ECO-BIOLOGICAL COSMETIC COMPOSITION CAPABLE OF LONG-LASTINGLY REDUCING THE SIGNS OF SKIN AGING

Abstract

The present invention relates to a cosmetic composition, advantageously ecobiological, comprising: - at least one bacterioruberin; - at least one active form of proline selected from the group consisting of dipalmitoyl hydroxyproline, hydroxyproline, acetyl proline, amidinoproline, cocoyl proline, dihydroxypropyl proline, proline, N-acetylhydroxyproline, and their salts. [Fig. 1]

Description

Title of the invention: ECO-BIOLOGICAL COSMETIC COMPOSITION CAPABLE OF LONG-LASTINGLY REDUCING THE SIGNS OF SKIN AGING FIELD OF INVENTION

[0001] The present invention relates to the field of cosmetics, in particular to cosmetic compositions, advantageously ecobiological, and their uses to prevent and / or combat the appearance of signs of skin aging or to ensure or promote skin regeneration, in particular skin healing. PRIOR TECHNOLOGY

[0002] The skin is the heaviest and largest organ of the human body, composed of three main layers, from deepest to most superficial: the hypodermis, the dermis, and the epidermis. This organ constitutes the main barrier that prevents water loss from the body and separates the body from the external environment to protect against various physical, chemical, and microbial aggressions from the surrounding environment.

[0003] The epidermis, in contact with the external environment, is a multi-layered, keratinized, squamous, avascular epithelium; it is mainly composed of layers of keratinocytes, which represent 90 to 95% of the epidermal cells; it also contains melanocytes, Langerhans cells, and Merkel cells. In this tissue, the extracellular compartment is reduced to the intercellular cement.

[0004] Conversely, the dermis consists of a small number of cells and a well-developed matrix. Fibroblasts constitute the majority of resident cells in the dermis. They are dispersed throughout the extracellular matrix, a polymer network composed primarily of proteins, notably collagen and elastin, as well as glycoproteins, proteoglycans (PGs), and glycosaminoglycans (GAGs). The dermis is traversed by a branching network of blood vessels, lymphatic vessels, and nerves that connect the skin to the rest of the body.

[0005] The hypodermis is the deepest and thickest part of the skin (up to 3 cm thick on the abdomen); it is a major area of ​​fatty deposits. The hypodermis separates the skin from the underlying tissues.

[0006] It is known that the signs of skin aging result in changes in the skin's condition that are essentially due to a slowing down and imbalance of skin function. This is accompanied by a decrease in the quality of the dermis with a drop in the levels of structural matrix proteins, e.g., PG and GAG, accompanied by a loss of consistency of the extracellular matrix; a decrease in the thickness of the epidermis (slowing of cell renewal or hypoplasia), a decrease in the production of lipids and epidermal structural proteins, an imbalance of desquamation as well as a reduction in the water content of the skin.

[0007] The most obvious manifestations of skin aging, particularly of the face, are the formation of wrinkles and fine lines, the appearance of pigment spots, and the loss of radiance and density of the skin.

[0008] A currently proposed solution for masking superficial skin imperfections is the use of fillers or so-called "blurring" agents such as talc, starch, mica, silica, poly-beta-alanine, sericite, brown iron oxide, or titanium dioxide. However, these compounds, which are generally in powder form, only provide a visual correction of surface imperfections (e.g., wrinkles, fine lines, or enlarged pores) through an optical / visual smoothing effect on the skin's surface with light-diffusing agents whose sole purpose is to fill in the imperfections. Thus, the skin is visually unified and mattified, and the appearance of wrinkles / fine lines and enlarged pores is reduced by a process akin to "camouflage." This solution is not long-lasting, particularly because these compounds tend to migrate into wrinkles during the day, but also because of makeup removal at the end of the day.

[0009] Another approach involves applying cosmetic compositions capable of biologically combating skin aging, promoting skin cell regeneration, and thus potentially aiding the healing of damaged skin, particularly in the elderly. These various applications are generally grouped under the heading "anti-aging." These products incorporate active ingredients to directly or indirectly stimulate fibroblast growth, strengthen the dermo-epidermal junction, combat oxidative stress, and promote the production of structural proteins and glycans, thereby promoting skin regeneration and repair.

[0010] Among the prior art solutions, many compounds are not free from deleterious side effects. For example, retinoids are described for their ability to stimulate fibroblast growth. However, their use is not recommended in cosmetics due to their phototoxic and potentially mutagenic action (Mei et al., 2005).

[0011] Furthermore, in most cases, so-called anti-aging cosmetic compositions have a rapid positive effect on the signs of skin aging, often linked to improved skin hydration. However, the long-term benefits of applying these anti-aging compositions are negligible. It is clear from what precedes that there remains a need to offer an effective anti-aging solution that provides long-term beneficial effects.

[0012] Prior art solutions that offer long-term effects include invasive procedures such as dermabrasion, injections of fillers or botulinum toxin, laser treatment, and other forms of cosmetic surgery. While these solutions reduce the signs of skin aging more permanently, they also present significant risks to human health.

[0013] There therefore remains an obvious need to develop, in particular according to the principles of ecobiology, cosmetic compositions, non-pharmaceutical, in particular ecobiological, which make it possible to fight effectively and sustainably (i.e., in the long term) against skin aging and the appearance of its signs, and which make it possible to promote skin regeneration, in particular healing, especially of aged skin. DETAILED DESCRIPTION OF THE INVENTION

[0014] Surprisingly, the Applicant, continuing its ecobiological approach to action on the cause of skin disorders, found that a combination of compounds, safe and suitable for human cosmetic use, act in synergy to meet these needs.

[0015] More specifically, the Applicant has found that the combination of particular compounds (also referred to as active ingredients) allows for a synergistic increase in the expression of the "Heparin-Binding Epidermal Growth factor" receptor (i.e., HBEGF), a constituent of the hyalurosome (Bames et al. 2010) known to stimulate the proliferation of keratinocytes (Kaya et al., 2006) and necessary for proper wound healing (Thônes et al., 2019).

[0016] Thus, a first object of the present invention relates to a cosmetic composition, advantageously eco-biological, comprising:

[0017] -at least one bacterioruberin;

[0018] -at least one active form of proline selected from the group consisting of dipalmitoyl hydroxyproline, hydroxyproline, acetyl proline, amidinoproline, cocoyl proline, dihydroxypropyl proline, proline, N-acetylhydroxyproline, and their salts.

[0019] The present invention offers various advantages, including preventing and / or effectively combating the signs of skin aging, in particular stimulating the growth and proliferation of fibroblasts, strengthening the dermo-epidermal barrier, combating oxidative stress, and promoting the production of structural proteins and glycans. Furthermore, the present invention promotes skin regeneration and repair, particularly in the context of dermatoporosis. The various effects of the com The benefits of the invention are quickly apparent and long-lasting. In other words, the invention is effective in the short, medium, and long term. Furthermore, regular application of the invention's composition, e.g., once or twice a day, enhances its beneficial effects.

[0020] In the context of the invention, the term "bacterioruberin" refers to a molecule of the bacterioruberin family (CAS No. 32719-43-0), which are C50 carotenoids characteristic of extremophile bacteria, particularly halophilic archaea and certain psychrophilic actinobacteria, and their derivatives, particularly glycosylated ones; bacterioruberins (CAS No. 32719-43-0) exist in various forms, the best known of which is α-bacterioruberin, which has the following structure:

[0021] [Chem.l]

[0022] In the context of the invention, the expressions "glycosylated bacterioruberin" or "glycosylated form of bacterioruberin" and "bacterioruberin in a glycosylated form" are used interchangeably and refer to a bacterioruberin in which at least one hydroxyl group is substituted with one or more, for example, 2 or 3, or even 4 sugar residues by means of an ether bond between the bacterioruberin backbone and the sugar. Bacterioruberins comprise 4 terminal hydroxyl groups, each of which is capable of being substituted by ether bond with a sugar-type group, or even one or more covalently linked sugars. For example, a "glycosylated bacterioruberin" as defined in the invention has the following structure:

[0023] [Chem.2]

[0024] In which R is independently chosen from a hydrogen atom and a sugar residue and where R at least one occurrence represents a sugar residue.

[0025] In practice, glycosylated or non-glycosylated bacterioruberins can be obtained by extraction and purification, for example by chromatography, of total carotenoid extracts from actinobacteria of the genera Micrococcus or Arthrobacter, advantageously the species A. agilis and / or M. roseus. The species A. agilis is also known as Micrococcus agilis. Thus, the extracts and strains described in Strand et al., 1997, Fong et al., 2001 and in document WO 2014 / 167247 can be used as a source of glycosylated bacterioruberins. Preferably, strains of A. The agilis used as sources of glycosylated bacterioruberins as defined in the invention are strain MB813 (described in Fong et al., 2001) and / or SB5 (described in document WO2014 / 167247). Methods for obtaining total carotenoid extracts from these bacterial species are known to those skilled in the art and are, for example, described in documents Strand et al., 1997, Fong et al., 2001 and WO2014 / 167247. .

[0026] In the context of the invention, the expressions "active form of proline" and "proline in active form" are used interchangeably and refer to proline (L-proline or D-proline, advantageously L-proline), which is an essential amino acid found in all organisms and animals and is a constituent of collagen; this particular amino acid is in an active form, that is to say, a form capable of exerting a biological action when this proline is administered to a subject, in particular a human.

[0027] In the context of the invention, "ecobiological" means respectful of the person, their interactions with the world, and the planet. The expression "ecobiological approach" refers to the specific approach initiated and developed by the inventor, which combines skin biology and ecology, particularly to help the skin function according to its natural biology over the long term.

[0028] In the context of the invention, the expression "signs of skin aging" refers to the appearance of wrinkles and fine lines, loss of firmness, density and / or volume of the skin, the appearance of pigment spots, loss of radiance of the skin, a change in the texture of the skin (it becomes rougher), sagging of the oval of the face, an uneven complexion, very visible pores and / or dry skin.

[0029] In the context of the invention, the terms "healing" and "to heal" are used interchangeably and refer to the various phases of the process by which tissue damage is repaired, or the integrity of damaged tissue is restored, following an injury. The different stages of this process are:

[0030] - an inflammatory phase: phagocytosis and elimination of bacteria and other impurities and the factors that produce the migration and division of cells that participate in the proliferative phase are released;

[0031] - a proliferative phase: angiogenesis, increased collagen content, formation of granular tissue, epithelialization and wound contraction; and

[0032] - a maturation and remodeling phase: the collagen is remodeled and realigned along the power lines and cells that are no longer needed are eliminated by apoptosis.

[0033] In the context of the invention, the term "active ingredient" refers to a compound possessing biological and / or therapeutic properties that underlie a physiological effect. The active ingredient is to be distinguished from an excipient present in the composition according to the invention.

[0034] In the context of the invention, the term "excipient" means a compound other than an active ingredient which gives the composition properties, in particular, of consistency, galenic and / or vectorization of the active ingredient.

[0035] In the context of the invention, the expressions "composition for cutaneous application" and "for topical use" are used interchangeably and refer to a composition compatible with application to the skin, mucous membranes, hair and / or scalp, preferably human skin.

[0036] In the context of the invention, the terms "blurring agent," "soft-focus particle," and "blurring particle" are used interchangeably and refer to a particle, advantageously colored, intended to give greater transparency to the complexion and a blurring effect. In particular, the blurring particle allows the composition containing it to attenuate, through an optical effect, the skin's microrelief, and in particular skin defects such as spots, wrinkles, and fine lines.

[0037] In the context of the invention, the terms "moisturizing compound", "moisturizing active ingredient", "moisturizing agent" and "moisturizer" are used interchangeably and refer to an active ingredient / compound capable of improving the skin's hydration status. This hydration status can be measured by means known to those skilled in the art, for example, by corneometric analysis.

[0038] Preferably, the present invention relates to a cosmetic composition, as defined above and having the following technical characteristics, taken alone or in combination:

[0039] - at least one active form of proline is selected in the group constituted by dipalmitoyl hydroxyproline, hydroxyproline and their salts;

[0040] - at least one active form of proline represents between 0.001% and 10% by weight total composition, advantageously between 0.01% and 5%, preferably between 0.1% and 2%;

[0041] - at least one bacterioruberin represents between 0.00001% and 5% by total weight of the composition, advantageously between 0.0001% and 1%;

[0042] - at least one bacterioruberin is derived from a bacterial extract;

[0043] - at least one bacterioruberin is present in glycosylated form;

[0044] - at least one bacterioruberin is chemically synthesized;

[0045] - the composition further comprises at least two bacterioruberins, avanta vastly different;

[0046] - one of the at least two bacterioruberins is in a glycosylated form;

[0047] - the at least two bacterioruberins represent between 0.00001% and 5% by weight total composition, advantageously between 0.0001% and 1%;

[0048] - the at least two bacterioruberins are derived from a bacterial extract;

[0049] - the at least two bacterioruberins are derived from the same bacterial extract;

[0050] - at least two bacterioruberins are chemically synthesized;

[0051] - the bacterial extract is an extract of Micrococcus agilis;

[0052] - the ratio between at least bacterioruberin and at least one active form of proline is between 1 / 10000 and 1 / 1;

[0053] - the ratio between the at least two bacterioruberins and the at least one active form of proline is between 1 / 10000 and 1 / 1;

[0054] - at least one bacterioruberin corresponds to the INCI designation Micrococcus lysate;

[0055] - the at least two bacterioruberins correspond to the INCI designation Mi crococcus lysate;

[0056] - at least one bacterioruberin is an α-bacterioruberin;

[0057] - at least one bacterioruberin corresponds to the INCI designation Halobacterium salinarum carotenoids;

[0058] - at least one bacterioruberin corresponds to the raw material HALORUBIN marketed by the company HALOTEK GMBH;

[0059] - the at least two bacterioruberins, glycosylated or not, correspond to the de INCI sign Micrococcus lysate;

[0060] - the at least two bacterioruberins, glycosylated or not, correspond to the de INCI signature CAPRYLIC / CAPRIC TRIGLYCERIDE & MICROCOCCUS LYSATE & TOCOPHEROL;

[0061] - at least one bacterioruberin corresponds to the raw material MIRO- RUBERINE marketed by the company GREENTECH;

[0062] - the at least two bacterioruberins, glycosylated or not, correspond to the matter first MIRORUBERINE marketed by the company GREENTECH;

[0063] - at least one active form of proline corresponds to the Oristar HPRO materials and Oristar PRO marketed by Orient Stars LLC and correspond respectively to the INCI designations hydroxyproline and proline;

[0064] - at least one active form of proline corresponds to the raw material Natisol marketed by the company Sinerga SPA and corresponds to the INCI designation cocoyl proline;

[0065] - at least one active form of proline corresponds to the raw material N- Acetyl-L-Proline marketed by Naris Cosmetics Co., LTD and corresponding to the INCI designation acetyl proline;

[0066] - at least one active form of proline is dipalmitoyl hydroxyproline (No CAS 41672-81-5) which is a lipophilic N-acylated derivative of the amino acid hydroxyproline, the purity of which is advantageously greater than 90%, even more advantageously greater than 95%, or even 98%, corresponding to the crystalline raw material DIPALMITOYL HYDROXYPROLINE marketed by the company SEPPIC under the name SEPILIFT DPHP™;

[0067] - the composition is in a form suitable for application by topical, particularly on the skin, mucous membranes, hair and / or scalp, preferably of a human subject;

[0068] - - the composition is ecobiological;

[0069] - the composition is in the form of an aqueous, hydroalcoholic solution, organic or oily; a suspension or dispersion in solvents or fatty substances, of a lotion type or a serum; in the form of a vesicular dispersion; in the form of a water-in-oil (W / O), oil-in-water (O / W) or multiple emulsion such as a water-in-oil-in-water (W / O / W) emulsion, the emulsion may be more or less thick and may be in the form of a cream or a milk; the composition of the invention may also be in the form of an ointment, a gel, a solid stick, anhydrous pasty or solid products, a foam, in particular an aerosol, a biphasic composition or a sprayable composition;

[0070] - the composition further comprises at least one particle, advantageously coloured, more advantageously with a Router effect;

[0071] - at least one particle represents between 0.5% and 20% by total weight of the com position, advantageously between 1% and 10%, preferably between 1% and 5%;

[0072] - at least one particle is chosen from the raw materials in the Ro- range nasphere (MERCK) based on a silica substrate, in particular RONASPHERE FLAWLESS, as well as several raw materials marketed by the companies JGC-C&C (Coverleaf range based on sericite or sericite / talc substrate), Nihon Koken (Relief Color range, Silséem based on mica / silica substrate), or Miyoshi Kasei (PC-Bail range based on silica substrate);

[0073] - at least one particle is a composite particle comprising sericite, brown iron oxide, titanium dioxide and silica. Such particles are marketed, for example, under the references COVERLEAF NS or JS or MF by the company CHEMICALS AND CATALYSTS;

[0074] - at least one particle has a structure that is of the silica microsphere type containing iron oxide, such as that marketed by the company MIYOSHI under the reference PC BALL PC-LL-100 P, this pigment being made up of silica microspheres containing yellow iron oxide;

[0075] - at least one particle corresponds to a composite particle having a structure of the sericite / brown iron oxide / titanium dioxide / silica type such as that marketed under the reference COVERLEAF MF by the company CHEMICALS AND CATALYSES. Alternatively, it may be cellulose derivatives, such as CELLULOBEADS D-5 marketed by the company DAITO KASEI;

[0076] - at least one particle corresponds to the raw material COVAMED PMMA 2 MUSI, marketed by SENSIENT COSMETICS TECHNOLOGY and corresponding to the INCI designations Methyl methacrylate crosspolymer and Silica; and / or

[0077] - at least one particle corresponds to the raw material KSP-100 commer specialized by the company SHINETSU corresponding to the INCI designation Vinyl di-methicone / methicone silsesquioxane crosspolymer;

[0078] - at least one particle corresponds to the commercially available raw material D-400 by the company TOCHI corresponding to the INCI designations HDI / trimethylol hexyllactone crosspolymer and Silica.

[0079] According to one particular aspect, the composition of the invention further comprises an aqueous base having a defined composition and containing, in particular: calcium chloride; potassium chloride; potassium phosphate; magnesium sulfate; sodium chloride; sodium bicarbonate; sodium phosphate; sodium citrate; citric acid; camosin; and hypotaurine (2-aminoethanesulfinic acid). Said aqueous base comprises:

[0080] - From 0.000001% to 0.1% by total weight of the aqueous calcium chloride base, advantageously from 0.0001% to 0.1%;

[0081] - From 0.0001% to 0.1% by total weight of the aqueous potassium chloride base 5, advantageously from 0.001% to 0.1%;

[0082] - From 0.00001% to 0.1% by total weight of the aqueous potassium phosphate base, advantageously from 0.0001% to 0.1%;

[0083] - From 0.0001% to 0.1% by total weight of the aqueous magnesium sulfate base, advantageously from 0.001% to 0.1%;

[0084] - From 0.001 to 4% by total weight of the aqueous sodium chloride base, avanta increasing from 0.1 to 4%;

[0085] - From 0.0001 to 0.1% by total weight of the aqueous hydrogen carbonate base of sodium;

[0086] - From 0.0001% to 0.1% by total weight of the aqueous hydrogen phosphate base of sodium;

[0087] - From 0.0001% to 1% by total weight of the aqueous citric acid base, advantageous slightly from 0.001% to 1%;

[0088] - From 0.001% to 1% by total weight of the aqueous sodium citrate base;

[0089] - From 0.0001% to 1% by total weight of the aqueous carnosine base, advantageously from 0.001% to 1%;

[0090] - From 0.0001 to 1% by total weight of the aqueous base of 2-aminoethanesulfinic acid, advantageously from 0.001 to 1%.

[0091] According to another particular aspect, the composition of the invention further comprises a bioactive system combining, on the one hand, a stable aqueous form of a nucleotide selected from ATP (adenosine triphosphate), Gp4G (diguanosine tetraphosphate), and Ap4A (diadenosine tetraphosphate); and on the other hand, at least one biomimetic peptide comprising at most six amino acids, mimicking a skin polypeptide or a skin protein, or a biomolecule that is an agonist or antagonist of said peptide or protein. In practice, the combination of these active ingredients makes it possible to catalyze the metabolic activity of skin cells while obtaining a dermocosmetic or therapeutic effect through the use of the biomimetic peptides. In practice, in the bioactive system according to the invention, the nucleotide represents at most 10% by weight of the composition, preferably between 0.001% and 5%; and the biomimetic peptide represents between 0.001% and 1% by weight of the composition..

[0092] According to another particular aspect, the composition of the invention further comprises peptide extracts of soy and / or wheat. In practice, the peptide extracts are obtained from soybeans and wheat by enzymatic hydrolysis of said seeds using peptidases, which allows the recovery of peptides with an average size of 700 Daltons. Preferably, the soybean peptide extract is the extract identified under CAS number 68607-88-5, and the wheat peptide extract is the extract identified under CAS number 70084-87-6. The wheat and soybean extracts may correspond to the INCI designations Hydrolyzed wheat protein and Hydrolyzed soy protein, respectively. In a particular embodiment, the peptide extracts of soy and wheat are used together, for example in a weight ratio of 80 / 20 and 20 / 80 respectively, advantageously between 70 / 30 and 30 / 70, preferably equal to 60 / 40.In an advantageous embodiment, the soy and / or wheat peptide extracts are free from synthetic tripeptides GHK (glycylhistidyllysine; INCI: Tripeptide-1). In practice, the soy and / or wheat peptide extracts represent between 0.01% and 20% by weight of the composition, advantageously between 0.1% and 10%, preferably between 0.2% and 0.7%.

[0093] The pharmaceutical form of the composition and its method of preparation, and consequently The excipients suitable for the composition of the invention can be chosen by a person skilled in the art based on their general knowledge, depending on the type of composition desired. In particular, the composition may include any fat commonly used in cosmetics. Examples include silicone-based fats such as silicone oils, gums, and waxes, as well as non-silicon-based fats such as oils and waxes of vegetable, mineral, animal, and / or synthetic origin. The oils may be volatile or non-volatile. Other examples include hydrocarbons, synthetic esters and ethers, fatty alcohols, and fatty acids.The composition may also include an aqueous medium, a hydroalcoholic medium containing an alcohol such as ethanol or isopropanol, or an organic medium comprising common organic solvents such as Cl-6 alcohols, including ethanol and isopropanol, glycols such as propylene glycol, and ketones.

[0094] The composition may include at least one conventional emulsifier, chosen from amphoteric, anionic, cationic or non-ionic emulsifiers, used alone or in mixture.

[0095] It may also include the usual adjuvants in the field concerned, such as hydrophilic or lipophilic thickeners or gelling agents, hydrophilic or lipophilic additives, active ingredients in particular cosmetics, preservatives, antioxidants, perfumes, fillers, pigments, UV filters, odor absorbers, colorants, moisturizers (glycerin), vitamins, essential fatty acids, liposoluble polymers in particular hydrocarbons, opacifiers, stabilizers, sequestrants, conditioners and propellants.

[0096] Of course, a person skilled in the art will take care to choose this or these possible additional adjuvants or excipients, and / or their quantity, in such a way that the advantageous properties of the composition according to the invention are not, or substantially not, altered by the envisaged addition.

[0097] It may be particularly advantageous to formulate the composition of the invention so that it is sprayable. This can be achieved, for example, by formulating specific emulsions comprising particular combinations of excipients.

[0098] Another object of the present invention also relates to the cosmetic use of the composition as described above, to prevent and / or treat the appearance of signs of skin aging, advantageously to prevent and / or combat skin aging and / or the appearance of wrinkles and fine lines and / or to induce the synthesis of constituent compounds of the extracellular matrix, advantageously proteglycans (PG), glycosaminoglycans (GAG) or collagen.

[0099] Another object of the present invention also relates to the cosmetic use of the composition as described above, to stimulate the growth and proliferation (i.e., hyperplasia) of fibroblasts, and to strengthen the dermo-epidermal junction, and / or combat oxidative stress.

[0100] Another object of the present invention also relates to the cosmetic use of the composition as described above, to prevent and / or treat dermatoporosis.

[0101] Another object of the present invention also relates to the cosmetic use of the composition as described above, to regenerate the skin, advantageously to promote skin healing, preferably via an acceleration of at least one of the phases of the healing process, advantageously the inflammatory phase and / or the proliferative phase and / or the maturation and remodeling phase.

[0102] Another object of the present invention also relates to the composition as described above for use to prevent and / or treat the appearance of signs of skin aging, advantageously to prevent and / or combat skin aging and / or the appearance of wrinkles and fine lines and / or to induce the synthesis of constituent compounds of the extracellular matrix, advantageously proteglycans (PG), glycosaminoglycans (GAG) or collagen.

[0103] Another object of the present invention also relates to the composition as described above for use in stimulating the growth and proliferation (i.e., hyperplasia) of fibroblasts, strengthening the dermo-epidermal junction, and / or combating oxidative stress.

[0104] Another object of the present invention also relates to the composition as described above for use in preventing and / or treating dermatoporosis.

[0105] Another object of the present invention also relates to the composition as described above for use in regenerating the skin, advantageously to promote skin healing, preferably via an acceleration of at least one of the phases of the healing process, advantageously the inflammatory phase and / or the proliferative phase and / or the maturation and remodeling phase.

[0106] Another object of the present invention relates to a cosmetic, non-therapeutic treatment method for the skin, in particular of a human subject, intended to prevent and / or treat the appearance of signs of skin aging, advantageously to prevent and / or combat skin aging and / or the appearance of wrinkles and fine lines and / or to induce the synthesis of constituent compounds of the extracellular matrix, advantageously proteglycans (PG), glycosaminoglycans (GAG) or collagen, to stimulate the growth and proliferation (i.e., hyperplasia) of fibroblasts, strengthen the dermo-epidermal junction, and / or combat oxidative stress.

[0107] Another object of the present invention relates to a cosmetic, non-therapeutic skin treatment method, in particular for a human subject, intended to prevent and / or treat dermatoporosis.

[0108] Another object of the present invention relates to a treatment method cosmetic, non-therapeutic, of the skin, particularly of a human subject, intended to regenerate the skin, advantageously promoting skin healing, preferably via an acceleration of at least one of the phases of the healing process, advantageously the inflammatory phase and / or the proliferative phase and / or the maturation and remodeling phase.

[0109] Any one of the cosmetic, non-therapeutic skin treatment processes described above includes a step of applying the composition of the invention to the skin at least once, advantageously twice, per day for a period of at least one month, advantageously two months, or even three, and even more advantageously six months.

[0110] Another object of the present invention relates to a kit comprising a moisturizing active compound and a composition according to the invention.

[0111] In particular, the moisturizing compound and the composition of the invention are in the form of a combined preparation intended for simultaneous, separate, or sequential use for: - To prevent and / or treat the appearance of signs of skin aging, advantageously to prevent and / or combat skin aging and / or the appearance of wrinkles and fine lines and / or to induce the synthesis of constituent compounds of the extracellular matrix, advantageously proteglycans (PG), glycosaminoglycans (GAG) or collagen, to stimulate the growth and proliferation (i.e., hyperplasia) of fibroblasts, strengthen the dermo-epidermal junction, and / or combat oxidative stress; - To prevent and / or treat dermatoporosis; and / or - To regenerate the skin, advantageously to promote skin healing, preferably via an acceleration of at least one of the phases of the healing process, advantageously the inflammatory phase and / or the proliferative phase and / or the maturation and remodeling phase.

[0112] The manner in which the invention can be implemented and the resulting advantages will be more apparent from the following embodiment examples, given by way of illustration and not limitation, in support of the attached figure. Figures

[0113] [Fig. 1]: Evaluation of the synergistic effect of the composition according to the invention on the expression of the HBEGF gene in human dermal fibroblasts. The results are from an experiment with 3 replicates per condition. Statistics: t-test, unpaired, two-tailed; *: 0.05 < p-value < 0.01.

[0114] EXAMPLES OF IMPLEMENTATION

[0115] The percentages indicated are given as a percentage of product weight relative to the total weight of the composition in the tables below.

[0116] Example 1: Cosmetic composition according to the invention - Facial day serum

[0117] A composition according to the invention is described in Table 1.

[0118] [Tables 1] AW / Af 7% Aqua / ww / eatt 52^38 Dicaprylyl carbonate 28.0873 Glycerin 5» Inbehœm PSCMO esters 3.œ f sred^d 2.00 CKMS triglycerides 23X1 Msthylpropanedi ûl 1.40 Dipahatiwl 1.00 PeîHaerythrilyitetra-dM^ Sugar" steàmte MW Hydmxy^thylacry^^ aenieykiimethyl taurate 0.88 connlvmer Puisvi eue d wol V K.1 0.50 Taœpheryl acetate 0.50 Sodium citrate 0.20 L2-hexan«dù?[ OJ25 t apsdri cl 0.125 Sodi um aœtyl atedhya 1 u ronate 040 Xylose 0.10 Citric acid 0.09 Polywb» 60 0.00 Sorbifau istMeàr^te 0.06 Adenosme 0.05 Rhamoose 0.05 M kreoteens hsate 0.01 Cellulose 0.02 nwiphejoî 0.0027 Fmctœügosacchartdes OJKH Capryiic / capric triglyceride 0.00095 Lrwuww fteWtoa «ex tract 0.00005

[0119] Example 2: Cosmetic composition within the meaning of the invention - Body cream

[0120] A composition according to the invention is described in Table 2.

[0121] [Tables2] Ota mine G / 5(7 AqtuÀvaM 'eau * u.imw Gh cei in y' tiigl^cnde 'WW h <q\medn ? 3.50 1000 CeM pahmtjtc 5.00OW l hturab* 23Wmo Si lies 2.0CHWÜ Mcohoi 1.921800 i Vivait y ï akohel I.2(W)0 BdKml a kohol umw G pnloU gvnv i.ooooœ GUu olive। fruit od 1.000000 pot<KMmn vciyl phosphate 1.000000 Ti mlem L0O0OO0 Zca । otsm sï tu h f tjs-utw Fragrance tp«a hum o.?{li lot > 0 Cetearvl shnoMde O.hiiRKïO Hdîanthus ànnaus fswdowâ seed «il 0.030000

[0122] Example 3: Study of the effect of the active ingredients according to the invention on the expression of the HBFEG gene 1. Objective of the study

[0123] The objective of the study was to evaluate the effect of the assets according to the invention on the expression of the HBFEG gene. 2. Materials and methods

[0124] 2.1. Biological models

[0125] Normal human dermal fibroblasts (NHDF).

[0126] - Cell type: NHDF, Bioaltematives PF2 reference used in the 8th pass.

[0127] - Growing conditions: 37°C, 5% CO2.

[0128] - Culture medium: DMEM optimized for the test, supplemented with calf serum fetal (FCS) 10%.

[0129] - Test medium: DMEM optimized for the test, supplemented with 1% FCS.

[0130] 2.2. Tested active ingredients

[0131] The compounds / actives according to the invention tested are described in Table 3.

[0132] [Tables3] INCI Concentrations tested in DMSO: Micrococcus lysate 0.823 pg / L, Dipalmitoyl hydroxyproline (DPHP) 1.37 mg / L

[0133] The active ingredients were solubilized in a mixture of DMSO and ethanol, the concentration of which in the samples was 0.06% DMSO and 0.012% ethanol. The concentrations of the active ingredients to be tested were chosen based on the results of cell viability tests (MTT tests) and were non-cytotoxic under the analytical conditions.

[0134] 2.3. Gene expression profile in NHDFs

[0135] a) Culture and processing

[0136] Fibroblasts were seeded in 24-well plates and cultured for 24 hours in culture medium and then in test medium for an additional 24 hours. The medium was then replaced with test medium containing or not (control) the compounds, combinations, solvent control (DMSO + 0.06% + 0.012% ethanol), or reference (TGF-3 to 10 ng / ml), and the cells were incubated for 24 hours. All experimental conditions were performed at n=3. At the end of incubation, the cells were washed in phosphate-buffered saline (PBS) and immediately frozen at 80°C.

[0137] b) Differential gene expression analysis

[0138] Marker expression was analyzed by RT-qPCR on total RNA extracted from cell monolayers of each experimental condition. Transcript analysis was performed in n=2 using a PCR array ("MV220909 - mQPA 16 customized") dedicated to research and adapted to the 'screening' format (Marker qPCR array or 'mQPA') and also targeting the HBEGF gene.

[0139] c). RNA extraction and reverse transcription

[0140] Total RNA was extracted from each sample using the TriPure Isolation Reagent™ according to the manufacturer's instructions. RNA quality was assessed by capillary electrophoresis (Bioanalyzer 2100, Agilent Technologies). RNA quantity was assessed using a spectrophotometer (Synergy Hl, BioTek Instruments).

[0141] Complementary DNA (cDNA) was synthesized by reverse transcription of total RNA in the presence of oligo(dT) and "Transcriptor Reverse Transcriptase" (Roche). The cDNA quantities were then adjusted before the PCR step.

[0142] d) Quantitative PCR

[0143] The incorporation of fluorescence into the amplified DNA was continuously measured during the PCR cycles. This resulted in a plot of fluorescence intensity as a function of the PCR cycle, allowing the evaluation of a relative expression (RE) value for each marker. The "control" condition corresponds to the DMSO solvent control used to solubilize the actives (i.e., a 0.06% DMSO and 0.012% ethanol solution).

[0144] The value selected for calculating the ER is the "exit point" (Ct) of the fluorescence curve. For a given marker, the higher value is the number of cycles; the lower value is the amount of mRNA.

[0145] The value of RE was calculated using the formula: (l / 2 number of cycles) x 106.

[0146] The PCR template used in the present study included a reference gene (GAPDH). This reference gene was used for data normalization because its expression is constitutive and theoretically stable. Therefore, the expression level of the target markers was compared to the expression level of this marker for all test conditions. PCR (Polymerase Chain Reaction) was performed using the LightCycler™ system (Roche Molecular Systems Inc.) according to the manufacturer's instructions.

[0147] The reaction mixture (final 10 pl) was prepared as follows:

[0148] - 2.5 pl of cDNA,

[0149] - primers (front and rear),

[0150] - mixture of reagents (Ozyme) containing DNA polymerase taq, SYBR Green I and MgCl2.

[0151] e) Data analysis and processing

[0152] The data were analyzed and processed using Microsoft Excel. Significance was determined using a two-tailed, unpaired t-test. Differences are considered statistically significant from p<0.05. (NS: p>0.05; *: p<0.05; **: p<0.01; ***: p<0.001). 3. Results and conclusions

[0153] The data are represented by [Fig. 1].

[0154] The synergy between two active substances (in this case: bacterioruberins (i.e., Micrococcus lysate) and an active form of proline (i.e., DPHP)) is calculated by comparing the results of tests in which the efficacy of the two substances and that of their mixture is measured. The Colby formula (Tammes, 1964; Schuldt, PH and Lammont 1962) is then used.

[0155] If the measured efficiency of the mixture is greater than that expected according to the Colby formula, there is synergy. If it is less, there is antagonism. And if it is When equal, the two substances act without interacting with each other.

[0156] Colby formula: E = X+Y - (XxY) / 100

[0157] With X = effect of the active ingredients Micrococcus lysate on the expression of the gene encoding HBEGF at the tested dose (0.823 pg / L), i.e., 120% relative expression of the HBEGF gene

[0158] With Y = effect of the active ingredient DPHP on the expression of the gene encoding HBEGF at the tested dose (1.37 mg / L), i.e., 130% relative expression of the HBEGF gene

[0159] With E = expected efficacy of the active ingredients Micrococcus lysate and DPHP at respective doses of 0.823 pg / L and 1.37 mg / L, without synergy = (120+130) - [(120*130) / 100] = 94%.

[0160] The observed effectiveness of the composition is 180%, i.e. well above 94% which was the expected effectiveness of the composition.

[0161] These data demonstrate the synergistic effect between the ingredients Micrococcus lysate and DPHP in a composition according to the invention.

[0162] At the tested concentration, the active ingredient Micrococcus lysate, which is a bacterial extract containing bacterioruberins in glycosylated or non-glycosylated form, has no significant positive effect on HBEGF gene expression. The active ingredient DPHP (dipalmitoyl hydroxyproline) induces HBEGF gene expression significantly compared to the control. The combination of the active ingredients Micrococcus lysate and DPHP has a significant synergistic effect on HBEGF gene expression.

[0163] BIBLIOGRAPHY

[0164] Barnes L, Tran C, Sorg O, Hotz R, Grand D, Carraux P, Didierjean L, Stamenkovic I, Saurat JH, Kaya G. Synergistic effect of hyaluronate fragments in retinaldehyde-induced skin hyperplasia which is a Cd44-dependent phenomenon. PLoS One. 2010 Dec 16;5(12):el4372.

[0165] Fong N, Burgess M, Barrow K, Glenn D (2001) Carotenoid accumulation in the psy-chrotrophic bacterium Arthrobacter agilis in response to thermal and sait stress Appl Microbiol Biotechnol 56: 750-756.

[0166] Kaya G, Tran C, Sorg O, Hotz R, Grand D, Carraux P, Didierjean L, Stamenkovic I, Saurat JH. (2006) Hyaluronate fragments reverse skin atrophy by a CD44-dependent mechanism. PLoS Med. 3:e493.

[0167] Mei N, Xia Q, Chen L, Moore MM, Fu PP, Chen T. (2005) Photomutagenicity of retinyl palmitate by ultraviolet A irradiation in mouse lymphoma cells. Toxicol Sci 88:142-149.

[0168] Strand A, Shivaji, S, Liaaen-Jensen (1997) Bacterial carotenoids 55. C50-carotenoids 25: revised structures of carotenoids associated with membranes in psychrotrophic Micrococcus roseus. Bioch. Syst. & Eco. 25: 547-552.

[0169] Thônes S, Rother S, Wippold T, Blaszkiewicz J, Balamurugan K, Moeller S, Ruiz- Gômez G, Schnabelrauch M, Scharnweber D, Saalbach A, Rademann J, Pisabarro MT, Hintze V, Anderegg U. (2019) Hyaluronan / collagen hydrogels containing sulfated hyaluronan improve wound healing by sustained release of heparin-binding EGF-like growth factor. Acta Biomater. 1 :135-147.

Claims

Demands

1. Cosmetic composition comprising: -at least one bacterioruberin; -at least one active form of proline selected from the group consisting of dipalmitoyl hydroxyproline, hydroxyproline, acetyl proline, amidinoproline, cocoyl proline, dihydroxypropyl proline, proline, N-acetylhydroxyproline and their salts.

2. Composition according to claim 1, characterized in that at least one active form of proline, advantageously dipalmitoyl hydroxyproline, hydroxyproline or one of their salts, represents between 0.001% and 10% by total weight of the composition, advantageously between 0.01% and 5%, preferably between 0.1% and 2%.

3. Composition according to any one of the preceding claims, characterized in that it comprises at least two bacterioruberins, advantageously different, more advantageously one of the at least two bacterioruberins being in a glycosylated form.

4. Composition according to any one of the preceding claims, characterized in that at least one bacterioruberin or at least two bacterioruberins represent between 0.00001% and 5% by total weight of the composition, advantageously between 0.0001% and 1%.

5. Composition according to any one of the preceding claims, characterized in that at least one bacterioruberin or at least two bacterioruberins are derived from a bacterial extract, advantageously from the same bacterial extract.

6. Composition according to claim 5, characterized in that the bacterial extract is an extract of Micrococcus agilis.

7. Composition according to any one of the preceding claims, characterized in that at least one bacterioruberin or at least two bacterioruberins correspond to the INCI designation Micrococcus lysate.

8. Composition according to any one of the preceding claims, characterized in that at least one bacterioruberin or at least two bacterioruberins correspond to the INCI designation Micrococcus lysate; and in that at least one active form of proline corresponds to the INCI designation Dipalmitoyl hydroxyproline.

9. Composition according to any one of the preceding claims for its use in a method for regenerating skin, advantage- gently to promote skin healing.

10. 10. Non-therapeutic and cosmetic skin treatment method, particularly human, intended to prevent and / or treat the skin signs of aging, in particular wrinkles, comprising at least the application to the skin of a cosmetic composition according to claim 1 to 8.

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