker (M1) of SASP / SMS, and if

[0084] - the expression level measured for said substance (S) or said composition (C), recorded as N2i, is lower than the reference expression level N20 of a biomarker of aging (M2).

[0085] More particularly the selected substance (S) or composition (C) is used as such or for the preparation of a cosmetic composition for topical use comprising it, for preventing or slowing down the appearance of signs of aging on human skin or lips or for eliminating said signs, if:

[0086] - the ratio R1 = [(N10-N1i) x 100] / [(N10-N1)] is greater than or equal to n1; and if

[0087] - the ratio R2 = [(N20-N2i) x 100] / [(N20-N2)] is greater than or equal to n2,

[0088] wherein n1 is greater than or equal to 15, and more particularly greater than or equal to 30, n2 is greater than or equal to 60, and more particularly greater than or equal to 70, and N1, N10, N2 and N20 are as previously defined, or

[0089] n1 as defined above is more particularly greater than or equal to 15, most particularly greater than or equal to 30 and even more particularly greater than or equal to 40; and n2 as defined above is more particularly greater than or equal to 60, most particularly greater than or equal to 70 and even more particularly greater than or equal to 80.

[0090] By means of the method as defined previously, the substance (S) or composition (C) will more particularly be selected if n1 is greater than or equal to 30 and if n2 is greater than or equal to 70.

[0091] A subject of the present invention is also a cosmetic composition (CA) comprising, per 100% of its mass, from 0.1% to 25% by mass of at least one compound of general formula (I):

[0092]

[0093] in:

[0094] -R1 represents a group having formula (Ia)

[0095] -CH2-CO2H(Ia),

[0096] and R2 represents a hydrogen atom, or

[0097] -R1 represents a group having formula (Ib)

[0098] -CH(CO2H)-CH2-CH2-OH (Ib),

[0099] and R2 represents a hydrogen atom, or

[0100] - R1 represents a group having formula (Ia), and R2 represents a hydroxyethyl group (Ic):

[0101] HO-CH2-CH2- (Ic), or

[0102] - R1 represents a group having formula (Ia), and R2 represents a hydroxymethylethyl group (Id):

[0103] HO-C(CH3)(CH2-CH3)(Id), or

[0104] - R1 represents a group having formula (Ia), and R2 represents a hydroxypropyl group (Ie):

[0105] HO-CH2-CH2-CH2-(Ie), or

[0106] - R1 represents a group having formula (Ia), and R2 represents an (E)-propenyl group (If), or

[0107] - R1 represents a group having formula (Ia), and R2 represents a (Z)-propenyl group (Ig), or

[0108] - R1 and R2 both represent a group having formula (Ia), or

[0109] -R1 represents a group having formula (Ii)

[0110] -CH(CO2H)-CH(OH)-CH3 (Ii)

[0111] and R2 represents a hydrogen atom, or

[0112] - R1 represents a group having formula (Ii), and R2 represents a methyl group, or

[0113] - R1 represents a group having formula (Ib), and R2 represents a methyl group, or

[0114] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Ij)

[0115] -CH(CO2H)-CH3(Ij), or

[0116] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Ib), or

[0117] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Ik)

[0118] -CH(CO2H)-CH(CH3)2 (Ik), or

[0119] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Ii), or

[0120] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Il)

[0121] -CH(CO2H)-CH2(CO2H) (Il), or

[0122] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Im)

[0123] -CH2-CH=CH-CO2H (Im).

[0124] According to a particular aspect, a subject of the invention is a cosmetic composition (CA) comprising, per 100% of its mass, from 0.1% to 25% by mass of at least one compound of general formula (I), in which:

[0125] - R1 and R2 both represent a group having formula (Ia), or

[0126] - R1 represents a group having formula (Ia), and R2 represents a hydrogen atom, or

[0127] - R1 represents a group having formula (Ia), and R2 represents a group (Ic), or

[0128] - R1 represents a group having formula (Ia), and R2 represents a group having formula (Ib), or

[0129] - R1 represents a group having formula (Ia), and R2 represents a group (If).

[0130] According to another particular aspect, a subject of the invention is a composition as defined previously, for preventing or slowing down the appearance of signs of aging or eliminating said signs on human skin or lips.

[0131] The cosmetic composition (CA) as defined above can be prepared by mixing the components in the desired mass ratios at room temperature or, if necessary, above the melting points of their ingredients and with appropriate mechanical stirring (50 to 150 rpm).

[0132] The cosmetic composition (CA) as defined above may also be prepared by any extraction technique known to a person skilled in the art and applied to red algae, more particularly to algae belonging to the phylum Rhodophyta, class Rhodophyceae, orders Acrochaetiales, Acrosymphytales, Ahnfeltiales, Balbianiales, Balliales, Batrachospermales, Bonnemaisoniales, Ceramiales, Colaconematales, Corallinales, Entwisleiales, Florideophyceae incertae sedis, Gelidiales, Gigartinales, Gracilariales, Halymeniales, Hildenbrandiales, Nemaliales, Nemastomatales, Palmariales, Peyssonneliales, Pihiellales, Plocamiales, Rhodachlyales, Rhodogorgonales, Rhodymeniales, Sebdeniales, Sporolithales, and Thoreals.

[0133] According to a more specific aspect, the cosmetic composition (CA) as defined above may also be prepared by any extraction technique known to a person skilled in the art and applied to red algae belonging to the order Bonnemaisoniales, the family Bonnemaisoniales, the genera / species Asparagopsis armata and Asparagopsis taxiformis.

[0134] A final subject of the present invention is a cosmetic formulation for topical use comprising at least one cosmetically acceptable excipient and an effective amount of a cosmetic composition (CA) as defined previously.

[0135] According to one particular aspect, the cosmetic formulation according to the invention will be a formulation for preventing or slowing down the appearance of or eliminating the signs of aging of the human skin or lips.

[0136] The cosmetic composition for topical use is generally spread on the surface of the skin to be treated, and then the skin is massaged for a while.

[0137] The expression "intended for topical use" as used in the definition of a cosmetic formulation means that the formulation is used by application to the skin, whether directly (in the case of a cosmetic formulation) or indirectly (when the cosmetic formulation according to the invention is impregnated on a support intended to come into contact with the skin (paper, wipes, textile, transdermal device, etc.)).

[0138] The term "cosmetically acceptable" as used in the definition of cosmetic formulations for topical use used in the method that is the subject of the present invention means any substance or formulation intended to come into contact with parts of the human body (epidermis, body hair and hair system, nails, lips and genitals) or with the teeth and oral mucosa for the exclusive and principal purpose of cleansing them, perfuming them, changing their appearance and / or correcting body odours and / or protecting them or keeping them in good condition, in accordance with EEC Council Directive 76 / 768 / EEC of 27 July 1976, as amended by Directive 93 / 35 / EEC of 14 June 1993.

[0139] The term "effective amount of composition (A) as defined previously and present in the cosmetic formulation for topical use that is the subject of the invention" means, per 100% by mass of said cosmetic formulation for topical use, a proportion of composition (A) of between 0.1% and 5% by mass, more particularly between 0.1% and 3% by mass, even more particularly between 0.5% and 2.5% by mass.

[0140] The cosmetic formulations for topical use according to the invention are generally in the form of aqueous or hydro-alcoholic or hydro-glycolic solutions, in the form of suspensions, emulsions, microemulsions or nanoemulsions, whether of the water-in-oil, oil-in-water, water-in-oil-in-water or oil-in-water-in-oil type, or in the form of powders.

[0141] The cosmetic preparations for topical use according to the invention can be packaged in bottles, in devices of the "pump bottle" type, in aerosol devices (in pressurized form), in devices equipped with perforated walls (such as grids) or in devices equipped with a ball applicator (known as a "roll-on").

[0142] Typically, the composition (CA) present in the cosmetic formulation for topical use according to the invention is combined with chemical additives usually used in the field of formulations for topical use, such as foaming and / or detergent surfactants, thickening and / or gelling surfactants, thickeners and / or gelling agents, stabilizers, film-forming compounds, solvents and co-solvents, hydrotropes, spring or mineral water, plasticizers, emulsifiers and co-emulsifiers, opacifiers, pearlescent agents, superfatting agents, chelating agents, complexing agents, oils, waxes, antioxidants, fragrances, essential oils, preservatives, conditioners, deodorants, whitening agents intended for bleaching body hair and skin, active ingredients intended for providing a treatment and / or protection effect on the skin or hair, sunscreens, mineral fillers or pigments, particles providing a visual effect or intended for encapsulating active agents, exfoliating particles, texturizing agents, optical whitening agents, or insect repellents.

[0143] As examples of foaming and / or detergent surfactants with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of anionic, cationic, amphoteric or nonionic foaming and / or detergent surfactants.

[0144] Among the foaming and / or detergent anionic surfactants, mention may be made of alkyl ether sulfates, alkyl sulfates, alkyl amido ether sulfates, alkyl aryl polyether sulfates, monoglyceride sulfates, α-olefin sulfonates, paraffin sulfonates, alkyl phosphates, alkyl ether phosphates, alkyl sulfonates, alkyl amide sulfonates, alkyl aryl sulfonates, alkyl carboxylates, alkyl sulfosuccinates, alkyl ether sulfosuccinates, alkylamide sulfosuccinates, alkyl sulfoacetates, alkyl sarcosinates, acyl isethionates, N-acyl taurates, acyl lactylates, N-acyl amino acid derivatives, N-acyl peptide derivatives, N-acyl protein derivatives, N-acyl derivatives, alkali metal salts, alkaline earth metal salts, ammonium salts, amine salts, or amino alcohol salts of fatty acids.

[0145] Among the foaming and / or detergent amphoteric surfactants, mention may be made of the alkyl betaines, alkylamidobetaines, sultaines, alkylamidoalkylsultaines, imidazoline derivatives, phosphobetaines, amphopolyacetates and amphopropionates.

[0146] Among the foaming and / or detergent cationic surfactants, mention may in particular be made of quaternary ammonium derivatives.

[0147] Among the foaming and / or detergent nonionic surfactants, mention may be made more particularly of alkyl polyglycosides comprising a linear or branched, saturated or unsaturated aliphatic group comprising from 8 to 16 carbon atoms, such as octyl polyglucoside, decyl polyglucoside, undecyl polyglucoside, dodecyl polyglucoside, tetradecyl polyglucoside, hexadecyl polyglucoside or 1,12-dodecanediyl polyglucoside; ethoxylated hydrogenated castor oil derivatives, such as the product sold under the INCI name PEG-40 Hydrogenated Castor Oil; polysorbates, such as polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 70, polysorbate 80 and polysorbate 85; cocoamide; N-alkylamines.

[0148] As examples of thickening and / or gelling surfactants that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may optionally be made of alkoxylated alkyl polyglycoside fatty esters, such as ethoxylated methyl polyglucoside esters, such as those known under the names Glutamate TM LT and Glutamate TM PEG 120 methyl glucose trioleate and PEG 120 methyl glucose dioleate sold as DOE 120; alkoxylated fatty esters such as those sold under the name Crothix TM DS53 is sold as PEG 150 Pentaerythritol Tetrastearate, also known as Antil TM PEG 55 propylene glycol oleate sold under the name Elfacos TM PPG-14 Laureth Isophoryl Diurethane sold by T211 under the name Elfacos TM PPG-14 Palmeth-60 Hexyl Dicarbamate sold by GT2125.

[0149] As examples of thickeners and / or gelling agents that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of polymers of the linear or branched or crosslinked polyelectrolyte type, such as partially or completely salified homopolymers of acrylic acid, partially or completely salified homopolymers of methacrylic acid, partially or completely salified homopolymers of 2-methyl[(1-oxo-2-propenyl)amino]-1-propanesulfonic acid (AMPS), copolymers of acrylic acid and AMPS, copolymers of acrylamide and AMPS, vinylpyrrolidone and AMPS. copolymers of AMPS and (2-hydroxyethyl) acrylate, copolymers of AMPS and (2-hydroxyethyl) methacrylate, copolymers of AMPS and hydroxyethyl acrylamide, copolymers of AMPS and N,N-dimethyl acrylamide, copolymers of AMPS and tris(hydroxymethyl)acrylamidomethane (THAM), copolymers of acrylic acid or methacrylic acid and (2-hydroxyethyl) acrylate, copolymers of acrylic acid or methacrylic acid and (2-hydroxyethyl) methacrylate, copolymers of acrylic acid or methacrylic acid and hydroxyethyl acrylamide , copolymers of acrylic acid or methacrylic acid and THAM, copolymers of acrylic acid or methacrylic acid and N,N-dimethylacrylamide, terpolymers of acrylic acid or methacrylic acid, AMPS and (2-hydroxyethyl)acrylate, terpolymers of acrylic acid or methacrylic acid, AMPS and (2-hydroxyethyl)methacrylate, terpolymers of acrylic acid or methacrylic acid, AMPS and THAM, terpolymers of acrylic acid or methacrylic acid, AMPS and N,N-dimethylacrylamide, terpolymers of acrylic acid or methacrylic acid, AMPS and acrylamide, copolymers of acrylic acid or methacrylic acid and alkyl acrylates whose carbon chain contains between 4 and 30 carbon atoms and more particularly between 10 and 30 carbon atoms, copolymers of AMPS and alkyl acrylates whose carbon chain contains between 4 and 30 carbon atoms and more particularly between 10 and 30 carbon atoms, linear, branched or crosslinked terpolymers of at least one monomer with free, partially salified or fully salified strong acid functional groups with at least one neutral monomer and at least one monomer having formula (VIII):

[0150] CH2=C(R'3)-C(=O)-[CH2-CH2-O] n -R'4(VIII)

[0151] wherein R′3 represents a hydrogen atom or a methyl group, R′4 represents a linear or branched alkyl group containing from 8 to 30 carbon atoms, and n represents a number greater than or equal to 1 and less than or equal to 50.

[0152] The polymers of the linear or branched or crosslinked polyelectrolyte type with which the composition (CA) according to the invention can be combined in the cosmetic formulation for topical use can be in the form of an aqueous solution, an aqueous suspension, a water-in-oil emulsion, an oil-in-water emulsion or a powder. The polymers of the linear or branched or crosslinked polyelectrolyte type with which the compound of formula (I) or the composition (C1) can be combined in the cosmetic formulation for topical use according to the invention can be chosen from the group consisting of the polymers of the type known under the name Simulgel TM EG, Simulgel TM EPG, Sepigel TM 305.Simulgel TM 600、Simulgel TM NS, Simulgel TM INS100、Simulgel TM FL, Simulgel TM A. Simulgel TM SMS 88, Sepinov TM EMT 10, Sepiplus TM 400、Sepiplus TM 265. Sepiplus TM S. Sepimax TM Zen, Aristoflex TM AVC、Aristoflex TM AVS, Novemer TM EC-1, Novemer TM EC 2, Aristoflex TM HMB, Cosmedia TM SP, Flocare TM ET 25, Flocare TM ET 75, Flocare TM ET 26, Flocare TM ET 30, Flocare TM ET 58, Flocare TM PSD 30, Viscolam TM AT 64 and Viscolam TM Products sold by AT 100.

[0153] As examples of thickeners and / or gelling agents that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of polysaccharides consisting only of monosaccharides, such as dextran or glucose homopolymers, glucomannan glucan, xyloglucan, galactomannans, wherein the degree of substitution (DS) of the D-galactose units on the main D-mannose chain is between 0 and 1, and more particularly between 1 and 0.25, such as galactomannans derived from cassia gum (DS=1 / 5), locust bean gum (DS=1 / 4), tara gum (DS=1 / 3), guar gum (DS=1 / 2) or fenugreek gum (DS=1).

[0154] As examples of thickeners and / or gelling agents with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of polysaccharides composed of monosaccharide derivatives, such as sulfated galactans and more particularly carrageenan and agar, uronans and more particularly algin, alginates and pectin, heteropolymers of monosaccharides and uronic acid, and more particularly xanthan gum, gellan gum, gum arabic exudate and gum karaya exudate, or glycosaminoglycans.

[0155] As examples of thickeners and / or gelling agents with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of cellulose, cellulose derivatives such as methylcellulose, ethylcellulose, hydroxypropylcellulose, silicates, starch, hydrophilic starch derivatives or polyurethanes.

[0156] As examples of stabilizers that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of microcrystalline waxes and more particularly ozokerite, mineral salts such as sodium chloride or magnesium chloride, silicone polymers such as polysiloxane polyalkyl polyether copolymers.

[0157] As examples of solvents that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of water, organic solvents such as glycerol, diglycerol, glycerol oligomers, ethylene glycol, propylene glycol, butylene glycol, 1,3-propanediol, 1,2-propanediol, hexylene glycol, diethylene glycol, xylitol, erythritol, sorbitol, water-soluble alcohols such as ethanol, isopropanol or butanol, or mixtures of water and said organic solvents.

[0158] As examples of thermal or mineral waters with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of thermal or mineral waters having a mineralization content of at least 300 mg / l, in particular Avene water, Vittel water, Vichy basin water, Uriage water, La Roche-Posay water, La Bourboule water, Enghien-les-Bains water, Saint-Gervais-les-Bains water, Néris-les-Bains water, Allevard-les-Bains water, Digne water, Maizières water, Neyrac-les-Bains water, Lons-le-Saunier water, Rochefort water, Saint-Gervais-les-Bains water, Néris-les-Bains water, Allevard-les-Bains water, Digne water, Maizières water, Neyrac-les-Bains water, Lons-le-Saunier water, Rochefort water, Saint-Gervais-les-Bains water, Christau water, Les Fumades water and Tercis-les-Bains water.

[0159] As examples of hydrotropes with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of xylene sulfonate, cumene sulfonate, hexyl polyglucoside, 2-ethylhexyl polyglucoside or n-heptyl polyglucoside.

[0160] As examples of emulsifying surfactants with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of nonionic surfactants, anionic surfactants or cationic surfactants.

[0161] As examples of emulsifying nonionic surfactants with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of fatty acid esters of sorbitol, for example those sold under the name Montane TM 40. Montane TM 60. Montane TM 70. Montane TM 80 and Montane TM 85; a composition comprising glyceryl stearate and stearic acid ethoxylated with between 5 mol and 150 mol of ethylene oxide, for example a composition comprising stearic acid ethoxylated with 135 mol of ethylene oxide and glyceryl stearate (marketed under the name Simulsol TM165); dehydrated mannitol esters, ethoxylated dehydrated mannitol esters; sucrose esters; methyl glucoside esters; alkyl polyglycosides containing linear or branched, saturated or unsaturated aliphatic groups and containing from 14 to 36 carbon atoms, such as tetradecyl polyglucoside, hexyldecyl polyglucoside, octadecyl polyglucoside, hexyldecyl polyxyloside, octadecyl polyxyloside, eicosyl polyglucoside, dodecosyl polyglucoside, 2-octyldodecyl polyxyloside, 12-hydroxystearyl polyglucoside; a combination containing from 14 to 36 carbon atoms, linear or branched, saturated or unsaturated aliphatic alcohols and an alkyl polyglycoside as described above, for example sold under the trade name Montanov TM 68. Montanov TM 14. Montanov TM 82. Montanov TM 202. Montanov TM S. Montanov TM WO18, Montanov TM L. Fluidanov TM 20X and Easynov TM Composition for sale.

[0162] As examples of anionic surfactants with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of glyceryl stearate citrate, cetearyl sulfate, soaps such as sodium stearate or triethanolammonium stearate, or salified N-acyl derivatives of amino acids (for example stearoyl glutamate).

[0163] As examples of emulsifying cationic surfactants that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of ammonium oxides, quaternium-82 and the surfactants described in patent application WO 96 / 00719, and mainly those whose fatty chain contains at least 16 carbon atoms.

[0164] As examples of sunscreens and / or pearlescent agents with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of sodium palmitate, sodium stearate, sodium hydroxystearate, magnesium palmitate, magnesium stearate, magnesium hydroxystearate, ethylene glycol monostearate, ethylene glycol distearate, polyethylene glycol monostearate, polyethylene glycol distearate, and fatty alcohols comprising from 12 to 22 carbon atoms.

[0165] As examples of texturizing agents with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of N-acylamino acid derivatives, for example those sold under the name Aminohope TM Lauroyl lysine sold by LL under the name Dryflo TM Octenyl starch succinate is sold under the name Montanov TM 14 Myristyl polyglucoside, cellulose fiber, cotton fiber, chitosan fiber, talc, sericite and mica for sale.

[0166] As examples of deodorants that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of alkaline silicates, zinc salts such as zinc sulfate, zinc gluconate, zinc chloride or zinc lactate; quaternary ammonium salts, for example cetyltrimethylammonium salts, or cetylpyridinium salts; glycerol derivatives, for example glyceryl caprate, glyceryl caprylate and polyglyceryl caprate; 1,2-decanediol, 1,3-propylene glycol; salicylic acid; sodium bicarbonate; cyclodextrins; metal zeolites; Triclosan TM ; hydrated aluminum bromide, hydrated aluminum chlorohydrate, aluminum chloride, aluminum sulfate, hydrated aluminum zirconium chlorohydrate, hydrated aluminum zirconium trichloride, hydrated aluminum zirconium tetrachloride, hydrated aluminum zirconium pentachloride, hydrated aluminum zirconium octachloride, aluminum sulfate, sodium aluminum lactate, a complex of hydrated aluminum chlorohydrate and a glycol, such as a complex of hydrated aluminum chlorohydrate and propylene glycol, a complex of hydrated aluminum dichloride and propylene glycol, a complex of sesquichlorohydroxyaluminum and propylene glycol, a complex of hydrated aluminum chlorohydrate and polyethylene glycol, a complex of hydrated aluminum dichloride and polyethylene glycol, or a complex of sesquichlorohydroxyaluminum and polyethylene glycol.

[0167] As examples of oils that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of mineral oils, such as liquid paraffin, liquid vaseline, isoparaffin or white mineral oil; oils of animal origin, such as squalene or squalane; vegetable oils, such as squalane, sweet almond oil, coconut oil, castor oil, jojoba oil, olive oil, rapeseed oil, peanut oil, sunflower oil, wheat germ oil, corn germ oil, soybean oil, cottonseed oil, alfalfa oil, poppy oil, pumpkin oil, evening primrose oil, millet oil, barley oil, rye oil, safflower oil, tung oil, passionflower oil, hazelnut oil, palm oil, shea butter, almond oil, sedge leaf oil, garlic mustard oil, avocado oil, calendula oil, oils derived from flowers or vegetables, ethoxylated vegetable oils; synthetic oils, for example fatty acid esters, such as butyl myristate, propyl myristate, isopropyl myristate, cetyl myristate, palm oil, Isopropyl palmitate, octyl palmitate, butyl stearate, cetyl stearate, isopropyl stearate, octyl stearate, isocetyl stearate, dodecyl oleate, hexyl laurate, propylene glycol dicaprylate, esters derived from lanolin acid (such as isopropyl lanolinate, isocetyl lanolinate), fatty acid monoglycerides, diglycerides and triglycerides (such as triheptanoin), alkyl benzoates, hydrogenated oils, poly(α-olefins ), polyolefins (such as poly(isobutane)), synthetic isoalkanes (such as isohexadecane, isododecane), perfluorinated oils; silicone oils, such as dimethylpolysiloxane, methylphenylpolysiloxane, silicones modified with amines, silicones modified with fatty acids, silicones modified with alcohols, silicones modified with alcohols and fatty acids, silicones modified with polyether groups, silicones modified with epoxy groups, silicones modified with fluorine groups, cyclic silicones, and silicones modified with alkyl groups. In the present patent application, the term "oil" refers to a compound and / or a mixture of compounds that is water-insoluble and has a liquid appearance at a temperature of 25°C.

[0168] As examples of waxes with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of beeswax, carnauba wax, candelilla wax, carnauba wax, Japan wax, cork fiber wax, sugar cane wax, paraffin wax, montan wax, microcrystalline wax, lanolin wax; ozokerite; polyethylene wax; silicone wax; vegetable wax; fatty alcohols and fatty acids that are solid at room temperature; glycerides that are solid at room temperature. In the present patent application, the term "wax" refers to a compound and / or a mixture of compounds that is water-insoluble and has a solid appearance at a temperature greater than or equal to 45°C.

[0169] As examples of active ingredients with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of vitamins and their derivatives, in particular their esters such as retinol (vitamin A) and its esters such as retinyl palmitate, ascorbic acid (vitamin C) and its esters, sugar derivatives of ascorbic acid such as ascorbyl glucoside, tocopherol (vitamin E) and its esters such as tocopheryl acetate, vitamin B3 or B10 (niacinamide and its derivatives); compounds which exhibit a whitening or depigmenting effect on the skin, such as those sold under the name Sepiwhite TM MSH, Sepicalm TM Omega-undecenoylphenylalanine, mono- and / or diglycerides of omega-undecenoylphenylalanine, omega-undecenoyl dipeptide, arbutin, kojic acid, hydroquinone; compounds showing sedative effects, in particular Sepicalm TM S, allantoin and bisabolol; anti-inflammatory agents; compounds showing moisturizing effects, such as urea, hydroxyurea, glycerol, polyglycerol, glycerol glucoside, diglycerol glucoside, polyglycerol glucoside, xylitol glucoside; plant extracts rich in polyphenols, such as grape extract, pine extract, wine extract and olive extract; compounds showing anti-obesity or lipolysis effects, such as caffeine or its derivatives, Adiposlim TM 、Adipoless TM , fucoxanthin; N-acyl proteins; N-acyl peptides such as Matrixyl TM ; N-acyl amino acids; N-acyl partial protein hydrolysates; amino acids; peptides; total protein hydrolysates; soy extracts, such as Raffermine TM ; Wheat extracts, such as Tensine TM or Gliadine TM ; plant extracts, such as tannin-rich plant extracts, isoflavone-rich plant extracts or terpene-rich plant extracts; freshwater or marine algae extracts; marine plant extracts; marine extracts, typically coral; essential wax; bacterial extracts; ceramides; phospholipids; compounds showing antimicrobial or purifying effects, such as Lipacide TM C8G、Lipacide TM UG, Sepicontrol TM A5; Octopirox TM or Sensiva TM SC50; Compounds that display stimulatory or stimulating properties, such as Physiogenyl TM , panthenol and its derivatives such as Sepicap TMMP; Anti-aging actives, such as Sepilift TM DPHP、Lipacide TM PVB, Sepivinol TM , Sepivita TM , Manoliva TM 、Phyto-Age TM 、Timecode TM ;Survicode TM ; anti-photoaging actives; agents for protecting the integrity of the dermal-epidermal junction; agents for increasing the synthesis of components of the extracellular matrix, such as collagen, elastin and glycosaminoglycans; actives that act favorably on chemical cell communication, such as cytokines, or on physical cell communication, such as integrins; actives that produce a "warming" sensation on the skin, such as activators of microcirculation (such as nicotinic acid derivatives) or products that produce a "cooling" sensation on the skin (such as menthol and its derivatives); actives that improve skin microcirculation, such as venous tension agents; drainage actives; decongestant actives, such as Ginkgo, Ivy, Horse Chestnut, Bamboo, Ruscus, Butcher's extracts of safflower, centella, asiatica, rosemary or willow; tanning or darkening agents, for example dihydroxyacetone (DHA), erythrulose, mesotartaric aldehyde, glutaraldehyde, glyceraldehyde, alloxan or ninhydrin, plant extracts, for example extracts of rosewoods of the genera Pterocarpus and Baphia, for example Pteropcarpus santalinus, Pterocarpus osun, Pterocarpus soyauxii, Pterocarpus erinaceus, Pterocarpus indicus or Baphia nitida, as disclosed in European Patent Application EP 0 971 683; agents known for their effect of promoting and / or accelerating the tanning or darkening of human skin and / or for their effect of pigmenting human skin, such as carotenoids (and more particularly beta-carotene and gamma-carotene), the product sold under the trade name "Carros Oil" (INCI name: Daucus carrota, Helianthus annuus oil) by the company Provital, which contains carotenoids, vitamin E and vitamin K; tyrosine and / or its derivatives, known for their effect of accelerating the tanning of human skin in combination with exposure to ultraviolet radiation, for example sold under the trade name SunTanAccelerator by the company Provital TMA product sold under the trade name MelanoBronze by Mibelle that contains tyrosine and riboflavin (vitamin B), a tyrosine and tyrosinase complex sold under the trade name Zymo Tan Complex by Zymo Line, and a tyrosinase complex sold under the trade name MelanoBronze by Mibelle. TM (INCI name: Acetyl Tyrosine, Monk's Pepper Extract (Vitex agnus-castus)), which contains acetyl tyrosine, a product sold by the company Unipex under the trade name Unipertan VEG-24 / 242 / 2002 (INCI name: Butylene Glycol and Acetyl Tyrosine with Hydrolyzed Vegetable Protein and Adenosine Triphosphate), a product sold by the company Sederma under the trade name Try-Excell TM A product sold under the trade name Actibronze TM (INCI name: Hydrolyzed wheat protein with acetyl tyrosine and copper gluconate), marketed by Synerga under the brand name Tyrostan TM (INCI name: Potassium Caproyl Tyrosinate), the product sold by Synerga under the brand name Tyrosinol (INCI name: Sorbitan Isostearate, Glyceryl Oleate, Caproyl Tyrosine), the product sold by Alban Muller under the brand name InstaBronze TM products sold under the brand name Tyrosilane (INCI name: Methylsilanol and Acetyl Tyrosine) by the company Exymol; peptides known for their melanogenesis-activating effect, such as the product sold under the brand name Bronzing SF Peptide Powder (INCI name: Glucan and Octapeptide-5) by the company Infinitec Activos, the product sold under the brand name Melitane (INCI name: Glycerin and Water and Glucan and Acetyl Hexapeptide-1), which contains Acetyl Hexapeptide-1 known for its α-MSH agonist effect, and the product sold under the brand name Melatimes Solutions by the company Lipotec TM Products sold under the trade name Tanositol TM(INCI name: Inositol), sold by CODIF International under the trade name Thalitan TM (or Phycosaccharide TM A product sold by Alban Muller AG (INCI name: Water and hydrolyzed alginate (Laminaria digitata) and magnesium and manganese sulfates) containing oligosaccharides of marine origin (guluronic acid and mannuronic acid chelated with magnesium and manganese ions) under the trade name Melactiva TM products sold under the trade name “Biotanning” (INCI name: hydrolyzed sweet orange (citrus Aurantium dulcis) fruit extract), compounds rich in flavonoids, such as the product sold under the trade name “Biotanning” (INCI name: hydrolyzed sweet orange (citrus Aurantium dulcis) fruit extract) by the company Silab, which is known to be rich in citric flavonoids (hesperidins); agents intended for treating hair and / or body hair, such as agents for protecting hair follicle melanocytes, intended to protect said melanocytes from cytotoxic agents that are the cause of senescence and / or apoptosis of said melanocytes, such as those selected from those described in the European patent application published under EP 1 515 688 A2, mimetics of dopachrome tautomerase activity, synthetic SOD mimetic molecules, such as manganese complexes, antioxidant compounds, such as cyclodextrin derivatives, silicon-containing compounds derived from ascorbic acid, lysine or arginine pyrrolidonecarboxylates, combinations of mono- and diesters of cinnamic acid and vitamin C, and more generally the compositions described in EP 1 515 688 A2. Those mentioned in European patent application published under No. 1 515 688 A2.

[0170] As examples of antioxidants with which the composition (CA) in the cosmetic formulation for topical use according to the invention can be combined, mention may be made of EDTA and its salts, citric acid, tartaric acid, oxalic acid, BHA (butylated hydroxyanisole), BHT (butylated hydroxytoluene), tocopherol derivatives such as tocopherol acetate, mixtures of antioxidant compounds such as Dissolvine sold under the INCI name by the company AkzoNobel GL 47S: Tetrasodium Glutamate Diacetate).

[0171] As examples of sunscreens that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of all those that appear in the amended Cosmetics Directive 76 / 768 / EEC, Annex VII. Among the organic sunscreens that can be combined with the compound of formula (I) or the composition (C1) in the cosmetic formulation for topical use according to the invention, mention may be made of the family of benzoic acid derivatives, such as para-aminobenzoic acid (PABA), in particular the monoglyceride of PABA, the ethyl ester of N,N-propoxy PABA, the ethyl ester of N,N-diethoxy PABA, the ethyl ester of N,N-dimethyl PABA, the methyl ester of N,N-dimethyl PABA and the butyl ester of N,N-dimethyl PABA; the family of anthranilic acid derivatives, such as N-acetylanthranilate; the family of salicylic acid derivatives, such as amyl salicylate, homomenthyl salicylate, salicylate), ethylhexyl salicylate, phenyl salicylate, benzyl salicylate and p-isopropylphenyl salicylate; the family of cinnamic acid derivatives, such as ethylhexyl cinnamate, ethyl-4-isopropyl cinnamate, methyl 2,5-diisopropyl cinnamate, p-methoxypropyl cinnamate, p-methoxyisopropyl cinnamate, p-methoxyisoamyl cinnamate, p-methoxyoctyl cinnamate (p-methoxy-2-ethylhexyl cinnamate), p-methoxy-2-ethoxyethyl cinnamate, p-methoxycyclohexyl cinnamate, ethyl-α-cyano-β-phenyl cinnamate, 2-ethylhexyl- α-Cyano-β-phenyl ester or cinnamic acid glyceryl di-p-methoxymono-2-ethylhexanoyl ester; the family of benzophenone derivatives, such as 2,4-dihydroxybenzophenone, 2,2'-dihydroxy-4-methoxybenzophenone, 2,2',4,4'-tetrahydroxybenzophenone, 2-hydroxy-4-methoxybenzophenone, 2-hydroxy-4-methoxy-4'-methylbenzophenone, 2-hydroxy-4-methoxybenzophenone-5-sulfonate, 4-phenylbenzophenone, 2-ethylhexyl 4'-phenyl Benzophenone-2-carboxylate, 2-hydroxy-4-n-octyloxybenzophenone, 4-hydroxy-3-carboxybenzophenone; 3-(4'-methylbenzylidene)-d,l-camphor, 3-(benzylidene)-d,l-camphor, camphor benzalkonium methylsulfate; urocanic acid, ethyl urocanate; the family of sulfonic acid derivatives, such as 2-phenylbenzimidazole-5-sulfonic acid and its salts; the family of triazine derivatives, such as hydroxyphenyl triazine, ethylhexyloxyhydroxyphenyl-4-methoxyphenyl triazine, 2,4,6-triphenylamine 4,4-((6-(((1,1-dimethylethyl)amino)carbonyl)phenyl)amino)-1,3,5-triazine-2,4-diyldiimino)bis(2-ethylhexyl)benzoate, 2-phenyl-5-methylbenzoxazole, 2,2'-hydroxy-5-methylphenylbenzotriazole, 2-(2'-hydroxy-5'-tert-octylphenyl)benzotriazole, 2-(2'-hydroxy-5'-tert-octylphenyl)benzotriazole, -methylphenyl) benzotriazole; dibenzazine; di-p-methoxybenzoylmethane, 4-methoxy-4"-tert-butylbenzoylmethane; 5-(3,3-dimethyl-2-norbornene)-3-pentan-2-one; the family of diphenylacrylate derivatives, such as 2-ethylhexyl-2-cyano-3,3-diphenyl-2-acrylate, 2-cyano-3,3-diphenyl-2-acrylate ethyl ester; the family of polysiloxanes, such as benzylidenesiloxane malonate.

[0172] Among the mineral sunscreens (also called "mineral sunscreens") that can be combined with the composition (CA) in the cosmetic formulation for topical use according to the invention, mention may be made of titanium oxide, zinc oxide, cerium oxide, zirconium oxide, yellow, red or black iron oxides, and chromium oxide. These mineral sunscreens may or may not be micronized, may or may not have undergone a surface treatment and may optionally be in the form of an aqueous or oily predispersion.

[0173] The following experimental part illustrates the invention without however limiting it.

[0174] A-1)-Choice of selection method

[0175] A-1-1) Selection of senescence induction type

[0176] Four different types of senescence induction were studied:

[0177] - Acute exposure of "young" fibroblasts obtained at passage R6 to UV-A radiation; several radiation doses were tested: 5, 10, 15 or 20 J / cm2, with one exposure per day;

[0178] - Chronic exposure of "young" fibroblasts obtained at passage R6 to UV-A radiation; several radiation doses were tested: 3x 5J / cm2 or 6x 2.5J / cm2, twice daily exposures;

[0179] - Use of "artificially aged" fibroblasts that have been passaged by serial replication up to R18;

[0180] - Use of fibroblast banks obtained from elderly donors (average age 58 years)

[0181] It should be noted that both young fibroblasts (R6 passage) and aged fibroblasts (R20 passage) were obtained from the same donor and the same batch of cells. Fibroblasts obtained at R20 passage were obtained after serial replication of R6 cells.

[0182] To evaluate the most appropriate type of induction, fibroblasts (whether obtained at passage R6 or passage R20) were seeded at 15,000 or 10,000 cells / well in a 48-well plate in standard culture medium for fibroblasts at 37°C in a humidified atmosphere containing 5% CO2.

[0183] Each condition was performed in quadruplicate.

[0184] Infectious aging or bystander effect was then assessed by measuring the amount of extracellular IL-6 and the activity of β-galactosidase:

[0185] - For chronic and acute UVA exposure

[0186] - For fibroblasts obtained from elderly donors, the cells were collected at 96h and 144h.

[0187] - For replication-induced senescence, at 24h and 48h.

[0188] The results obtained were then allowed to calculate the amount of extracellular IL-6 and the degree of change in β-galactosidase activity, and to collect qualitative observations; all of these results are collated in Table 1 below.

[0189]

[0190] Table 1

[0191] Only when fibroblasts replicated up to passage R20, it was possible to observe increased IL-6 secretion and an increase in β-galactosidase, characteristic of a phenotype marking infectious aging or a bystander effect.

[0192] Therefore, the above tests made it possible to select continuous replication as the type of senescence induction used to mimic the model of infectious aging.

[0193] A-1-2) Selection of the most suitable model for simulating infectious aging

[0194] Use multiple methods to choose which one is best suited to observe the "bystander effect" on fibroblasts harvested at passages R6 and R20:

[0195] - Indirect co-culture (ready-to-use inserts) of R6 young fibroblasts and R20 aged fibroblasts in standard culture medium for fibroblasts

[0196] - These cells were co-cultured directly in the same well, where senescent cells obtained at passage R20 were labeled to distinguish them

[0197] - MCR20 medium conditioned with R20 senescent fibroblasts was used in contact with "young" fibroblasts obtained at passage R6.

[0198] Description of the protocol used to determine the most appropriate model for assessing infectious aging:

[0199] Indirect co-culture model using inserts:

[0200] Normal human fibroblasts obtained at passages R5 and R19 were expanded until reaching passages R6 and R20, respectively, and then R6 cells were seeded at 30 000 cells / well in 24-well plates, and R20 cells were seeded at 15 000 cells / well in inserts.

[0201] After 72 h of expansion in inserts and 24-well plates, inserts containing R20 cells were deposited in 24-well plates containing R6 cells and allowed to come into contact for 24 h and 4 days.

[0202] After 24 h and 4 days of incubation, the following senescence markers were assessed:

[0203] -Assessment of β-galactosidase activity by staining

[0204] - Evaluation of interleukin-6 by ELISA

[0205] Direct co-culture model:

[0206] Normal human fibroblasts obtained at passages R5 and R19 were expanded until reaching passages R6 and R20, respectively, and then seeded into culture dishes at 0.3 M / dish.

[0207] After 5 days of expansion in culture dishes, the cell lawn of cells at passage R20 was fluorescently labeled (Alexa Fluor at 5 mM). 488 NHS Ester marker). They were then seeded into 48-well plates at a 1 / 1 ratio with non-senescent and unlabeled R6 cells (i.e., two passages, 7500 cells / well). The cells thus seeded were then incubated for 48 hours.

[0208] After 48 h of incubation, the following senescence markers were assessed:

[0209] -Assessment of β-galactosidase activity by staining

[0210] - Evaluation of interleukin-6 by ELISA

[0211] Models using conditioned media:

[0212] Normal human fibroblasts were obtained at passage R19 and expanded until passage R20, and then seeded into culture dishes at 0.3 M / dish.

[0213] After 4 days of expansion in the culture dishes, the medium of all culture dishes was changed and then recovered after 48 hours of incubation. This recovered medium corresponds to the medium conditioned by senescent cells containing SMS medium (MCR20).

[0214] In parallel, R6 "young" fibroblasts were seeded at 10,000 and 15,000 cells / well in 48-well plates. After 72 hours of incubation, MCR20 conditioned medium was deposited on these cells. The cells thus treated with the medium conditioned by senescent cells were incubated for 24 and 48 hours.

[0215] The following aging markers were then assessed:

[0216] -Assess β-galactosidase activity by staining at 48 hours

[0217] - Assessment of interleukin-6 levels by ELISA at 24 hours

[0218] in conclusion

[0219] The use of conditioned media revealed conclusive and reliable results for the only model of infectious aging: an increase in interleukin-6 and a change in its metabolic activity (increase in β-galactosidase activity). These changes are characteristic of aging in "young" cells treated with media conditioned from aged cells.

[0220] A-2)-Definition of best practices

[0221] Therefore, the specific method includes the following steps:

[0222] - Step i ) Human dermal fibroblasts obtained at passage R19 were cultured in T25 flasks,

[0223] - Step ii ) amplified in Step i) The cells obtained at the end of the experiment were used to obtain medium conditioned by senescent cells or "MC R20".

[0224] The cells obtained at the end of step i) are expanded by seeding the cells in culture dishes at 0.3 M / dish. After 4 days of expansion in the dishes, the medium of all the dishes is replaced and then recovered after 48 hours of incubation. This recovered medium corresponds to the medium conditioned by senescent cells containing SMS medium (MCR20).

[0225] - Step iii ) Human dermal fibroblasts obtained at passage R5 were cultured in T25 flasks,

[0226] - Step iv) Amplification Step iii) cells obtained at the end of the expansion phase to obtain "young" or "R6" cells,

[0227] Steps iii) and iv) are carried out in parallel with steps i) and ii).

[0228] The cells obtained at the end of step iii) were expanded by seeding said cells at 10 000 and 15 000 cells / well in 48-well plates.

[0229] - Step v) The chemical substance (S) or chemical composition (C) is combined with Step iv) Contact with cell culture medium obtained in;

[0230] - Step vi) Make Step ii) The MCR20e conditioned medium obtained in Step iii) The "young" or "R6" cells in the expansion phase obtained in

[0231] In this step d), the MCR20 conditioned medium obtained in step ii) is brought into contact with the "young" or "R6" cells obtained in step v) for 48 hours.

[0232] - Step vii) The amount of extracellular interleukin IL-6 present in the culture supernatant is measured, and the β-galactosidase activity present in the cells obtained from step vi) is measured.

[0233] - Step viii)Comparison of extracellular IL-6 and β-galactosidase expression levels

[0234] More precisely, the following is calculated in the context of this step viii):

[0235] The ratio of extracellular interleukin IL-6 is R1=[(N10-N1i)x 100] / [(N10-N1)], where:

[0236] - step v) or step vi) of the method was not carried out (cells at passage R6, not treated with and not combined with conditioned cells of passage R20), N1 corresponds to the amount of IL-6 measured in the culture medium obtained at the end of the method as described above,

[0237] - step v of the method was not carried out) (cells at passage R6, not treated with conditioned cells of passage R20 but combined therewith), N10 corresponds to the amount of IL-6 measured in the culture medium obtained at the end of the method as described above,

[0238] - When substance or composition (i) is used in step v) of the method (cells at passage R6, not treated with and not combined with conditioned cells at passage R20), N1i corresponds to the amount of IL-6 measured in the culture medium obtained at the end of the method as described above.

[0239] The ratio of extracellular interleukin IL-6 is R2 = [(N20-N2i) x 100] / [(N20-N2)], where:

[0240] - not subjected to step v) or step vi) of the method) (cells at passage R6, not treated with and not combined with conditioned cells of passage R20), N2 corresponds to the amount of β-galactosidase measured in the culture medium obtained at the end of the method as described above,

[0241] - step v of the method was not carried out) (cells at passage R6, not treated with conditioned cells of passage R20 but combined therewith), N20 corresponding to the amount of β-galactosidase measured in the culture medium obtained at the end of the method as described above,

[0242] - When substance or composition (i) is used in step v) of the method (cells at passage R6, not treated with and not combined with conditioned cells of passage R20), N2i corresponds to the amount of β-galactosidase measured in the culture medium obtained at the end of the method as described above.

[0243] B]- Evaluation of substances and compositions according to the method which is the subject of the invention

[0244] The composition (CA1) was prepared by extracting the pre-dried and screened red alga Asparagaceae spinosa powder with a 1,3-propylene glycol / water mixture (40 / 60 v / v) to obtain an Asparagaceae spinosa extract at 1.2% of the solvent.

[0245] The composition (CA2) was prepared by maceration in ethanol at a temperature of 20° C. for a period of 2 hours. The extract thus obtained was evaporated and taken up in methanol, which precipitated the minerals present, which were removed by filtration. The filtrate was then evaporated and then analyzed by semipreparative HPLC (Phenomenex TM C18 column) and the collected fractions were saved and then analyzed to determine the amount of mycosporin-like amino acids at a palythine equivalent ratio of 50 mg / L.

[0246] After implementing the previously described method, the results obtained for each composition are summarized in the following Table 2:

[0247] Compositions (CA1) and (CA2) were then diluted in 1,3-propylene glycol to a ratio of 0.1% by mass.

[0248] Table 2

[0249]

[0250] (*): Vitamin E is D-α-tocopheryl acetate

[0251] Analysis of results

[0252] The ratio R1 is greater than 30 both for vitamin C used as reference compound and for compositions (CA1) and (CA2) according to the invention.

[0253] The ratio R1 is greater than 70 both for vitamin E used as reference compound and for compositions (CA1) and (CA2) according to the invention.

[0254] These compositions (CA1) and (CA2) may be selected to prevent infectious aging or bystander effects.

[0255] Bibliography

[0256] [1]: Campisi, J. and d'Adda di Fagagna F., "Cellular senescence: when bad things happen to good cells." Nat. Rev., Mol. Cell Biol., 2007, 8, 729-740

[0257] [2]:Coppe J.P.et al.,“Senescence-associated secretory phenotypesreveal cell-nonautonomous functions of oncogenic RAS and the p53 tumorsuppressor”,PloS Biol.,2008,6,2853-2868

[0258] [3]:Passos J.F.et al.,“A feedback between p21 and reactive oxygenproduction is necessary for cell senescence”,Mol.Syst.Biol,2010,6,347

[0259] [4]:Nelson G.,Wordsworth J.,Wang C.,Jurk D.,Lawless C.,Martin-RuizC.and Von Zglinicki T.:“A senescent cell bystander effect:senescence-inducedSenescence”.Aging Cell,2012,11,345-349.

[0260] [5]:Redmond R.W.,Rajadurai A.,Udayakumar D.,Sviderskaya E.V.,Tsao H.“Melanocytes are selectively vulnerable to UVA-mediated Bystander OxidativeSignaling.”J.Invest.Dermatol.2014,134,pp.1083-1090.

Claims

1. A method for evaluating the ability of a chemical substance or chemical composition to prevent or slow the appearance of or eliminate signs of aging on human skin or lips; the method comprising: - step a) contacting a chemical substance or a chemical composition with "young" human dermal fibroblasts at passage R6 obtained from a culture medium, wherein these "young" human dermal fibroblasts at passage R6 obtained from a culture medium are obtained by expansion of "young" human dermal fibroblasts at passage R5 obtained from a culture medium, - step b) contacting human senescent dermal fibroblasts with "young" fibroblasts obtained from step a), wherein the human senescent dermal fibroblasts are human dermal fibroblasts at passage R20 obtained from the culture medium or human dermal fibroblasts at passage R19 obtained from the culture medium before undergoing expansion, and - Step c) measuring the entry of "young" fibroblasts into senescence and comparing with a reference value, comprising using at least two biomarkers, wherein the biomarkers comprise: - a biomarker (M1) of the senescence-associated secretory phenotype (SASP) / senescence-message secretome (SMS), which is interleukin-6, and - a biomarker of aging (M2), which is β-galactosidase, Wherein, in step c), the expression levels of the two biomarkers (M1) and (M2) are compared with at least one reference expression level of each of the two biomarkers.

2. The method according to claim 1, characterized in that These people have signs of aging on their skin or lips selected from - the appearance of wrinkles, fine lines or microtopographical damage; and / or - the appearance of inflexibility and / or nervousness; and / or - Lack of density and / or appearance of firmness.

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