Submicron particle comprising a peptide active ingredient, preparation process and uses, particularly in cosmetics

Abstract

The submicron particle is substantially spherical, comprising an oily core containing the peptide active ingredient and a wax shell that is solid at room temperature. The particle is stabilized by an outer layer of a high HLB non-ionic surfactant. The invention proposes a process for obtaining an aqueous suspension of said particles with a satisfactory encapsulation rate and improved long-term stability, particularly for use in the cosmetics industry. Combinations of peptides can be integrated inside the particles and adsorbed on the outside. [Fig. 1]

Description

Title of the invention: Submicron particle comprising a peptide active ingredient, preparation process and uses, particularly cosmetics Field of invention

[0001] The subject of the present invention is a submicron particle comprising a peptide active ingredient, in particular a peptide or mixture of peptides from the field of cosmetics, a process for preparing said particle, and uses in particular for a non-therapeutic cosmetic treatment of the skin (including the scalp) and / or its appendages (such as hair, eyelashes, eyebrows, nails, hair) of mammals, humans or animals, and for manufacturing a topical or oral composition in particular intended for this treatment.

[0002] It mainly concerns the field of cosmetic products and hygiene and personal care products, applied topically.

[0003] By peptide active ingredient, according to the invention, is meant a peptide or mixture of peptides, said peptide comprising at least two amino acids, linked by an amide bond between one of their acid and amine functions. The peptide may be of synthetic or natural origin, obtained by extraction or by biotechnological means. It may be linear, cyclic or branched. By extension, the term peptide active also includes, according to the invention, an amino acid or mixture of free amino acids.

[0004] According to the invention, the term amino acid or amino acid means a natural or synthetic amino acid or one obtained by biotechnological means, of L, D or racemic stereochemistry. It may be chosen from natural amino acids, their derivatives (for example hydroxyproline) or analogues (for example 5-hydroxyproline or methionine sulfone or sulfoxide). The acid and amine functions may be separated by one or more carbons, one carbon corresponding to natural alpha-amino acids, two carbons corresponding to beta-amino acids (for example beta-alanine), three carbons to gamma-amino acids (for example GABA or gamma-aminobutyric acid), four carbons corresponding to delta-amino acids (for example 5-aminovaleric acid), etc. Prior art

[0005] Peptides, alone or in mixtures, are among the most popular biologically active ingredients in a topical cosmetic formula. Often consisting of fragments of amino acid sequences of proteins present in the skin, or derivatives of these fragments, they act in situ by biomimicry like these proteins.

[0006] Historical peptides, such as Matrixyl® or Matrixyl®3000 (from Sederma), have been developed for anti-aging applications thanks in particular to stimulation of the main proteins of the dermal extracellular matrix, collagen 1 and elastin. Today, numerous peptides are available, capable of acting on more varied targets and leading to highly anticipated cosmetic benefits such as hydration, slimming, sensory comfort of the skin, radiance of the complexion or its pigmentation, or even pigmentation of hair and body hair.

[0007] A major advance in the field of cosmetic peptides has been to derivatize the peptide to modify its lipophilic character and increase its bioavailability in the skin. The derivatization can be present in the C-terminal and / or N-terminal position of the peptide, formed for example by acylation, more particularly to attach a fatty chain to the amino acid sequence, in particular a C14 to C20 chain close to the chain lengths of skin lipids, preferably a C16 palmitoyl (Pal) or C14 myristoyl (Myr) or C18 oleoyl carbon chain. The Pal chain in the N-terminal position is often preferred because it does not oxidize and its biological activity has been proven.

[0008] Other types of derivatization have been proposed, such as the attachment of a Biotinoyl part in place of the fatty chain, or derivatives of particular acids such as those of ascorbic, retinoic, cinnamic, oleanolic, hyaluronic, ni-cotinic, lipoic, gallic or pantothenic acid.

[0009] Amino acids, derived in this way, are also used as active compounds in cosmetics, for example Oleyl-Tyrosine as a lightening compound, or Pal-Alanine, Pal-Glycine and Pal-Isoleucine as anti-aging active ingredients (Seppic Company).

[0010] The present invention relates more particularly to this type of peptide active agent having a derivatization in the N-terminal and / or C-terminal position, in particular a derivatization of a lipophilic nature, such as an acylation with a palmitoyl chain.

[0011] Peptide active ingredients can be marketed in powder form (particularly acetate or hydrochloride salts) with a high degree of purity.

[0012] Most often, in the cosmetics industry, peptide active ingredients are marketed in the form of an active ingredient, that is to say in the form of a pre-formula to be integrated directly into the final formula of the cosmetic composition.

[0013] This pre-formula corresponds to the peptide active ingredient solubilized in a solvent appropriate to the physicochemical characteristics of the peptide(s) and / or the amino acid(s) of the peptide active ingredient, such as glycerin, at a pH allowing the stability of the active ingredient to be maintained in this pre-formula.

[0014] One of the problems encountered is the stability over time of the ingredient constituted by the pre-formula. It is not possible to store the ingredient at room temperature. room temperature, but generally necessary to keep it at 4 / 5°C. Room temperature means, according to the invention, a temperature ranging from 15 to 30°C. Subject of the invention

[0015] The present invention aims to overcome this drawback, namely to offer a solution for stabilizing amino acids or peptides, in particular lipophilic by chemical nature or by derivatization, so that they can be easily stored and handled at room temperature to produce cosmetic formulations, without risk of degradation and loss of activity.

[0016] The present invention also aims to propose a means of improving the stability of the peptide active ingredient so that it retains optimal biological activity up to its target in the skin or its appendages, while being protected from oxidative and protease attacks.

[0017] It also aims to propose a solution which uses raw materials which are environmentally friendly, in particular by banning ethoxylated surfactants.

[0018] To this end, it proposes, according to a first object, a submicronic particle comprising a peptide active agent consisting of a peptide or mixture of peptides and / or an amino acid or mixture of amino acids, characterized in that it is substantially spherical consisting of an oily core containing said peptide active agent surrounded by a wax shell which is solid at room temperature, at least one non-ionic surfactant having an HLB substantially greater than or equal to 10 forming an outer layer on said wax shell.

[0019] The present invention proposes, according to a second object, a method for preparing particles according to the first object. To this end, it proposes a method for encapsulating a peptide active ingredient, comprising the following successive steps:

[0020] - a step of providing a mixture comprising:

[0021] - an aqueous phase in which the peptide active ingredient is solubilized optionally- ally thanks to a water-miscible co-solvent and / or a pH adjuster,

[0022] - a waxy phase solid at room temperature consisting essentially of a or more waxes; and

[0023] - an oily phase that is liquid at room temperature and essentially consists of a or several oils,

[0024] the mixture further comprising at least one non-ionic surfactant (TA1) having an HLB substantially greater than or equal to 10, and each phase being made homogeneous and liquid where appropriate by heating and / or by mechanical and / or physical means; and

[0025] - a step of hot formation of an emulsion of said mixture;

[0026] - a step of reducing the size of the particles of said emulsion substantially to a submicron size; and

[0027] - a cooling step so as to form a suspension of particles li submicron structured peptides in which said peptide active ingredient is encapsulated.

[0028] The submicron particles according to the invention comprise two types of lipids: lipids which are solid at room temperature and lipids which are liquid at room temperature. The particles are of the SLC type for "solid liquid nanocarrier". The mixture of these two types of lipids leads, according to the invention, to particles which are solid at room temperature thanks to a substantially waxy, therefore solid, envelope and an oily core in which the peptide active ingredient is trapped, protected from the outside by the waxy envelope. The term "structured" is used to characterize particles which are solid at room temperature (visible by Differential Scanning Calorimetry (DSC) analysis, see the detailed description below showing, for example, a melting temperature significantly higher than 30°C for particles encapsulating the peptide Pal-KTTKS).

[0029] The term "submicron particles" is used according to the invention for particles with a submicron average diameter, i.e. less than 1 pm (=1000 nm), and not the term "nanoparticles" because the particles according to the invention do not correspond to the definition of the WTO and the European Union regulations which limit the term "nano-material" to insoluble particles with a nominal diameter ranging from 1 to 100 nm. The term "average" submicron diameter corresponds to the average diameter of the particles at the 50th percentile of the distribution curve (D50).

[0030] Test results given later in the description show the structured lipid particles formed according to the method of the invention with a set of representative cosmetic peptides, their encapsulation rate, and their stability over time at room temperature.

[0031] The results show in particular:

[0032] - that we obtain structured, spherical submicron particles and homogeneous at room temperature, without aggregation and phase shift during manufacture, following a Gaussian size profile around an average value of 100 to 900 nm, with low polydispersity (less than or equal to 2), a high encapsulation rate, and which may include a content after encapsulation compatible with use for formulating cosmetic products (up to 5000 ppm);

[0033] - that the particles are stable, without aggregation and dephasing, release of peptide compound(s) at room temperature for several months, and up to 1 year for Pal-KTTKS;

[0034] - that the particles are also stable, without aggregation and dephasing, release of peptide compound(s) at 40°C, stimulating accelerated stability;

[0035] - a slow release profile of the peptide compound(s), with a half-life suitable for topical diffusion, i.e. the stability of the peptide active ingredient is maintained during the period of release into the skin;

[0036] - from a formulation point of view, the peptide active being encapsulated and protected in a particle with a waxy envelope, it will be less sensitive to electrolytes and the risk of undesirable precipitation in the presence of certain adjuvants, such as carbomers, will be limited or even eliminated;

[0037] - that environmentally friendly raw materials can be put into work, in particular non-ethoxylated surfactants, such as lauryl glucoside or decyl glucoside.

[0038] An additional advantage according to the invention lies in the fact that the penetration of the peptide compounds through the stratum corneum will be promoted and even into deeper layers of the skin, the lipid composition of the particles and their very small size (submicronic) in fact contributing to increasing the adhesion of the particles and their contact surface with the stratum corneum thanks to a lipid rearrangement.

[0039] Yet another advantage of the process according to the invention lies in the fact that industrialization is simple, organic solvents can be avoided and production times are short. High-pressure homogenizers are commonly used in many industries, particularly the cosmetics industry, to obtain submicron particles.

[0040] According to the invention, the wax(es) are chosen so as to be solid at room temperature, that is to say having a melting point alone or as a mixture above 30°C, preferably from 40°C to 80°C, more preferably from 42°C to 65°C.

[0041] Preferably, the wax used is natural or synthetic, consisting of at least one ester of C12-C36 fatty acid(s) and C12-C36 fatty alcohol(s), preferably chosen from triglycerides, diglycerides, monoglycerides, and monoesters of C18-C36 fatty acid(s) and C18-C36 fatty alcohol(s), more preferably cetyl palmitate, glyceryl tribehenate, glyceryl stearate or tristearate, a wax of vegetable origin, such as rice wax, or a wax of animal origin, such as beeswax.

[0042] According to the method, this waxy phase is heated to the melting temperature of the wax or wax mixture and kept liquid at approximately this temperature for the steps of forming the suspension and reducing the size of the particles of the suspension.

[0043] Preferably, according to the invention, an oil or a mixture of oils is used, which by definition are liquid at room temperature, comprising:

[0044] - fatty acids essentially C8 to C18,

[0045] - mono-, di- or triesters of fatty acid(s) essentially C8 to C18, comprising mono-, di- or triglycerides of fatty acids; and / or

[0046] - fatty alcohols comprising C8 to C18 carbon chains.

[0047] Preferably, the oils are chosen from C8 to C18 fatty acid esters.

[0048] Preferably, the oils used according to the invention are of plant origin, in particular from soybean, rapeseed, sunflower, palm and / or coconut.

[0049] Preferably, in the process of the invention the oily phase is heated to the same temperature as the waxy phase. Also preferably, the waxy phase and the oily phase are heated simultaneously.

[0050] According to the method of the invention, the peptide active ingredient can be added pure in powder form directly into the aqueous phase or pre-solubilized in a fraction of the aqueous phase. If this peptide active ingredient is not soluble or is only partially soluble in the aqueous phase, one or more water-miscible co-solvents can be used in which the peptide compound or the mixture of peptide compounds is soluble, such as glycerin or sorbitol.

[0051] According to the method of the invention, it may be necessary in addition, with or without co-solvent(s), to aid the solubilization of the peptide compound, to add a fraction of the oil or mixture of oils, to apply a mechanical means, such as stirring, or other (microwaves, ultrasound, electromagnetic and / or electric waves), and / or to heat. It may also be necessary to add to the aqueous phase a pH adjuster, such as citric acid or NaOH, to acidify or alkalinize the medium and aid solubilization.

[0052] According to the invention, it may also be advantageous to use a second surfactant (TA2) having a low HLB, substantially less than or equal to 6. This second surfactant will promote the stabilization of the particles (size more stable over time and at a temperature above 40°C). According to preferred characteristics, this second surfactant is used. The particles according to the invention preferably comprise this second surfactant.

[0053] According to other characteristics of the invention, preferred and advantageous of the particles and / or of the process: (the % by weight being expressed relative to the total weight of the mixture or of the suspension)

[0054] - the % by weight of the wax or mixture of waxes is between 1 and 15, preferably between 1 and 10, preferably between 2 and 8; and / or

[0055] - the % by weight of the oil or mixture of oils is between 2 and 20, preferably between 5 and 15, more preferably between 7.5 and 12.5; and / or

[0056] - the ratio of % by weight of wax to % by weight of oil in the particles is between 1:1 and 1:10, preferably between 1:2 and 1:8, preferably between 1:2 and 1:5; and / or

[0057] - the % by weight of water is between 30 and 90, preferably between 45 and 90, of preferably still between 50 and 85; and / or

[0058] - the first surfactant TA1 preferably has an HLB greater than or equal to 12; and / or

[0059] - the second surfactant TA2, when present, preferably has an HLB between 3.5 and 5.5; and / or

[0060] - the first surfactant TA1 preferably comprises at least one alkylglucoside or an alkylpolyglucoside of at least one saturated C10-C16 fatty alcohol, and / or at least one polyglycerol ester of a saturated C10-C16 fatty acid, and / or at least one polysorbate (esters of etoxylated sorbitans), a poloxamer (formed from block chains of ethylene oxide and propylene oxide), and / or a polyvinyl derivative (consisting of vinyl acetate and vinylpyrrolidone, such as polyvinyl alcohol PVA or polyvinylpyrrolidone PVP); and / or

[0061] - preferably at least one alkylglucoside or an alkylpolyglucoside of at least one saturated C10-C16 fatty alcohol is chosen from lauryl glucoside, decyl glucoside, coco glucoside and their polyglucosidated derivatives, more preferably chosen from lauryl glucoside and decyl glucoside, and / or

[0062] - preferably at least one polyglycerol ester of saturated C10-C16 fatty acid is selected from polyglyceryl-4 to polyglyceryl-6 of lauric, sebacic, caprylic or capric acid, or a mixture thereof, preferably the mixture of polyglyceryl-4 laurate / sebacate and polyglyceryl-6 caprylate / caprate, or the mixture of polyglyceryl-4 laurate / sebacate and polyglyceryl-4 caprylate / caprate; and / or

[0063] - preferably the polysorbate is chosen from polysorbate 60 and polysorbate 80; and / or

[0064] - preferably the poloxamer is poloxamer 188;

[0065] - preferably the polyvinyl derivative is polyvinyl alcohol and / or

[0066] - the first surfactant TA1 is added to the aqueous phase, solubilized if necessary by heating; and / or

[0067] - the second surfactant TA2 is chosen from a phospholipid or a monoester in particular sorbitan, or a mixture of the two, the phospholipid being preferably chosen from phosphatidic acid, a phosphatidylcholine or lysophosphati-dylcholine, a glycerophosphocholine, a phosphatidylserine or a lysophosphati-dylserine, a phosphatidylethanolamine, a phosphatidylinositol or a sphin-gomyelin, more preferably the second surfactant being hydrogenated, and more preferably it is hydrogenated lecithin, the second surfactant TA2 being a phospholipid, it is preferably added to the waxy phase, solubilized if necessary by heating, the second surfactant TA2 being a sorbitan monoester, it is preferably chosen from sorbitan monostearate, sesquistearate sorbitan, the sorbitan laurate, sorbitan oleate, sorbitan sesquioleate and sorbitan isostearate, more preferably sorbitan isostearate, preferably added to the wax phase, solubilized by heating; and / or

[0068] - a wax is used which is natural or synthetic, preferably consisting of at least at least one ester of C12-C36 fatty acid(s) and C12-C36 fatty alcohol(s), more preferably chosen from triglycerides, diglycerides, monoglycerides, and monoesters of C18-C36 fatty acid(s) and C18-C36 fatty alcohol(s), more preferably cetyl palmitate, glyceryl tribehenate, glyceryl stearate or tristearate, a wax of vegetable origin, such as rice wax, or a wax of animal origin, such as beeswax; and / or

[0069] - an oil is used which is preferably formed from esters of essential fatty acid(s) primarily C16-C18, including monoesters such as ethyl oleate, or triglycerides such as caprylic / capric triglyceride mixture; and / or

[0070] - the percentage by weight of the first surfactant TA1 is from 1% to 6%, preferably still 2 to 4%; and / or

[0071] - the percentage by weight of the second surfactant TA2 is from 0.1 to 5%, preferably still from 0.5 to 2%; and / or

[0072] - the particle size reduction step is carried out by homogenization under high pressure, by emulsification by membranes, by micro-fluids, by rotor-stator of the Ultraturrax type; and / or

[0073] - the method comprises the addition of one or more preservatives and / or one or more antioxidants, and / or bacteriostatic agent, preferably from 0.1 to 5%, in one of the phases or in an additional phase, preferably in the aqueous phase, such as 1,2-octanediol, propanol, citric acid, hexanediol, sodium benzoate; and / or

[0074] - the final pH of said suspension is preferably between 3 and 9, compatible with a formula for the skin and / or its appendages; it varies depending on the peptide active ingredient; and / or

[0075] - when a co-solvent is present, its quantity is at most 30%, said co solvent found at the end of the process mixed with the aqueous phase.

[0076] Optionally, the method comprises an additional drying step where the water is removed so as to obtain a dried suspension, this step being able to be carried out by drying the aqueous phase to obtain a powder, for example by spray drying or freeze-drying, using in particular a sugar such as mannitol or maltodextrin as a drying aid.

[0077] Preferably, the particles according to the invention are in aqueous suspension, in which there may be, if necessary, a co-solvent used to help solubilize the peptide.

[0078] A particularly interesting combination of the process according to the invention is characterized in that the mixture comprises or consists of:

[0079] - 1 to 5000 ppm of peptide active;

[0080] - 2 to 8% by weight of a wax or a mixture of waxes;

[0081] - 7.5 to 12.5% ​​by weight of an oil or a mixture of oils;

[0082] - 2 to 4% by weight of lauryl glucoside or decyl glucoside;

[0083] - optionally from 0.5 to 2% by weight of phosphatidylcholine or phosphatidylserine hydrogenated; and

[0084] - optionally a co-solvent, a pH adjuster and / or a preservative and / or a bacteriostatic and / or antioxidant agent;

[0085] all percentages by weight being expressed as a function of the total weight of the mixture or suspension, and the complement qsp to 100% by weight being provided by the weight of water.

[0086] Optionally, the method according to the invention comprises a step of adding an additional peptide active ingredient to the suspension obtained after cooling and obtaining the solid particles. The additional peptide active ingredient is previously solubilized in water with, if necessary, a water-miscible co-solvent, then added and mixed with the suspension. This step advantageously makes it possible to adsorb a second peptide active ingredient outside the particles to transport it jointly to an active site.

[0087] Preferably, the free amino acid(s) and / or the peptide(s) constituting the peptide active agent comprise a derivatization, preferably in the N-terminal or C-terminal position, the peptide active agent corresponding to the following general formula (A): X-(Xaa)nZ

[0088] In which:

[0089] - Xaa is an amino acid, derivative or analogue;

[0090] - n an integer > 1, when n > 2 the amino acids being chosen independently depending on each other;

[0091] - at the N-terminal end X chosen from H, -CO-R1, -SO2-R1 or a group biotinoyl;

[0092] - at the C-terminal end Z chosen from OH, OR1, NH2, NHR1 or NR'R2; and

[0093] - R1 and R2 being, independently of one another, chosen from an alkyl group, aryl, aralkyl, alkylaryl, alkoxy, saccharide and aryloxy, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and / or sulfur-containing, said group having from 1 to 24 carbon atoms and which may have in its skeleton one or more heteroatoms O, S and / or N;

[0094] - the case where X=H and Z=OH being excluded.

[0095] Preferably according to the invention, the peptide active ingredient comprises one or more peptides. More preferably, the peptide(s) of the peptide active ingredient contain a sequence of at most 10 amino acids, which corresponds in formula (A) to 2 < n < 10, more preferably a sequence of at most 6 amino acids, which corresponds in formula (A) to 2 < n < 6.

[0096] According to the invention, the peptide active ingredient, when it is constituted by a mixture of peptides, preferably comprises a mixture of 2 to 5 peptides, more preferably a mixture of 2 peptides.

[0097] According to other preferred characteristics of the invention, in formula (A):

[0098] - R1 and / or R2 is an alkyl chain of 1 to 24 carbon atoms, preferably a lipophilic alkyl chain of 3 to 24 carbon atoms, more preferably of 8 to 24 carbon atoms, and more preferably of 12 to 16 carbon atoms; and / or

[0099] - X is an acyl group CO-R1; preferably chosen from an octanoyl (C8), decanoyl (C10), lauroyl (C12), myristoyl (C14), palmitoyl (C16), stearoyl (C18), biotinoyl, elaidoyl, oleoyl and lipoyl; more preferably selected from lauroyl (Cl2), myristoyl (Cl4) and palmitoyl (Cl6), and / or

[0100] - Z is chosen from OH, OMe, OEt and NH2, preferably OH; and / or

[0101] - X is chosen from palmitoyl (C16), myristoyl (C14), lauroyl (C12) or biotinoyl; more preferably palmitoyl (Cl6), and Z is OH.

[0102] Peptides comprising in the N or C terminal position derivatives of particular acids such as those of ascorbic, retinoic, cinnamic, oleanolic, hyaluronic, nicotinic, lipoic, gallic or pantothenic acid are also covered by the present invention.

[0103] Preferably, the amino acids of the peptide active ingredient and / or of the peptide(s) forming it are thus chosen from the amino acids (L and / or D): histidine (H, His), arginine (R, Arg), lysine (K, Lys), phenylalanine (F, Phe), alanine (A, Ala), leucine (L, Leu), methionine (M, Met), isoleucine (I, Ile), tryptophan (W, Trp), proline (P, Pro), valine (V, Val), cysteine ​​(C, Cys), glycine (G, Gly), glutamine (Q, Gin), asparagine (N, Asn), serine (S, Ser), tyrosine (Tyr, Y), threonine (T, Thr), aspartic acid (Asp, D) and glutamic acid (Glu, E), their oxygenated derivatives such as methionine sulfone or sulfoxide, or a hydroxyproline, and their structural analogues such as for example the lysine analogue omithine.

[0104] The peptide active ingredient according to the invention may be optically pure, or consist of: L or D isomers of amino acids or a mixture thereof. L isomers which are those present in the natural state may be preferred. It can be obtained in particular by synthetic or biotechnological means.

[0105] The peptide may be complexed with a metal ion (e.g. copper, zinc, manganese or magnesium).

[0106] The detailed description given below illustrates in detail all these advantages as well as others obtained thanks to the process of the invention. The di-, tri-, tetra-, penta-, hexa- The following representative cosmetic peptides are given as examples.

[0107] - The dipeptide Pal-PP (Palmitoyl Dipeptide-52) corresponding to the sequence palmitoylated proline-proline peptide. This peptide has propigmenting activity and can in particular prevent or treat canities. This peptide is, for example, marketed by the Applicant in solution under the trade name Sylverfree™.

[0108] - The tripeptide Pal-GHK (INCI name: Palmitoyl Tripeptide-1) corresponding to the palmitoylated glycine-histidine-lysine peptide sequence. This peptide stimulates the synthesis by fibroblasts of important molecules of the dermal extracellular matrix, such as collagen and elastin. It helps to strengthen the mechanical properties of the skin, elasticity and firmness, and therefore to treat wrinkles. This peptide is, for example, marketed in solution by the Applicant under the trade name Biopeptide CL™.

[0109] - Biot-GHK (INCI name: Biotinoyl Tripeptide-1) corresponds to the bio-version tinoylated Pal-GHK. It also helps stimulate the synthesis of collagen IV and laminin 5, and stimulate the activity of hair follicles. It is recommended in particular for preventing or treating hair loss. This peptide is, for example, marketed in solution by the Applicant under the trade name Procapil™.

[0110] - Pal-KMO2K (INCI name: Palmitoyl tripeptide-38) corresponds to the sequence palmitoylated lysine-methionine-lysine peptide, MO2 corresponding to a dioxygenated methionine. This peptide stimulates the production of collagens I and II, fibronectin and hyaluronic acid by dermal fibroblasts. It is recommended for anti-aging treatment, for example marketed in solution by the Applicant under the trade name Matrixyl®Synthe'6.

[0111] - Pal-GQPR (INCI name: Palmitoyl Tetrapeptide) (SEQ ID No. 1) corresponds to the palmitoylated glycine-glutamine-proline-arginine peptide sequence. This peptide acts on the excess production of interleukins. In particular, it helps to inhibit damage linked in particular to glycation. It is, for example, marketed by the Applicant in solution under the trade name Rigin™. It is also marketed in solution in a mixture with Pal-GHK by the Applicant under the trade name Matrixyl® 3000 for proven synergistic anti-aging activity. The encapsulation of the Pal-GHK / Pal-GQPR mixture is also given as an example later in the description.

[0112] - Pal-KTFK (INCI name: Palmitoyl Tetrapeptide-10) (SEQ ID No. 2) corresponds to the peptide sequence lysine-threonine-phenylalanine-lysine palmitoylated. This peptide has epidermal activity, preserving the skin barrier, smoothing the relief and stimulating in particular the alpha-crystalline chaperone protein for an effect on the transparency and radiance of the complexion. It is also described as stimulating the main dermal proteins. It is marketed in solution by the Applicant under the trade name Crystalide®.

[0113] - Pal-KTTKS (INCI Name: Palmitoyl pentapeptide-4) (SEQ ID No. 3) corresponds to the peptide sequence lysine-threonine-lysine-serine (KTTKS). This peptide has a very comprehensive activity on the dermis and epidermis. It is recommended as a global anti-aging product, particularly in solution under the trade name Matrixyl® by the Applicant.

[0114] - Pal-VGVAPG (INCI Name: Palmitoyl Hexapeptide-12) (SEQ ID No. 4) corresponds to the peptide sequence valine-glycine-valine-alanine-proline-pal-mitoylated glycine. This peptide stimulates the mobility of fibroblasts and acts on the suppleness of the skin. It is recommended to improve the firmness and tone of the skin. It is marketed in particular by the Applicant in solution in the commercial product Dermaxyl®.

[0115] The invention may be applied to peptides other than those mentioned above, in particular Pal-KTSKS (SEQ ID No. 5), Pal-K(P)HG (K(P) meaning a proline grafted onto a lysine), a cyclic peptide derived from flax seeds as described in WO2019 / 149150, Acetyl-Tyr-Arg-O-hexadecyl ester, Myr-PPL, Pal-LLAN (SEQ ID No. 6), Pal-YGGFL (SEQ ID No. 7), VW, Pal-K-Ava-K, Pal-Y(Me)-OH, Pal-KHG or Pal-KGH. Numerous examples of peptides for cosmetics are described in the Applicant's applications and patents to which the present invention may be applied. Amino acids or mixtures of amino acids may also be encapsulated according to the invention, in particular those mentioned above.

[0116] [Fig. 1] schematically illustrates the predictive structure of the particles obtained: a wax envelope surrounded by the first surfactant which stabilizes it, an oily core in which the peptide active ingredient is trapped, the second surfactant, when present, being found in the core of the particle and at the interface with the wax.

[0117] The present invention also has as a third subject an aqueous suspension of submicron structured lipid particles according to the first subject defined above, capable of being obtained by the process according to the invention, as defined according to the second subject defined above.

[0118] According to preferred and advantageous characteristics of the suspension according to the invention:

[0119] - the % by weight of peptide compound or mixture of peptide compounds encapsulated relative to the % by weight of said suspension is from 0.001 to 0.5%, preferably from 0.05 to 0.2%; more preferably from 0.01 to 0.1%; and / or

[0120] - the size of all the particles is from 0.1 to 1 pm, preferably from 0.15 to 0.6 pm, more preferably from 0.2 to 0.5 pm; and / or

[0121] - the polydispersity rate is from 0.1 to 2.0, preferably from 0.1 to 1.5, with a size average majority of particles (D50) between 0.2 and 0.9 pm, preferably between 0.2 and 0.4; and / or

[0122] - the particles are structured substantially spherical and have an envelope waxy; in addition, they comprise an oily core containing said peptide active ingredient; and / or

[0123] - the suspension is in a dried form (a powder), an advantageous form to formulate makeup products, for example.

[0124] An aqueous suspension according to the invention is water-dispersible and can be used to form cosmetic formulations of the emulsion, gel, serum, suspension type, for finished products for the skin and its appendages, such as hair products (shampoo, conditioner).

[0125] According to a fourth object, the present invention provides a composition, in particular a cosmetic composition for the skin and / or its appendages such as the hair, but which may also be a pharmaceutical composition, comprising as active ingredient an effective quantity of particles as defined according to the first object, or of a suspension, as defined above according to the third object, and a physiologically acceptable vehicle. Such a composition is preferably topical.

[0126] “Physiologically acceptable” means that the compositions are suitable for a topical or transdermal use, in contact with the mucous membranes, nails, scalp, hair, fur and skin of mammals and more particularly humans, compositions which can be ingested or injected into the skin, without risk of toxicity, incompatibility, instability, allergic response, and others. This “physiologically acceptable medium” forms what is classically called the excipient of the composition.

[0127] According to the invention, by “topical treatment” or “topical use” is meant an application which is intended to act at the place where it is applied: skin, mucous membrane and / or appendages.

[0128] A composition according to the invention can be applied to the face, body, décolleté, in any form or vehicle known to those skilled in the art, in particular in the form of a solution, dispersion, emulsion, paste or powder.

[0129] In cosmetics in particular, applications can be proposed, in particular in the ranges of facial and / or body skin care and makeup-care ranges, in particular eyelashes and eyebrows.

[0130] The composition may also be incorporated into a non-woven or woven material, made of natural or synthetic fibers, wool, or any material intended to come into contact with the skin and which may be used in clothing, in particular tights and socks, shorts, day or night underwear, handkerchiefs, or fabrics, in order to to exert its cosmetic effect through this skin / textile contact and allow continuous topical delivery (cosmeto-textiles).

[0131] The formulas may be included in ranges of personal care products and / or beauty products, in particular ranges of skin care, cleansing, makeup, makeup removal, anti-sun, artificial tanning, pre-shave, shaving, or after-shave, moisturizer, humectant, emollient, conditioner, exfoliant, astringent, depilatory, antiperspirant or antiperspirant, deodorant, deodorant, etc.

[0132] According to preferential and advantageous characteristics, the composition according to the invention may further comprise one or more other active ingredients.

[0133] The CTFA ("International cosmetic ingredient dictionary & handbook published by "the Cosmetic, Toiletry, and Fragrance Association, Inc.", Washington, DC) describes a wide variety, without limitation, of cosmetic and pharmaceutical ingredients commonly used in the skin care industry, which are suitable for use as additional ingredients in the compositions according to the present invention, as long as they are physically and chemically compatible with the other ingredients of the composition and especially with the actives of the present invention. Furthermore, the nature of these additional ingredients must not unacceptably alter the benefits of the actives of the invention. These additional ingredients may be synthetic or natural such as plant extracts or come from a bio-fermentation process.

[0134] Other skin care active ingredients which are particularly useful in combination with the composition according to the invention can be found in the commercial documentation of Sederma, Crodarom and Alban Muller and on the website www.croda.fr.

[0135] Examples of commercial active ingredients include: betaine, glycerol, Actimoist Bio 2™ (Active organics), AquaCacteen™ (Mibelle AG Cosmetics), Aquaphyline™ (Silab), AquaregulK™ (Solabia), Carciline™ (Greentech), Codiavelane™ (Biotech Marine), Dermaflux™ (Arch Chemicals, Inc), Hydra'Flow™ (Sochibo), Hydromoist L™ (Symrise), RenovHyal™ (Soliance), Seamoss™ (Biotech Marine), Argireline™ (trade name for Lipotec's acetyl hexapeptide-3), spilanthol or an extract of Acmella oleracea known as Gatuline Expression™ (Gattefossé), an extract of Boswellia serrata known as Boswellin™, Deepaline PVB™ (Seppic), Syn-AKE™ (Pentapharm), Ameliox™, Bioxilift™ (Silab), PhytoCellTec™Argan (Mibelle), Papilactyl D™ (Silab), Preventhelia™ (Lipotec), or one or more of the following active ingredients sold by Sederma: Subliskin™, Venuceane™, Moist 24™, Vegesome Moist 24™, Essenskin™, Juvinity™, Revidrat™, Resistem™, Chronodyn™, Kombuchka™,Chromocare™, Calmosensine™, Glycokin factor S™, Biobustyl™, Idealift™, Ceramide 2™, Ceramide A2™, Ceramide HO3™, Legance™, Intenslim™, Prodizia™, Beautifeye™, , Pacifeel™, Zingerslim™, Meiritage™, Sebuless™, Apiscalp™, Rubistem™, Citystem™, Neonyca™, NG Shea Butter Unsaponifiables™, Majestem™, Hydronesis™, Poretect™, Crystalide™, Amberstem™, Synchrolife™, Sylverfree™, Feminage™, Ameyezing™, Mel[o]stem™, or mixtures of these.

[0136] Among the extracts of plants (in the form of classical extracts or prepared by an in vitro method) that can be used as active additives, we can mention, in addition and in particular, the extracts of lierre, for example lierre grimpant (Hederia de Pleuderum), Buldera de Pleuder Bupleurum falcatum, arnica (Arnica montana L.), rosemary (Rosmarinus officinalis N.), willow (Calendula officinalis), sage (Salvia officinalis L.), ginseng (Panax ginseng), ginko biloba, millepertuis (Hyperycum fragment), aculecus L. elm (Filipendula ulmaria L.), orthosiphon (Orthosiphon stamincus Benth.), artichoke (Cynara scolymus), seaweed (Fucus vesiculosus), birch (Betula alba), green tea, cola nut (Cola nipida), chestnuts, Indian chestnuts, Basiau, Centaux heather, fucus, willow, piloselle, ash extracts, cangzhu extracts, cry-santhellum indicum extracts,of plants of the genus Armeniacea, Atractylodis platicodon, Sinnomenum, pharbitidis, Flemingia, of Coleus such as C. forskohlii, C. blumei, C. es-quirolii, C. scutellaroides, C. xanthantus and C. barbatus, as an extract of roots of Coleus barbatus, Ballo de Ballo, Gussio, Davallia, Terminalia, Barringtonia, Tréma, Antirobia, Cecropia, Argania, Dioscoreae as Dioscorea opposita or mexicain, extracts of Ammi visnaga, Siegesbeckia, in particular Siegesbeckia orientalis, plant extracts of the family Erica myculier, in particular Erica mytilles (Vaccinium angustifollium) of Arctostaphylos uva ursi, Aloe vera, plants containing sterols (especially phytosterols), Manjistha (extract of plants of the genus Rubia, in particular Rubia cordifolia), Guggal (extract of plants of the genus Commiphora, in particular Commiphora mukul), an extract of Commiphora mucus chamomile, purple clover, Piper methysticum (Kava Kava de Sederma),of Bacopa monieri (Bacocalmine™, Sederma) and fouet de mer, of Glycyrrhiza glabra, of murier, of melaleuca (tea tree), of Larrea divaricata, of Rhabdosia rubescens, of Euglena gracilis, of Fibraurea recisa hirudinea, of Chaparral sortia, of Enan flower, of Enan chlorantha, of Mitracarpe of the genus Spermacocea, of Buchu barosma, of Lawsonia inermis L., of Adiantium capillus-veneris L., of Chelidonium majus, of Luffa cylindrica, of "Japanese Mandarin" (Citrus reticulata blanco var. unshiu), of Camélia Imperia, of Ginensis, of Ginensis, flavum, of Cupressus sempervirens, of Polygonatum multiflorum, of Loveyly hemsleya, of Sambucus nigra, of Phaseolus lunatus, of Centaurium, of Macrocystis pyrifera, of Turnera diffusa, of Anemarrhena asphodeloides, of Portulaca pilosa, of Humuxus, of capulus Arabica, of capulus paraguariensis, of Globularia cordifolia, , Oxydendron arboreum, Albizzia julibrissin, Zingiber zerumbet smith, Astragalus membranaceus, Atracty Iodes macrocephalae, Plantago lanceolata, Leontopodium alpinum (or eldelweiss), Mirabilis jalapa, Apium graveolens, Marrubium vulgare, Buddleja davidii Franch or orchids.

[0137] The suspension and / or the composition according to the invention can be applied locally to the targeted areas.

[0138] For example, for a cosmetic facial treatment, the European Cosmetics Directive has set a standard application quantity of 2.72 mg / cm2 / day / person for a cream and 0.5 mg / cm2 / day / person for a body lotion.

[0139] According to other particularities, the cosmetic treatment method according to the invention can be associated with one or more other treatment methods targeting the skin, such as for example treatments by light therapy, by heat or by aromatherapy.

[0140] According to the invention, it is possible to propose devices with several compartments or kits intended for the implementation of the method described above, and which could comprise, by way of example, and without this being limiting, in a first compartment a suspension according to the invention, and in a second compartment a complementary active ingredient, the compositions contained in said first and second compartments being considered here as combination compositions for simultaneous, separate or spread out use over time, in particular in one of the treatments defined above.

[0141] Preferably, according to the invention, the additional active ingredient is chosen from vitamin B3 compounds, including niacinamide, tocopherol, hexamidine, α-lipoic acid, resveratrol or DHEA, hyaluronic acid, retinol, which are active ingredients very commonly used in topical cosmetic or dermo-pharmaceutical compositions.

[0142] According to a fifth object, the present invention proposes the use of particles according to the invention as defined according to the first object, or of a suspension according to the invention as defined above according to the third object, or of a composition according to the invention as defined according to the fourth object, for a non-therapeutic cosmetic treatment of the skin and / or its appendages, by oral or topical route.

[0143] The present invention thus covers a non-therapeutic cosmetic treatment method for beautifying or improving the appearance and general condition of the skin and / or appendages and treating imperfections, in a subject who needs an effective amount of the suspension according to the invention or a composition comprising it, in a physiologically acceptable excipient.

[0144] The “effective” quantity according to the invention, that is to say its dosage in the composition depends on the intended use of the composition. It depends on various factors, such as age, patient condition, severity of the disorder. An effective amount means a non-toxic amount sufficient to achieve the desired effect.

[0145] These purely cosmetic treatments may be the same treatments as those recommended for peptides already on the market, or new treatments that have been identified. These treatments include in particular any type of anti-aging treatment, to prevent or treat the effects of photoaging, in particular wrinkles and fine lines, unsightly spots, signs of skin fatigue such as dark circles and bags, slimming treatments, a scalp and / or hair treatment.

[0146] According to a sixth object, the present invention provides particles according to the invention according to the first object, a suspension according to the invention as defined above according to the third object, or a composition according to the invention as defined according to the fourth object, for a therapeutic treatment, preferably topical. Such a treatment may be, for example, a topical anti-inflammatory treatment.

[0147] According to a seventh object, the present invention proposes the use of particles according to the invention according to the first object, of a suspension according to the invention as defined above according to the third object, as an active ingredient, for the manufacture of a physiologically acceptable composition suitable for the treatment of the skin and / or its appendages, by oral or topical route. Detailed description

[0148] The present invention will be better understood in light of the detailed description of exemplary embodiments and studies described below. Brief description of the drawings

[0149] - [Fig.l] schematically illustrates the morphology of the particles according to the invention;

[0150] - [Fig.2] is a Differential Scanning Calorimetry (DSC) analysis graph encapsulated Pal-KTTKS according to the invention;

[0151] - [Fig.3] is a graph of the kinetics of digestion by trypsin of a suspension of encapsulated Pal-KTTKS according to the invention;

[0152] - [Fig.4] is an image obtained by electron microscopy in transmission (MET) of a suspension of encapsulated Pal-KTTKS according to the invention;

[0153] - [Fig.5] is an image obtained by atomic force microscopy (AFM) of a suspension of encapsulated Pal-KTTKS according to the invention;

[0154] - [Fig.6] is an image obtained by transmission electron microscopy (MET) of a suspension of encapsulated Pal-GHK according to the invention;

[0155] - [Fig.7] is a graph illustrating the release rate of encapsulated Pal-GHK at 100 ppm versus time relative to that of free Pal-GHK; and

[0156] - [Fig.8] is a graph illustrating the release rate of encapsulated Pal-GHK at 1000 ppm versus time compared to that of free Pal-GHK.

[0157] A- Preparation of capsule suspensions according to the process of the invention

[0158] Raw materials (all % by weight are expressed relative to the total weight of the mixture formed by the different phases): Wax: Cetyl Palmitate, e.g. CRODAMOL® CP (Croda).

[0159] Oil: mixture of C8 / C10 fatty acid esters, for example CRODAMOL® GTCC (Croda). Surfactant: TA1: Lauryl Glucoside, for example PLANTACARE® 1200 UP (BASF); Peptides: pure in the form of di-, tri-, tetra-, penta-, hexa-peptide powders, possibly acylated on the N-terminal side, in particular Palmitoylated (Pal) or with a biotinyl in derivatization on the N-terminal side.

[0160] Optionally: a second surfactant TA2, which here is hydrogenated lecithin, for example P75-3™ (LIPOID).

[0161] Optionally: a co-solvent for the peptide, such as glycerin or sorbitol.

[0162] Optionally: added in one of phases A to C (see table below) or via additional phases:

[0163] pH adjuster: for example citric acid in a quantity suitable for having a pH of 5 to 6;

[0164] Bacteriostatic agent: 1 to 6%, for example 1,2-octanediol and / or pro-panediol;

[0165] Antioxidants: 0.01 to 1%, for example tocopherol.

[0166] Table 1 below gives examples of formulation according to the invention and following the table is the procedure for forming a suspension of capsules according to the invention. The values ​​correspond to % by weight relative to the total weight of the formula.

[0167] [Tables 1] Pal-P P Pal-G HK Biot-G HK Pal-KM O2K Pal-GQ PR Pal-KT FK Pal-KT TKS Pal-VGVA PG Phase A: Wax 2.5 2.5 2.5 2.5 2.5 2.5 2.5 2.5 Oil 2.5 / / / 4.5 4.5 4.5 / TA1 2.5 / 2.5 2.5 / 2.5 2.5 / Phase B: Oil 7.5 7.5 7.5 7.5 3 3 3 7.5 TA2 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 Phase C: Water Qsp 100 Qsp 100 Qsp 100 Qsp 100 Qsp 100 Qsp 100 Qsp 100 Qsp 100 Peptide 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.024 TA1 / 2.5 2.5 / 2.5 / / 2.5 Co-solvent / 10 / / 10 / / / pH Adjuster 0.05 0.05 0.03 0.05 0.07 0.01 0.01 0.24

[0168] Operating mode:

[0169] Step 1: parallel preparation of phases (A), (B) and (C); heating if necessary to a temperature suitable for dissolving the solid raw materials to make the phases substantially liquid and homogeneous. TA1 (here Lauryl glycoside) is either added to phase (A) or to phase (C), depending on the solubility of the peptide. Phase (A) containing the wax may contain part of the oil from phase (B) in order to help solubilize the peptide if necessary.

[0170] The co-solvent when present is added to the aqueous phase.

[0171] Step 2: hot mixing of the three phases (A), (B) and (C), at a temperature at which the three phases remain substantially liquid and homogeneous. Step 3: hot formation of an emulsion. Step 4: Reduction of particle size by homogenization under high hot pressure (3 cycles at 500 bar). Step 5: cooling to room temperature so as to form the suspension of submicron particles according to the invention. B- Characterization of the suspensions obtained 1- Methods used 1.1- Macroscopic characterization:

[0172] For each suspension, the presence or absence of heterogeneous aspects, phase separation or precipitate, and the color are visually estimated through a colorless transparent bottle without any instrument. The viscosity is also visually estimated in comparison with water. 1.2- pH:

[0173] The pH was measured directly in the formulation (without dilution) using a calibrated potentiometer (pH 4.0 and 7.0).

[0174] 1.3- Particle size:

[0175] Laser diffraction technique (Mastersizer 3000™, Malvern). The suspension is directly added into the wet dispersion unit containing water until obscuration in the range of 2 to 8%. The refractive index used is 1.456 corresponding to the index of wax, i.e. cetyl palmitate. The results are expressed as D50 corresponding to the particle diameters at the 50th percentile of the particle size distribution curve, representative of the average particle size.

[0176] Polydispersity: The distribution curve also shows the homogeneity of the particle size by calculating the index (or range): (D90-D10) / D50, D90 and D10 corresponding respectively to the particle diameters at the 90th percentile and 10th percentile of the particle size distribution curve. A range less than 2 is considered a narrow distribution, describing a homogeneity of the particle size. 1.4- Content of encapsulated peptide active ingredient:

[0177] It is quantified using an HPLC chromatography method, with a column, mobile phase and detection appropriate to each peptide active. An external calibration curve is produced.

[0178] Each suspension is treated by liquid / liquid extraction of a mixture composed of isopropanol, a buffer at pH=2-2.5 and an organic solvent (acetonitrile or methanol) depending on the peptide to be measured.

[0179] For the peptides Pal-KTTKS, Pal-GHK, Pal-GQPR and Pal-VGVAPG, the quantification is for example carried out by an HPLC method at 35°C with a Waters Symmetry™C18 5 pm 4.6x150 mm HPLC column, a mobile phase composed of a eluent water / orthosphoric acid / triethylamine and acetonitrile 99.8 / 0.1 / 0.1 nInIn. Detection is carried out at 210 nm.

[0180] For the Pal-PP peptide, the analysis is for example carried out at 40°C with a Macherey Nagel™ EC 125 / 4 NUCLEODUR 100-5 C8 ec column, a mobile phase composed of a water / orthosphoric acid / triethylamine and acetonitrile 99.8 / 0.1 / 0.1 v / v / v eluent. Detection is carried out at 210 nm.

[0181] For the Biot-GHK peptide, the analysis is carried out at 40°C with a Macherey Nagel™ CC 125 / 4 Lichrospher 100 RP 18 ec 5 pm column, a mobile phase composed of a 25 mM potassium dihydrogen phosphate buffer, pH=2, an ion extractor hexanesulfonic acid at 0.01% w / v in acetonitrile. Detection is carried out at 210 nm.

[0182] The precision and repeatability of the methods for dosing peptides allows a recovery greater than 90%.

[0183] 1.5- Zeta potential and particle size by dynamic light scattering (DLS):

[0184] Zeta potential was measured using Brookhaven PALS™ equipment on samples of encapsulated peptide suspension or placebo diluted 5000 times in water. Samples were stored at RT prior to analysis.

[0185] The Zeta potential (or electrokinetic Zeta potential) represents the potential difference between the surface of the particle, covered with opposing and firmly fixed ions, and the neutral point. It constitutes a good indicator of the interactions between particles and therefore of the stability of the colloids. A negative Zeta potential indicates a good "repulsion" between the particles of the suspension preventing the agglomeration of the particles. 2- Results

[0186] The suspensions obtained according to the process of the invention are homogeneous, without visible phase separation, of milky white color and without any visible heterogeneity. Their viscosity is low. The results are given in Table 2 below.

[0187] [Tables2] Pal-PP Pal-GH K Biot-GHK Pal-KMO2 K Pal-GQP R Pal-K TFK Pal-KT TKS Pal-VGV APG pH 8.43 5.25 4.87 5.78 5.98 4.92 5.26 5.6 D50 (pm) / 0.239 0.316 0.261 0.229 0.343 0.254 0.304 Polydispersity Index / 1.31 1.55 1.27 1.33 1.15 1.29 0.27 Dosage 100 PPm 105 PPm 107 PPm 78 ppm 96 ppm 61 PPm 100 PPm 260 ppm

[0188] The zeta potential was measured negative on a suspension of encapsulated Pal-GHK.

[0189] The results presented above show that by using similar formulations within the scope of the present invention, it was possible to encapsulate different peptides, varying the composition, number and sequences of amino acids. In all cases, the formulations were obtained with the desired particle size, polydispersity and encapsulation rate, and were approved in the day 0 analysis. Samples were then produced to be studied during stability at room temperature (RT) and at 40°C.

[0190] C- Proof of encapsulation and half-life time: digestion by trypsin 1- Principle and protocol:

[0191] The study was carried out on a suspension of Pal-KTTKS encapsulated according to the invention using trypsin, which is an enzyme that hydrolyzes a peptide whose sequence contains the amino acid lysine. The peptide is placed in the presence of trypsin and the remaining quantity of peptide, i.e. not hydrolyzed by trypsin, is measured as a function of time. Only the peptide released over time from the capsule is likely to be hydrolyzed.

[0192] At T0, the measurements will confirm that the peptide is well encapsulated.

[0193] The kinetics will then make it possible to determine the half-life time, which constitutes an indicator of the accessibility of the peptide to the enzyme, therefore an indicator of the protection of the peptide by the capsule.

[0194] Two solutions are compared: one containing the encapsulated peptide (100 ppm) and the other the free peptide (i.e., non-encapsulated) at 100 ppm as well. An acetate buffer solution at pH=5 containing 0.003% trypsin is added. The samples are heated to 25°C. The enzymatic reaction is stopped to carry out the measurements by adding isopropyl alcohol. The quantification of the peptide is carried out by HPLC according to the method described in paragraph B 1-4 above. 2- Results:

[0195] [Fig.3] is a graph illustrating the Pal-KTTKS content (%) as a function of time (min) at 25°C. In this figure, we see that the Pal-KTTKS content of the encapsulated Pal-KTTKS is reduced by 50% in 1045 minutes while for the free Pal-KTTKS the content decreases by 50% in 60 minutes. We can conclude that the Pal-KTTKS is indeed protected in the capsules, since it does not follow the same curve as the free, unencapsulated Pal-KTTKS.

[0196] The hydrolysis is monitored under the chromatographic analysis conditions described in paragraph B 1-4 above. On the chromatogram, we initially see a chromatographic peak of Pal-KTTKS at 10.5 min, then as the enzymatic degradation progresses, we see this peak reduce and two new peaks appear corresponding to Pal-KTTK at 10.26 min and Pal-K at 13 min.

[0197] The half-life values ​​Tl / 2 (i.e. loss of 50% of the initial content) are given in the following table 3:

[0198] [Tables3] Tl / 2 (min) from the graph in Figure 3 Encapsulated Pal-KTTKS 1045 Free Pal-KTTKS 60

[0199] These results show a half-life of encapsulated Pal-KTTKS more than fifteen times longer than that of free Pal-KTTKS. A longer half-life will confer more stability to the peptide in the skin because its degradation upon contact with skin enzymes will be slower. A greater quantity of intact peptide can be transported to the site of activity.

[0200] D- Encapsulation efficiency demonstrated by tangential flow filtration analysis 1- Principle and protocol:

[0201] Tangential flow filtration, also called cross-flow filtration, separates suspended particles by passing the suspension along the surface of a membrane. The Sartorius Vivaflow™ 50 R filtration membrane, with a cut-off threshold of 100 kDa MWCO, allows, on the one hand, the passage of the free peptide through the membrane, and on the other hand, the retention of the capsules. The liquid phase permeates through the membrane due to a pressure difference across the membrane. After filtration, a retentate (phase that has not passed through, containing the particles) and a filtrate (aqueous phase that has passed through) are recovered.

[0202] After cross-filtration, the free (unencapsulated) peptide is found in the filtrate while the encapsulated peptide cannot cross the membrane and is found in the retentate.

[0203] To determine the encapsulation efficiency, two comparative tests are carried out: a tangential filtration of a peptide suspension according to the invention and a placebo aqueous solution of free peptide at the same concentration as the encapsulated peptide. The quantity of free peptide of the different fractions, namely filtrate and retentate (including losses in the system) is measured by HPLC, according to the HPLC method described in paragraph B 1-4 above. 2- Results:

[0204] The test was carried out on a suspension of Pal-KTTKS according to the invention and on a suspension of Pal-GHK according to the invention.

[0205] [Tables4] Analyzed fraction Relative content of Pal-KTTKS (%) Relative content of Pal-GHK (%) Peptide Free form filtrate 98 95 retentate 5 6 Peptide Encapsulated form filtrate <LOD 7 rétentat 74 74

[0206] The results, expressed as a percentage of the quantity used, show:

[0207] For the free Pal-KTTKS,

[0208] - 98% of Pal-KTTKS crosses the membrane and is measured in the filtrate

[0209] For the encapsulated Pal-KTTKS,

[0210] - No Pal-KTTKS is found in the filtrate at the limit of detection (LOD (~ 2 ppm)),

[0211] - By adding the part lost on the system to the dosage of the retentate we obtain a yield of more than 74% of Pal-KTTKS transported in encapsulated form, not having crossed the membrane.

[0212] Similarly, for Pal-GHK, the results show:

[0213] For free Pal-GHK,

[0214] - 95% of Pal GHK crosses the membrane and is measured in the filtrate.

[0215] For the encapsulated GHK Pal,

[0216] - Pal-GHK is found in the filtrate, approximately 7%,

[0217] - By adding the part lost on the system to the dosage of the retentate we obtain a yield of more than 74% of Pal-GHK carried in encapsulated form, not having crossed the membrane.

[0218] This experiment provides proof of the effectiveness of the encapsulation, 88% of the Pal-KTTKS in the suspension according to the invention is found inside the capsules. E- Morphology of the particles:

[0219] 1- Differential Scanning Calorimetry DSC

[0220] An analysis was carried out at 10°C / min from 0 to 180°C on a suspension of Pal-KTTKS encapsulated according to the invention. The thermogram is given in [Fig.2]. It shows a single large endothermic peak around 50.09°C combined attributed to the mixture of wax and surfactants. This analysis confirms that we are in the presence of waxy particles, solid at room temperature, inside which the peptide active is encapsulated.

[0221] 2- Analysis by transmission electron microscopy (TEM):

[0222] A 5% diluted suspension in water is applied to a support and dried for 1 min. The sample was then stained with 2% uranyl acetate for 1 min.

[0223] An image of a sample of encapsulated Pal-KTTKS suspension is shown in [Fig.4]. Regularly shaped, substantially spherical particles are seen. The TA1 coating (Lauryl glucoside in this example) is visible around the sphere. The particle size is also consistent with the laser diffraction analyses presented above.

[0224] An image of an encapsulated Pal-GHK suspension sample is shown in [Fig.6]. It is also seen that the particles have a regular shape and are substantially spherical. The TA1 coating is also visible around the sphere. The particle size is also consistent with the laser diffraction analyses presented above.

[0225] 3- Analysis by atomic force microscopy (AFM)

[0226] AFM allows a surface to be analyzed point by point using scanning with a probe made up of a fine tip. It allows the observation of objects on a very small scale.

[0227] [Fig.5] shows an AFM image of a sample of Pal-KTTKS suspension encapsulated according to the invention (at 100 ppm). The image shows that the particles have been broken apart by the probe, showing that they are solid but remain flexible and deformable. The structure of the shell and core type particles is visible. F- Stability studies

[0228] For these studies, the encapsulated peptide formulations were stored in HDPE polyethylene bottles at the study temperature (room temperature and 40°C). At each analysis time, a new 30 g bottle is opened for the first time. All samples are handled under regular laboratory lighting and normal air conditions. No inert gas is used to fill vials or handle samples. 1- Stability at room temperature

[0229] The following encapsulated peptides were monitored: Pal-KTTKS, Pal-GHK, Pal-GQPR, Pal-KTFK, Pal-KMO2K, Pal-VGVAPG and Pal-PP. 1.1- Results for encapsulated Pal-KTTKS:

[0230] [Tables5] Analysis Time 0 1 month 3 months 6 months 12 months PH 5.26 5.42 5.41 5.41 5.98 D50 (pm) 0.254 0.263 0.280 0.292 0.276 Dosage (ppm) 100 89 81 85 89 1.2- Results for the encapsulated Pal-GHK:

[0231] [Tables] Analysis Time 0 1 month 3 months PH 5.55 5.62 5.63 D50 (pm) 0.298 0.385 0.424 Dosage ppm 105 107 72 1.3- Results for the encapsulated Pal-GQPR

[0232] [Tables?] Analysis Time 0 1 month 3 months PH 5.98 5.92 6.21 D50 (pm) 0.229 0.262 0.287 Dosage (ppm) 112 106 104 1.4- Results for encapsulated Pal-KTFK

[0233] [Tables8] Analysis Time 0 1 month 3 months 6 months pH 4.92 4.95 5.09 4.96 D50 (pm) 0.343 0.283 0.366 0.370 Dosage (ppm) 61 77 58 75 1.5- Results for encapsulated Pal-KMO2K

[0234] [Tables 9] Analysis Time 0 2 months pH 5.65 5.59 D50 (pm) 0.263 0.275 Dosage (ppm) 75.7 78 1.6- Results for encapsulated Pal-VGVAPG

[0235] [Tables 10] Analysis Time 0 3 months pH 5.6 5.9 D50 (pm) 0.304 0.295 Dosage (ppm) 240 238 1.7- Results for encapsulated Pal-PP

[0236] [Tables 11] Analysis Time 0 1 month 2 months pH 8.43 8.22 / D50 (pm) / / / Dosage (ppm) 100 106 102

[0237] All of these results show good stability at room temperature of the suspension according to the invention, the pH, the average size of the capsules, their polydispersity as well as their content of encapsulated peptide change slightly but within acceptable limits. There is no indication of degradation or destabilization of the capsules.

[0238] For Pal-KTTKS, stability of up to 1 year at room temperature has been observed. The formula is stable, capable of maintaining the capsule size and the amount of encapsulated peptide. For comparison, Matrixyl® containing unencapsulated Pal-KTTKS should be stored at approximately 5°C. 2- Accelerated stability

[0239] The study is carried out at 40°C. At this temperature, degradation is accelerated. There is a correlation with degradation at room temperature, 90 days at 40°C usually being equivalent to 1 year at room temperature. This study therefore constitutes a good indicator of the stability of the suspensions of encapsulated peptides according to the invention.

[0240] The following encapsulated peptides were monitored: Pal-KTTKS, Pal-GHK, Pal-GQPR and Pal-KTFK. 2.1- Results for encapsulated Pal-KTTKS

[0241] [Tables 12] Analysis Time and conditions 0 30 days at 40°C 90 days at 40°C PH 5.26 5.43 5.42 D50 (pm) 0.254 0.268 0.279 Dosage (ppm) 100 100 90 2.2- Results for encapsulated Pal-GHK

[0242] [Tables 13] Analysis Time and conditions 0 30 days at 40°C 90 days at 40°C PH 5.25 5.21 5.24 D50 (pm) 0.239 0.237 0.245 Assay (ppm) 105 109 72 2.3- Results for encapsulated Pal-GQPR

[0243] [Tables 14] Analysis Time and conditions 0 30 days at 40°C 90 days at 40°C PH 5.98 6.14 6.00 D50 (pm) 0.229 0.25 0.283 Assay (ppm) 112 112 92 2.4- Results for encapsulated Pal-KTFK

[0244] [Tables 15] Analysis Time and conditions 0 90 days at 40°C pH 4.92 4.97 D50 (pm) 0.343 0.366 Assay (ppm) 61 76 2.5- Results for encapsulated Pal-PP

[0245] [Tables 16] Analysis Time and conditions 0 60 days at 40°C Dosage (ppm) 100 102

[0246] All of these results show that the encapsulated peptides, even of different peptide sequences, are stable at room temperature for at least 3 months, and for Pal-KTTKS up to 1 year, and at least 3 months in accelerated stability at 40°C. G- Release profile

[0247] The mechanism of release by a combination of diffusion and erosion has been studied.

[0248] Diffusion through the wax of the capsules occurs naturally between areas of high and low concentrations. When the encapsulated peptide active is applied to the skin, a concentration gradient is formed from which the diffusion process begins.

[0249] To demonstrate the diffusion process, a sample of Pal-GHK encapsulated according to the invention at 100 and 1000 ppm is placed in a dialysis bag closed with clamps. The dialysis bag has a pore size smaller than that of the capsules, the capsules are therefore retained in the bag. The bag is placed in a control medium (water acidified with 0.066% citric acid with 3% tween 80, and 30% EtOH have been added to this solution) and stirred at a constant temperature. This stirring simulates general wear and erosion causing the release of the encapsulated contents then passing through the pores of the dialysis bag into the control medium. Samples of the medium are taken over time and analyzed by HPLC to confirm the presence and level of released Pal-GHK. This test demonstrates the ability of the particles according to the invention to deliver the encapsulated peptide active ingredient into the skin over a prolonged period of time.

[0250] Erosion occurs upon application to the skin or its appendages when the outer layer of the capsules begins to degrade resulting in the release of the peptide active. Diffusion and erosion will occur simultaneously, which accelerates the release of the active. The diffusion rate eventually decreases as the concentration gradient decreases, and then the peptide active will only be released by erosion.

[0251] Figures 8 and 9 show the time release profiles of Pal-GHK at 100 and 1000 ppm respectively compared to the free peptide.

[0252] As seen in these figures, the Pal-GHK contained in the capsule is released at a slower rate than the free (non-encapsulated) peptide. This allows us to conclude that the release is better controlled over time when the peptide is encapsulated according to the invention compared to the free non-encapsulated peptide, thus demonstrating the benefit of the encapsulation according to the invention and the ability to continuously deliver a benefit linked to the biological activity of the peptide active ingredient.

[0253] [Fig.8] shows the release of encapsulated Pal GHK without and with the second surfactant, as an example, hydrogenated lecithin. The figure shows that the addition of the second surfactant does not change the profile and is preferred for the formula for particle stability. H- Comparative studies

[0254] 1- Surfactant TA1 of alkylglucoside type of at least one saturated fatty alcohol in C10-C16: Comparison of lauryl glucoside and decyl glucoside (all other things being equal)

[0255] [Tablesl7] Pal-KTTKS Pal-GHK Formation of a suspension at T0 of submicron particles (D5O<1 pm) Stability over time at RT Formation of a suspension at T0 of submicron particles Stability over time at RT Lauryl glucoside Yes pH = 5.26 D50(pm) = 0.254 Encapsulated peptide = 100 ppm Yes (12 months) pH = 5.98 D50(pm) = 0.276 Encapsulated peptide = 89 ppm Yes pH = 5.20 D50(pm) = 0.250 Encapsulated peptide = 77 ppm Yes (3 months) pH = 6.11 D50(pm) = 0.243 Encapsulated peptide = 100 ppm Decyl glucoside Yes pH = 4.87 D50(pm) = 0.325 Encapsulated peptide = 69 ppm Yes (3 months) pH = 4.94 D50(pm) = 0.363 Encapsulated peptide = 83 ppm Yes pH = 4.98 D50(pm) = 0.274 Encapsulated peptide = 91.5 ppm Yes (3 months) pH = 5.08 D50(pm) = 0.319 Encapsulated peptide = 78 ppm

[0256] The two surfactants also make it possible to obtain a suspension of particles that are acceptable in terms of particle size (D50<0.6 pm) and in terms of stability over time (at least 3 months).

[0257] 2- Surfactant TA1 of the C10-C16 saturated fatty acid polyglycerol ester type and ethoxylated Polysorbate type: comparative results between lauryl glucoside, a mixture of polyglyceryl-4 laurate / sebacate and polyglyceryl-6 caprylate / caprate” (Natragem S140 from Croda) and Tween 80 (Polysorbate 80) (all other things being equal).

[0258]

[0259]

[0260]

[0261] [Tables 18] Pal-KTTKS Formation of a homogeneous suspension at T0 of submicron particles (D50clpm) Lauryl glucoside Yes pH = 5.26 D50(pm) = 0.254 Encapsulated peptide = 100 ppm Natragem S140 (1.75% relative to the total weight of the formula) Yes pH = 4.12 D50(pm) = 0.317 Encapsulated peptide =110 ppm Tween 80 (1.75% relative to the total weight of the formula) pH = 5.03 D50(pm) = 0.259 Encapsulated peptide = 99 ppm Natragen S140 and Tween 80 are suitable surfactants according to the invention, making it possible to obtain a suspension of particles acceptable in terms of particle size (D50<0.6 pm) and polydispersity. 3- Addition of the surfactant TA2 (in the example hydrogenated lecithin) [Table 19] Tests with Pal-GHK TA1 HLB>1 0 TA2 Lipoid P75-3 Formation of a homogeneous, stable suspension, without phase shift, without visible particles, of submicron particles at T0 Stability over time at 3 months at RT Stability over time at 3 months at 40°C (equivalent to 12 months at RT) X Yes pH = 4.93 D50(pm) = 0.264 Encapsulated peptide = 74 ppm Yes: pH = 4.93 D50(pm) = 0.353 Encapsulated peptide = 100 ppm No: Phase shift pH = 5 D50(pm) = 5.0 Encapsulated peptide = no dosage possible XX Yes pH = 5.29 Yes pH = 6.06 Yes pH = 5.24 D50(pm) = 0.239 Encapsulated peptide = 74 ppm D50(pm) = 0.247 Encapsulated peptide = 101 ppm D50(pm) = 0.245 Encapsulated peptide = 71 ppm

[0262] The addition of the second surfactant having an HLB<6 (hydrogenated lecithin in the example) makes it possible to maintain the size of the submicron particles over time. At 3 months at RT, the particle size is still acceptable without TA2 in the formula, and at the same time at 40°C, equivalent to 12 months at RT, the size is well above 1 micron and the dispersion is heterogeneous (2 phases). The results show that for the formation and stability at room temperature of the particles, a single surfactant TA1 is sufficient. On the other hand, for stability at temperatures higher than room temperature, the second surfactant T2 is recommended. 4- Peptide active ingredient content [Table 20] Test with Pal-GHK Formation of a suspension at T0 of submicron particles Stability over time at RT, at 1 month 100 ppm Yes D50(pm) = 0.318 Yes D50 pm) = 0.382 500 ppm Yes D50(pm) = 0.324 Yes D50(pm) = 0.341 1000 ppm Yes D50(pm) = 0.448 Yes D50pm) = 0.452 5- Choice of wax [Table 21] Test with Pal-KTTKS Formation of a suspension at T0 of submicron particles Cetyl Palmitate Yes pH = 5.15 D50(pm) = 0.27 Triglyceride esters of C18-C36 fatty acids (Syncrowax®) Yes pH = 5.61 D50(pm) = 0.392

[0265] Both types of waxes are suitable according to the invention. Cetyl type waxes palmitate are preferred because they reduce polydispersity. 6- Quantity of wax [Table 22] Test with Pal-KTTKS

[0266] Formation of a suspension at T0 of submicron particles Stability over time at RT (6 months) 2.5% wax Yes pH 5.26 D50(pm) = 0.254 Yes pH 5.41 D50(pm) = 0.292 5% wax Yes pH 5.45 D50(pm) = 0.307 Yes pH 5.74 D50(pm) = 0.342 7- Adding a co-solvent [Table 23] Test with Pal-GHK

[0267] Formation of a suspension at T0 of submicron particles Stability over time 1 month Without glycerin Yes pH 5.25 D50(pm) = 0.239 Yes pH 6.11 D50(pm) = 0.243 10% glycerin Yes pH 4.88 D50(pm) = 0.232 Yes pH 4.90 D50(pm) = 0.276 8- Type of oil [Table 24] Test with Pal-GHK

[0268] Formation of a suspension at T0 of submicron particles Caprylic / capric triglycerides Yes pH = 4.73 D50(pm) = 0.290 dosage = 99 ppm Ethyl oleate Yes pH = 6.08 D50(pm) = 0.300 dosage 119 ppm Triisostearin (triglyceride of isostearic acid) Yes pH 6.01 D50(pm) = 0.373 dosage 103 ppm

[0269] I- Example of encapsulation of a peptide according to the invention and adsorption of a second peptide

[0270] To the suspension obtained from encapsulated Pal-KTTKS particles (obtained from the formula described in Table 25 above containing 100 ppm of Pal-KTTKS), therefore once cooled to room temperature, 100 ppm of a second peptide, Pal-KTFK, pre-dissolved in water is added and the mixture is stirred. A suspension of particles is then obtained on the outside of which Pal-KTFK is adsorbed (release test presented below). The dosage of the two peptides is given in the table below.

[0271] [Table 25] Encapsulated Pal-KTTKS and adsorbed Pal-KTFK Peptide Analysis 3 months at 40°C Dosage (ppm) Pal- KTTKS 100 Pal- KTFK 78

[0272] The same tangential filtration experiment as described in paragraph Dl was carried out to show that the second peptide, Pal-KTFK, is adsorbed onto the particles and is not found in free form in the suspension.

[0273] The results obtained are described in the following table:

[0274] [Tables26] Analyzed fraction Relative content of Pal-KTFK (%) Adsorbed Pal-KTFK Filtrate 1.2 Retentate 82.5

[0275] The results, expressed as a percentage of the quantity used, show that: - 1.2% of Pal-KTFK is found in the filtrate; - Nearly 82.0% of Pal-KTFK is therefore well adsorbed by the particles and will be able to be advantageously “conveyed” by the particles jointly with the Pal- KTTKS encapsulated.

[0276] J- Example of encapsulation of a mixture of peptides according to the invention (co-encapsulation)

[0277] Raw materials (all % by weight are expressed relative to the total weight of the mixture formed by the different phases): Wax: Cetyl Palmitate, CRODAMOL® CP (Croda),

[0278] Oil: mixture of C8 / C10 fatty acid esters, CRODAMOL® GTCC (Croda), Surfactant TA1: Lauryl Glucoside (PLANTACARE® 1200 UP, BASF),

[0279] Surfactant TA2: Hydrogenated lecithin (P75-3, LIPOID),

[0280] Mixture of peptides: Pal-GHK and Pal-GQPR in pure form (powders),

[0281] Co-solvent: glycerin,

[0282] pH adjuster: citric acid,

[0283] Table 27 below gives the details of the formulation.

[0284] [Tables27] Phase A: %m / m Wax 2.5 TA1 / Phase B: Oil 7.5 TA2 0.5 Phase C: Water Qsp 100 Peptide Pal-GHK 0.01 Peptide Pal-GQPR 0.01 TA1 2.5 Co-solvent 5 pH adjuster 0.05

[0285] Procedure:

[0286] Step 1: parallel preparation of phases (A), (B) and (C); heating at a temperature suitable for dissolving the solid raw materials to make the phases substantially liquid and homogeneous.

[0287] Step 2: hot mixing of the three phases (A), (B) and (C), at a temperature at which the three phases remain substantially liquid and homogeneous. Step 3: hot formation of an emulsion. Step 4: Reduction of particle size by homogenization under high pressure while hot (3 cycles at 500 bar). Step 5: Cooling to room temperature so as to form the suspension of submicron capsules according to the invention.

[0288] We obtain the following results:

[0289] [Table 28] Co-encapsulation of Pal-GHK and Pal-GQPR Formation of a homogeneous suspension at T0 of submicron particles Stability over time, 1 month at RT Yes pH 5.58 D50(pm) = 0.268 Pal-GHK dosed at 83 ppm Pal-GQPR dosed at 79 ppm Yes pH 5.65 D50(pm) = 0.321 Pal-GHK dosed at 78 ppm Pal-GQPR dosed at 79 ppm

[0290] These results show that the method according to the invention makes it possible to encapsulate a mixture of two peptides in a stable manner over time. K- Examples of finished product formulations

[0291] Various formulations can be prepared, comprising as active ingredient a suitable percentage by weight of a suspension according to the invention encapsulating a peptide active, as prepared according to the examples described above.

[0292] These formulations may contain additional active ingredients, the latter possibly supporting and / or complementing the activity of the active ingredient according to the invention. These ingredients may be of any category depending on their function(s), the place of application (body, face, neck, bust, hands, scalp, hair, etc.), the desired final effect and the targeted consumer, for example anti-aging, moisturizing, firming, anti-redness, anti-stretch marks, sunscreen, pigmenting, etc. Formulation example 1: Cream form

[0293] [Tables29] Raw materials Name INCI Ingredient (% w / P) Phase A h2o Water Qsp 100 Carbopol Ultrez 10 ™ Carbomer 0.30 Phase B Brij S2-SS-(RB)™ Steareth-2 0.40 Brij SIO-SO-(RB)™ Steareth-10 1.20 Crodafos CES-PA-(RB)™ Cetearyl Alcohol (and) Dicetyl Phosphate (and) Ceteth-10 Phosphate 4.00 Crodacol CS90-PA-(RB)™ Cetearyl Alcohol 1.50 Crodamol AB-LQ-(RB) ™ C12-15 Alkyl Benzoate 1.50 Crodamol OSU-LQ-(JP) ™ Diethylhexyl Succinate 7.00 Phase C Glycerin Glycerin 2.50 Octanediol Caprylyl Glycol 0.50 Phase D Phenoxyethanol Phenoxyethanol qs Phase E Potassium Sorbate Potassium Sorbate qs Phase F h2o Water 5.00 NaOH 30% Sodium Hydroxide 0.75 Phase G Suspension according to the invention - 3.00

[0294] Procedure: Swell phase A without stirring for 30 min and put it Heat to 75°C in a bain-marie. Heat phase B to 75°C in a bain-marie, stirring well. Melt phase C and mix. Mix phase D into phase C, previously cooled. Pour phase C+D into phase A, stirring with a rotor-stator mixer at v=500 rpm. Mix well. Add phase B to the previous phase, stirring with a rotor-stator mixer at v=1000 rpm. Add phase E immediately to the previous phase, stirring with a rotor-stator mixer at v=1000 rpm. Mix well. Add phase F and mix well. Below 40°C, add phase G and mix well.

[0295] Examples of ingredients that can be added to this formulation (marketed by SEDERMA): a moisturizing ingredient such as OptimHyal® and / or an ingredient protecting against the harmful effects of blue light such as Synchrolife®. Formulation example 2: Shampoo form

[0296] [Tables30] Raw materials INCI name Function % w / w Phase A Empicol® ESB3 Sodium Laureth Sulfate Surfactant 20 Crodateric™ CAB 30 Aqua (and) Cocamidopropyl Betaine Surfactant 15 Incromine™ Oxide C Cocamidopropylamine Oxide (and) Aqua Surfactant 3 Crovol™ A70 PEG-60 Almond Glycerides Solubilizer 3 Phase B Water Aqua - qsp 100 Preservative - Preservative qs Sodium Chloride Sodium Chloride pH adjuster 2.00 Phase C Suspension according to the invention - Active 3.00

[0297] Procedure: Prepare well-homogenized phases A and B. Pour phase A into phase B while stirring. Homogenize carefully. Add phase C. Mix well.

Claims

Claims

1. Submicron particle comprising a peptide active ingredient consisting of a peptide or mixture of peptides and / or an amino acid or mixture of amino acids, characterized in that it is substantially spherical consisting of an oily core containing said peptide active ingredient surrounded by a wax shell which is solid at room temperature, at least one non-ionic surfactant having an HLB substantially greater than or equal to 10 forming an outer layer on said wax shell.

2. A particle according to claim 1, comprising a second peptide active adsorbed on said outer layer.

3. Particle according to claim 1 or 2, characterized in that it is in aqueous suspension.

4. Method for encapsulating a peptide active ingredient consisting of a peptide or mixture of peptides and / or an amino acid or mixture of amino acids, said method comprising the following successive steps: - a step of providing a mixture comprising: - an aqueous phase in which the peptide active ingredient is optionally solubilized using a water-miscible co-solvent and / or a pH adjuster, - a waxy phase which is solid at room temperature and essentially consisting of one or more waxes; and - an oily phase which is liquid at room temperature and essentially consisting of one or more oils, the mixture further comprising a system of at least one non-ionic surfactant (TA1) having an HLB substantially greater than or equal to 10 and each phase being made homogeneous and liquid, if necessary, by heating and / or by mechanical and / or physical means; and - a step of hot forming an emulsion of said mixture;- a step of reducing the size of the particles of said emulsion substantially to a submicron size; and - a cooling step so as to form a suspension of submicron structured lipid particles in which said peptide active ingredient is encapsulated.;

5. Particle according to one of claims 1 to 3, or method according to claim 4, characterized in that the ratio of the % by weight of wax relative to the % by weight of oil is between 1:1 and 1:

10.

6. Particle according to one of claims 1 to 3, or 5, or method according to Claim 4 or 5, characterized in that said first surfactant (TA1) is chosen from at least one alkylglucoside or one alkylpolyglucoside of at least one saturated C10-C16 fatty alcohol, and / or at least one polyglycerol ester of saturated C10-C16 fatty acid, and / or at least one polysorbate, one poloxamer and / or one polyvinyl derivative.

7. Particle according to one of claims 1 to 3 or 5 or 6, or method according to one of claims 4 to 6, characterized in that said first surfactant is chosen from lauryl glucoside, decyl glucoside, coco glucoside and their polyglucoside derivatives, or polyglyceryl 4 to 6 of lauric, sebacic, caprylic or capric acid, polysorbate 80, or a mixture thereof.

8. Method according to one of claims 4 to 7, characterized in that the first surfactant is added to the aqueous phase, solubilized if necessary by heating.

9. Particle according to one of claims 1 to 3 or 4 to 7, or method according to any one of claims 5 to 8, characterized by a second surfactant (TA2) having an HLB substantially less than or equal to 6.

10. Particle or method according to claim 7, characterized in that the second surfactant (TA2) is a phospholipid chosen from phosphatidic acid, a phosphatidylcholine or lysophosphatidylcholine, a glycerophosphocholine, a phosphatidylserine or a lysophos-phatidylserine, a phosphatidylethanolamine, a phosphatidylinositol or a sphingomyelin.

11. Particle or method according to claim 10, characterized in that the second surfactant is a hydrogenated phospholipid.

12. Method according to one of claims 5 to 11, characterized in that the second surfactant is added to the waxy phase, solubilized if necessary by heating.

13. Particle according to claim 1 to 3 or 5 to 7 or 9 to 12, or method according to one of claims 4 to 12, characterized in that the wax is chosen from cetyl palmitate, glyceryl tribehenate, glyceryl stearate or tristearate or a wax of plant or animal origin.

14. Particle according to claim 1 to 3 or 5 to 6 or 9 to 13, or method according to one of claims 4 to 13, characterized in that the oil is chosen from fatty acids essentially C8 to C18, mono-, di- or triesters of fatty acid(s) essentially C8 to C18, comprising mono-, di- or triglycerides of fatty acids, and / or fatty alcohols comprising C8 to C18 carbon chains.

15. Method according to any one of claims 4 to 14, characterized in that the percentage by weight of wax is from 1% to 15% relative to the total weight of the mixture or suspension.

16. Method according to claim 4 to 15, characterized in that the percentage by weight of wax is from 2% to 8% relative to the total weight of the mixture or suspension.

17. A method according to any one of claims 4 to 16, characterized in that the percentage by weight of oil is from 2% to 20% relative to the total weight of the mixture or suspension.

18. A method according to any one of claims 4 to 17, characterized in that the percentage by weight of oil is from 5% to 12.5% ​​relative to the total weight of the mixture or suspension.

19. Method according to any one of claims 4 to 18, characterized in that the percentage by weight of peptide active ingredient is between 0.001% and 0.5%, relative to the total weight of the mixture or suspension.

20. Method according to any one of claims 4 to 19, characterized in that the mixture comprises or consists of: - 1 to 5000 ppm of peptide active ingredient - 2 to 8% by weight of a wax or a mixture of waxes; - 7.5 to 12.5% ​​by weight of a vegetable oil or a mixture of oils; - 2 to 4% by weight of lauryl glucoside or decyl glucoside; - 0.5 to 2% by weight of phosphatidylcholine or hydrogenated phosphatidylserine; - optionally a co-solvent, a pH adjuster and / or a preservative; all percentages by weight being expressed as a function of the total weight of the mixture or suspension, and the remainder qsp to 100% by weight being provided by the weight of water.

21. Method according to any one of claims 4 to 20 comprising an additional step at the end of the method where a second peptide compound is added to the suspension of structured particles.

22. A method according to any one of claims 4 to 21 comprising a final step of drying the aqueous suspension so as to form a powder of particles.

23. Particle according to one of claims 1 to 3, or 5 to 7, or 9 to 14, or method according to one of claims 4 to 22, characterized in that the peptide active ingredient corresponds to the following general formula (A): X-(Xaa)nZ in which: - Xaa is an amino acid, derivative or analogue; - n an integer > 1, when n > 2 the amino acids being chosen independently of each other; - at the N-terminal end X chosen from H, -CO-R1, -SO2-R1 or a biotinoyl group; - at the C-terminal end Z chosen from OH, OR1, NH2, NHR1 or NR1 R2; and - R1 and R2 being, independently of one another, chosen from an alkyl, aryl, aralkyl, alkylaryl, alkoxy, saccharide and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and / or sulfur-containing, said group having from 1 to 24 carbon atoms and being able to have in its skeleton one or more heteroatoms 0, S and / or N; - the case where X=H and Z=0H being excluded.

24. Aqueous suspension of submicron structured lipid particles obtainable by the method according to one of claims 4 to 23.

25. Suspension according to claim 24, characterized in that the polydispersity rate is substantially less than or equal to 2 with an average particle size ranging from 0.1 to 0.9 pm.

26. Suspension according to claim 24 or 25, characterized in that the particles are structured substantially spherical and have a waxy envelope.

27. ​​Composition, in particular cosmetic for the treatment of the skin and its appendages, comprising as active ingredient an effective quantity of particles according to one of claims 1 to 3, 5 to 7, 9 to 14, or 23, or of a suspension according to claim 24 to 26 and a physiologically acceptable vehicle.

28. Use of particles according to one of claims 1 to 3, 5 to 7, 9 to 14, or 23, or of a suspension according to claim 24 to 26 or of a composition according to claim 27, for a non-therapeutic cosmetic treatment of the skin and / or its appendages, by oral or topical route.

29. Particles according to one of claims 1 to 3, 5 to 7, 9 to 14, or 23, or suspension according to claim 21 or 24 to 26 or composition according to claim 27, for therapeutic treatment.

30. Use of particles according to one of claims 1 to 3, 5 to 7, 9 to 14, or 23, or of a suspension according to claim 24 to 26 as an active ingredient for the manufacture of a physiologically acceptable composition suitable for the treatment of the skin and / or its appendages, by oral or topical route.

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