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Title:
NEW COSMETIC OR DERMOPHARMACEUTICAL TOPICAL USE OF A MIXTURE OF A GHK TRIPEPTIDE AND GQPR TETRAPEPTIDE
Kind Code:
A2
Abstract:
According to the invention, a mixture of A-GHK-B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), or their derivatives or analogs, for the therapeutical treatment of the papillary dermis, with: A = H, CO-R1, SO2-R1; B = OH, OR1, NH2, NHR1, NR1R2; R1 and R2 being, independently of one another, selected from an alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and/or sulfured, said group which may have a heteroatom in its skeleton including O, S and/or N; is used to prevent damage to the papillary dermis and/or repair the dermis papillary, to prevent and/or treat skin aging, in particular photo-induced aging.


Inventors:
FOURNIAL, Arnaud (53 Boulevard Murat, Paris, F-75016, FR)
MONDON, Philippe (53b avenue Carnot, Cachan, F-94230, FR)
Application Number:
IB2012/052681
Publication Date:
12/06/2012
Filing Date:
05/29/2012
Assignee:
SEDERMA (29 rue du Chemin Vert, Le Perray en Yvelines, F-78610, FR)
FOURNIAL, Arnaud (53 Boulevard Murat, Paris, F-75016, FR)
MONDON, Philippe (53b avenue Carnot, Cachan, F-94230, FR)
International Classes:
A61K8/64; A61K38/06; A61K38/07; A61P17/00; A61Q19/08
View Patent Images:
Domestic Patent References:
WO2009055663A1N/A2009-04-30
WO2006075311A1N/A2006-07-20
WO2006020646A1N/A2006-02-23
WO2005102266A1N/A2005-11-03
WO2005048968A1N/A2005-06-02
WO2004101609A2N/A2004-11-25
WO2004024695A1N/A2004-03-25
WO2003068141A2N/A2003-08-21
WO2003028692A2N/A2003-04-10
WO2002066668A2N/A2002-08-29
WO1994007837A1N/A1994-04-14
Foreign References:
200401326672004-07-08
200702377352007-10-11
DE102006046076A12007-04-19
Other References:
FRANCESCO LACARRUBBA ET AL: "Mesotherapy for skin rejuvenation: assessment of the subepidermal low-echogenic band by ultrasound evaluation with cross-sectional B-mode scanning", DERMATOLOGIC THERAPY, vol. 21, November 2008 (2008-11), pages S1-S5, XP055020933, ISSN: 1396-0296, DOI: 10.1111/j.1529-8019.2008.00234.x
SKIN RES. TECHNOL. vol. 15, 2009, pages 306 - 3132009
Claims:
CLAIMS

1. Topical use of a mixture of A-GHK-B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), or their derivatives or analogs, to prevent damage to the papillary dermis and/or repair the dermis papillary, with: A = H, CO-R1; S02-Ri; B = OH, OR1 ; NH2, NHR1 ;

NRiR2; Ri and R2 being, independently of one another, selected from an alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and/or sulfured, said group which may have a heteroatom in its skeleton including O, S and/or N.

2. Use according to claim 1 to prevent and/or treat aging of papillary dermis, in particular photo-induced aging.

3. Use according to claim 1 or 2 to reduce the fragmentation of the fribrillary network of the papillary dermis, in particular the SLEB (subepidermal low echogenic band).

4. Use according to the preceding claims, characterized in that the peptide(s) is subtituted at the N terminal end with an A group being an acyle CO-Ri, sulfonyle S02-Ri and B at the C terminal end is OH, OMe, OEt ou NH2.

5. Use according to anyone of the preceding claims, characterized in that Ri and/or R2 is an alkyl chain, preferably comprising 1 to 24 carbon atoms, preferably linear.

6. Use according to the preceding claim, characterized in that Ri and/or R2 are preferably lipophilic chains comprising 6 to 24 carbon atoms.

7. Use according to anyone of the preceding claims, characterized in that A is an acyle group (A = CO-Ri) in particular chosen among biotinoyl, acetyl, palmitoyl, elaidoyl, myristoyl, octanoyl, stearoyl, oleoyl and lipoyl.

8. Use according to anyone of the preceding claims, characterized in that the tripeptide is Pal- GHK-OH or Pal-GHK-NH2 and the tetrapeptide is Pal-GQPR-OH (SEQ ID NO: 3) or Pal-

GQPR-NH2 (SEQ ID NO: 4).

9. Composition comprising a mixture of A-GHK-B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), or their derivatives or analogs, for the therapeutical treatment of the papillary dermis, with: A = H, CO-Ri , SO2-R1; B = OH, OR1 ; NH2, NHR1 ; NRiR2; Rx and R2 being, independently of one another, selected from an alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and/or sulfured, said group which may have a heteroatom in its skeleton including O, S and/or N.

Description:
NEW COSMETIC OR DERMOPHARMACEUTICAL TOPICAL USE OF

A MIXTURE OF A GHK TRIPEPTIDE AND GQPR TETRAPEPTIDE TECHNICAL FIELD

The present invention relates to a new topical use, cosmetic or therapeutical, of a peptidic mixture comprising a GHK type tripeptide (hereafter symbolised A-GHK-B) and a GQPR type tetrapeptide (hereafter symbolised A-GQPR-B, SEQ ID NO: 1), for the treatment of skin (including scalp skin) of human or animal mammals. The invention aims for the cosmetic, hygiene and personal care, and dermopharmacy industries.

BACKGROUND ART

The GQPR (Glycine-Glutamine-Proline-Arginine, SEQ ID NO: 2) and GHK (Glycine-Histidine- Lysine) sequences belong to Matrikines, known to participate in the reconstruction of the extracellular matrix of the connective tissue.

A composition comprising a mixture of Pal-GHK tripeptide and Pal-GQPR tetrapeptide (SEQ ID NO: 3) in a physiologically acceptable excipient is marketed as anti- wrinkle active by the Applicant Sederma under the trade name Matrixyl™3000 (WO 2005/048968).

Dermis is composed of a part called deep reticular dermis and an upper part called papillary dermis which is in contact with the epidermis via the dermal-epidermal junction (DEJ). Compared to the reticular dermis, the fibre network of the papillary dermis is different, more fragile. The proportions of collagen I and III are different and the network contains more glycoproteoglycans and glycosaminoglycans. This area also contains fibroblasts that have a very important role in the formation of the epidermis. The papillary dermis forms thus an important area of the skin, but fragile, especially very sensitive to UV radiations.

The upper part of the papillary dermis is slightly echogenic and can thus be visualized by ultrasound imaging (dermal fibers less dense returning less echos). This part is thus also called the SLEB "Sub- epidermal Low Echogenic Band" (or also called SENEB, "sub-epidermal non-echogenic band"). It has been shown that the thickness of the papillary dermis SLEB increases with time and exposure to solar radiations.

The present invention aims to provide a composition adapted to act specifically and efficiently on the papillary dermis, especially for the treatment of skin affected by a photo-induced and/or chronological aging.

SUMMARY OF THE INVENTION

To this end, it is proposed the topical use, cosmetic or dermopharmaceutical, of a mixture of A-GHK- B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), or their derivatives or analogs, to prevent damage to the papillary dermis and/or repair the dermis papillary, with:

B = OH, OR1; NH2, NHRi, NRiR2, Ri and R2 being, independently of one another, selected from an alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and/or sulfured, said group which may have a heteroatom in its skeleton including O, S and/or N.

A is at the peptide N terminal end (conventionally at left).

B is at the peptide C terminal end (conventionally at right).

In vivo tests given after in the description show that thanks to the invention, papillary dermis can be reconstructed (by reducing the SLEB papillary dermis thickness and improving its density). Thanks to the invention, the papillary dermis can be rejuvenated.

Thus according to the invention, a mixture of A-GHK-B and A-GQPR-B (SEQ ID NO: 1) peptides, or of their derivatives or analogs, can be used for preventing and/or treating aging of the papillary dermis, especially for the treatment of a skin overexposed to solar radiations (photo-induced aging).

The application areas are multiple, wherever necessary, damaged areas and/or areas where the dermis is particularly sensitive, for example the lips. The eye contour and the neckline may be areas as well covered by the present invention uses. Uses can also be performed as a preventive mesure.

The topical use of the invention may be either of cosmetic or therapeutical type (dermopharmacy). For the therapeutic field, the present invention proposes a composition comprising a mixture of A- GHK-B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), or their derivatives or analogs, for the therapeutic treatment of the papillary dermis.

According to other features, the invention encompasses a composition comprising a mixture of A- GHK-B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), or their derivatives or analogs, for the therapeutic treatment of a skin over-exposed to radiations, especially solar radiations. The mixture of the invention will have a healing effect thanks to a papillary dermis reconstruction.

The peptides according to the invention may be optically pure, or consist of L or D isomers or a mixture thereof. The L isomers which are those present in nature may be preferred because less expensive.

In addition, the invention peptides may be fragments of a larger peptide, containing more amino acids than the tripeptide and tetrapeptide active "core".

The present invention also encompasses derivatives (with modifications and/or addition of a chemical function but no change in the carbon skeleton) and analogs (with modifications and/or addition of a chemical function but in addition a change in the skeleton carbon), complex with other species such as a metal ion (eg copper, zinc, manganese, magnesium, and others).

As known, the presence of groups other than H for A and/or other than OH or NH2 for B on the peptides may have several purposes including: - Modifying the lipophilic character of the peptide to improve its solubility in an excipient and/or its bioavailability and its ability to penetrate skin; and/or

- Associating a molecule having its own activity with the peptide (for example a vitamin).

According to other features of the invention:

- The peptide(s) is or are substituted at the N terminal end with A being an CO-Ri acyle or

S02-Ri sulfonyle group, and B at the C terminal end being OH, OMe, OEt or NH2; and/or Ri and/or R2 is an alkyle chain, preferably comprising 1 to 24 carbon atoms, preferably a linear chain; Ri and/or R2 are preferably lipophilic chains comprising 6 to 24 carbon atoms; Moreover A is preferably an CO-Ri acyle group, in particular chosen among biotinoyl, acetyl, palmitoyl, elaidoyl, myristoyl, octanoyl, stearoyl, oleoyl and lipoyl;

A = CO-Ri or S02-Ri, Ri being a polycyclic group comprising from 2 to 6 cycles, in particular derivated from oleanolic, ursolic, boswelllic, betulinic or madecassic acid;

Ri comprises a sugar part.

Preferably, A=Pal (Palmitoyl), Biot (Biotinoyl), Ac (Acetyl), Ela (Elaidoyl) and/or B=OH, NH2, OMet or OEt.

Examples of compound according to the invention are in particular:

- Pal-GHK-OH, Pal-GHK-OMet, Pal-GHK-OEt, Pal-GHK-NH2, Biot-GHK-OH, Biot-GHK- OMet, Biot-GHK-OEt, Biot-GHK-NH2, Ac-GHK-OH, Ac-GHK-OMet, Ac-GHK-OEt, Ac-GHK-NH2, Ela-GHK-OH, Ela-GHK-OMet, Ela-GHK-OEt and Ela-GHK-NH2.

- Pal-GQPR-OH (SEQ ID NO: 3), Pal-GQPR-OMet (SEQ ID NO: 4), Pal-GQPR-OEt (SEQ

ID NO: 4), Pal-GQPR-NH2 (SEQ ID NO: 4), Biot-GQPR-OH (SEQ ID NO: 5), Biot- GQPR-OMet (SEQ ID NO: 6) , Biot-GQPR-OEt (SEQ ID NO: 6), Biot-GQPR-NH2 (SEQ ID NO: 6), Ac-GQPR-OH (SEQ ID NO: 7), Ac-GQPR-OMe (SEQ ID NO: 8), Ac-GQPR- OEt (SEQ ID NO: 8), Ac-GQPR-NH2 (SEQ ID NO: 8), Ela-GQPR-OH (SEQ ID NO: 9), Ela-GQPR-OMe (SEQ ID NO: 10), Ela-GQPR-OEt (SEQ ID NO: 10) and Ela-GQPR-NH2 and (SEQ ID NO: 10).

Preferably the tripeptide is Pal-GHK-OH or Pal-GHK-NH2 and the tetrapeptide is Pal-GQPR-OH (SEQ ID NO: 3) or Pal-GQPR-NH2 (SEQ ID NO: 4), more preferably the mixture of Pal-GHK-OH and Pal-GQPR-OH (SEQ ID NO: 3) (= the mixture of Palmitoyle Tetrapeptide-7 and Palmitoyl Oligopeptide).

A topical composition for implementing the invention comprises an effective content of A-GHK-B tripeptide and A-GQPR-B tetrapeptide (SEQ ID NO: 1), as recited above, in a physiologically acceptable medium. The "effective" amount depends on various factors such as age, the state of the patient, the severity of the disorder or disease and the administration mode. An effective amount means a nontoxic amount sufficient to achieve the desired effect. In a cosmetic composition according to the invention, the peptides, to be present in an effective amount, are generally in proportions of between 0.000001% and 15% relative to the total weight of the composition, more preferably between 0.0001 % and 5%, depending on the destination of the composition and the desired effect more or less pronounced. The peptides may be present in the compositions according to the invention in variable relative proportions, in equivalent amounts, or otherwise in different proportions.

DETAILED DESCRIPTION

All percentages and ratios used herein are by weight of total composition and all measurements are made at 25 ° C unless otherwise mentioned.

"Physiologically acceptable medium" means according to the present invention, without limitation, an aqueous or hydroalcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a microemulsion, an aqueous gel, an anhydrous gel, a serum, a vesicle dispersion, a powder.

"Physiologically acceptable" means that the compositions are suitable for topical or transdermal use in contact with mucous membranes, nails, scalp, hair and skin of mammals and particularly human, compositions that 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 commonly called the excipient of the composition.

For example, the peptide mixture according to the invention may be solubilized in a hydroalcoholic matrix, for example consisting of a water/glycerin mixture.

The present invention also provides a cosmetic treatment method for repairing the papillary dermis comprising applying to the skin (including lips) of a mixture of peptides A-GHK-B and A-GQPR- B (SEQ ID NO: 1), or their derivatives or analogues, as defined above.

The treatment of the invention may be particularly interesting on the lips where the epidermis is very thin and the dermis very thick.

The peptide may be combined with other active ingredients at effective concentrations that can act synergistically or in reinforcement to achieve the desired effects on the papillary dermis described for the invention, such as the following agents: radiation filters, including UVA and UVB, moisturizing, calming, myorelaxant, slimming, restructuring, firming, acting on microcirculation, acting on inflammation, on free radicals, vitamins, etc.

The composition of the invention can be applied on the face, lips, eye contour, body, neckline, scalp, in any form or vehicles known to the skilled artisan, including in the form of a solution, dispersion, emulsion, paste or powder, individually or in pre-mixture or be conveyed individually or in pre-mixture by vectors such as macrocapsules, microcapsules or nanocapsules, macrospheres, microspheres, or nanospheres, liposomes, oleosomes or chylomicrons, macroparticles, microparticles or nanoparticles, macrosponges, microsponges or nanosponges, microemulsions or nanoemulsions, or adsorbed on powdery organic polymers, talcs, bentonites, spores or exines or any other inorganic or organic medium.

In cosmetics in particular, applications can be offered for example in the ranges of facial and body skin care treatments, makeup-care products, including lip products.

In general, the peptides of the present invention can be used in any form, in a bound form, incorporated or adsorbed on macro-, micro- and nano-particles, or on macro-, micro- and nano- capsules for the treatment of textiles, natural or synthetic fibers, wools, and all materials intended to come into contact with skin and that can be used in clothing, day and night underwears, handkerchiefs, or textiles, in order to exert its cosmetic or therapeutic effect via this contact skin/textile and allow continuous topical delivery.

A composition according to the invention may further contain other ingredients that can enhance the appearance and feel touch of the skin.

Examples of such agents can be found in the CTFA ("International cosmetic ingredient dictionary & handbook (13eme Ed. 2010) published by the « Cosmetic, Toiletry, and Fragrance Association, Inc.", Washington, D.C.) which discloses a non-limited wide variety of cosmetic and pharmaceutical ingredients conventionally used in the skin care industry that can be used as additional ingredients/compounds in the compositions for the present invention. Examples of these ingredient classes include, but are not limited to: healing agents, skin anti-aging agents, anti- wrinkle agents, anti-atrophy agents, skin moisturizing agents, skin smoothing agents, antibacterial agents, anti-parasitic agents, antifungal agents, fungicidal agents, fungistatic agents, bactericidal agents, bacteriostatic agents, antimicrobial agents, anti-inflammatory agents, anti-pruriginous agents, anesthetic agents, antiviral agents, keratolytic agents, free radicals scavengers, anti- seborrhea agents, antidandruff agents, the agents modulating differentiation, proliferation or pigmentation of the skin, penetration accelerating agents, desquamating agents, melanin synthesis stimulating or inhibiting agents, whitening, depigmenting or lightening agents, pro-pigmenting agents, self-tanning agents, NO-synthase inhibiting agents, antioxidants, free radical scavengers and/or agents against atmospheric pollution, reactive carbonyl species scavengers, anti-glycation agents, tightening agents, agents stimulating the synthesis of dermal or epidermal macromolecules and/or capable of inhibiting or preventing their degradation, such as for example collagen synthesis stimulating agents, elastin synthesis stimulating agents, decorin synthesis stimulating agents, laminin synthesis stimulating agents, defensin synthesis stimulating agents, chaperone synthesis stimulating agents, aquaporin synthesis stimulation agents, hyaluronic acid synthesis stimulating agents, fibronectin synthesis stimulating agents, sirtuin synthesis-stimulating agents, agents stimulating the synthesis of lipids and components of the stratum corneum (ceramides, fatty acids, etc.), collagen degradation inhibiting agents elastin degradation inhibiting agents, agents stimulating fibroblast proliferation, agents stimulating keratinocyte proliferation, adipocyte proliferation stimulating agents, melanocyte proliferation stimulating agents, keratinocyte differentiation stimulating agents, adipocyte differentiation stimulating agents, acetylcholinesterase inhibiting agents, glycosaminoglycan synthesis stimulating agents, DNA repair agents, DNA protecting agents, anti-itching agents, agents for the treatment and/or care of sensitive skin, firming agents, anti-stretch mark agents, astringent agents, sebum production regulating agents, dermo- relaxing agents, healing adjuvant agents, re-epithelialization stimulating agents, re-epithelialization co-adjuvant agents, cytokine growth factors, calming agents, anti-inflammatory agents, agents acting on capillary circulation and/or microcirculation, angiogenesis stimulating agents, vascular permeability inhibiting agents, agents acting on cell metabolism, agents able to improve dermal- epidermal junction, agents inducing hair growth, hair growth inhibiting or retardant agents, muscle relaxants agents, antipollution and/or anti-free radical agents, lipolysis stimulating agents, slimming agents, anti-cellulite agents, agents acting on the microcirculation, agents acting on the energy metabolism of the cells, cleaning agents, hair conditioning agents, hair styling agents, hair growth promoters, sunscreen agents, total sunscreen agents, make-up agents, detergents, pharmaceutical products, emulsifiers, emollients, organic solvents, antiseptic agents, deodorant actives, physiologically acceptable carriers, surfactants, abrasives, absorbents, aesthetic components such as fragrances, pigments, dyes, colorants, natural colorants, essential oils, touch agents, cosmetic astringents, anti-acne agents, anti-coagulation agents, anti-foaming agents, antioxidants, binders, biological additives, enzymes, enzymatic inhibitors, enzyme-inducing agents, coenzymes, chelating agents, plant extracts, plant derivatives, essential oils, marine extracts, agents obtained from a bio-fermentation or a biotechnological process, mineral salts, cell extracts, sunscreens (organic or mineral photoprotective agents active against ultraviolet A and/or B rays), ceramides, peptides, buffers, volumizing agents, chelating agents, chemical additives, colorants, cosmetic biocides, denaturants, medical astringents, external analgesics, film formers, such as polymers, for exacerbing film-forming properties and substantivity of the composition, quaternary derivatives, substantivity increasing agents, opacifying agents, pH adjusters and regulators (e.g. triethanolamine), propellants, reducing agents, sequestrants, decoloring and/or lightening agents, skin-conditioning agents (e.g., humectants, including miscellaneous and occlusive), moisture retaining agents, alphahydroxyacids, betahydroxyacids, moisturizers, epidermal hydrolytic enzymes, healing and/or calming agents, skin treating agents, anti-wrinkle agents, agents that reduce or treat bags under the eyes, exfoliating agents, thickeners, softening agents, gelling polymers, vitamins and their derivatives, wetting agents, peeling agents, soothing agents, curative agents of the skin, lignans, preservatives (i.e. phenoxyethanol and parabens), anti UV, cytotoxic agents, anti-neoplastics, viscosity modifiers, non-volatile solvents, pearling agents, anti-perspirant agents, depilatories, vaccine, perfumed water, skin restructuring agent (i.e. Siegesbeckia orientalis extract), excipients, charges, minerals, anti-mycobacterial agents, anti-allergenic agents, HI or H2 antihistaminics, anti-irritants, agents stimulating the immune system, agents inhibiting the immune system, insect repellents, lubricants, pigments or dyes, hypopigmentation agents, preservatives, light stabilizers, and mixtures thereof, as long as they are physically and chemically compatible with the other ingredients of the composition and especially with the active ingredients of the present invention. Also the nature of these additional ingredients should not unacceptably alter the benefits of the active ingredients of the invention. These additional ingredients can be synthetic or natural such as plants extracts, or come from a bio-fermentation process. Additional examples can be found in the CTFA Cosmetic Ingredient Handbook.

Such additional active ingredient/compound can be selected from the group consisting of: sugar amines, glucosamine, D-glucosamine, N-acetyl glucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetyl mannosamine, galactosamine, N-acetyl galactosamine, B3 vitamin and its derivatives, niacinamide, sodium dehydroacetate, dehydroacetic acid and its salts, phytosterols, salicylic acid compounds, hexamidines, dialkanoyl hydroxyproline compounds, soy extracts and derivatives, equol, isoflavones, flavonoids, phytantriol, farnesol, geraniol, bisabolol, peptides and their derivatives, di-, tri-, terra-, penta-, and hexapeptides and their derivatives, KTTKS (SEQ ID NO: 11), Pal-KTTKS (SEQ ID NO: 12), carnosine, N-acyl amino acid compounds, retinoids, retinyl propionate, retinol, retinyl palmitate, retinyl acetate, retinal, retinoic acid, water-soluble vitamins, ascorbates, C vitamin, ascorbyl glucoside, ascorbyl palmitate, magnesium ascorbyl phosphate, sodium ascorbyl phosphate, vitamin B and their salts and derivatives, Bl vitamin, B2 vitamin, B6 vitamin, B 12 vitamin, provitamins and their salts and derivatives, K vitamin and derivatives, pantothenic acid and its derivatives, pantothenyl ethyl ether, panthenol and derivatives, dexpanthenol, biotin, amino acids and their salts and derivatives, water soluble amino acids, asparagine, alanine, indole, glutamic acid, water insoluble vitamins, A vitamin, E vitamin, F vitamin, D vitamin and mono-,di-, and tri-terpenoids compounds, beta-ionol, cedrol, and their derivatives, water insoluble amino acids, tyrosine, tryptamine, particulate materials, butylated hydroxytoluene, butylated hydroxyanisole, allantoin, tocopherol nicotinate, tocopherol, tocopherol esters, palmitoyl-Gly-His-Lys (Pal-GHK), phytosterol, hydroxy acids, glycolic acid, lactic acid, lactobionic acid, keto acids, pyruvic acid, phytic acid, lysophosphatidic acid, stilbenes, cinnamates, resveratrol, kinetin, zeatin, dimethylaminoethanol, natural peptides, soy peptides, salts of sugar acids, Mn gluconate, Zn gluconate, piroctone olamine, 3,4,4'- trichlorocarbanilide, triclocarban, Zn pyrithione, hydroquinone, kojic acid, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl glucoside, pyridoxine, aloe vera, terpene alcohols, allantoin, bisabolol, dipotassium glycyrrhizinate, glycerol acid, sorbitol acid, pentaerythritol acid, pyrrolidone acid and salts, dihydroxyacetone, erythrulose, glycer aldehyde, tartaraldehyde, clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, witch hazel distillate, eicosene and vinyl pyrrolidone copolymer, iodopropyl butylcarbamate, a polysaccharide, an essential fatty acid, a salicylate, glycyrrhetinic acid, carotenoids, ceramides and pseudo-ceramides, a lipid complex, oils in general of natural origin such shea butter, apricot oil, onagre oil, prune oil, palm oil, monoi oil, hydroquinone, HEPES, procysteine, O-octanoyl-6-D-maltose, the disodium salt of methylglycinediacetic acid, steroids such as diosgenin and derivatives of DHEA, DHEA or dehydroepiandrosterone and/or a precursor or chemical or biological derivative thereof, N-ethyloxycarbonyl-4-para-aminophenol, blueberries extracts, phytohormones, extracts of the yeast Saccharomyces cerevisiae, extracts of algae, extracts of soyabean, lupin, maize and/or peas, alverine and its salts, in particular alverine citrate, extract of butcher's broom and of horse chestnut, and mixtures thereof, a metalloproteinase inhibitor.

Further skin care and hair care active ingredients that are particularly useful combined with the composition can be found in SEDERMA commercial literature and on the website www. sederma.fr.

The following commercial actives can also be mentioned, as examples: betain, 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), Essenskin™ (Sederma), Moist 24™ (Sederma), Argireline™ (trade name of the acetyl hexapeptide-3 of Lipotec), spilanthol or an extract of Acmella oleracea known under the name Gatuline Expression™, an extract of Boswellia serrata known under the name Boswellin™, Deepaline PVB™ (Seppic), PhytoCellTec™Argan (Mibelle), Papilactyl D™ (Silab), Preventhelia™ (Lipotec), Syn-AKE™ (Pentapharm), Ameliox™, Bioxilift™ (Silab), Juvinity™ (Sederma), Revidrat™ (Sederma), Chronodyn™ (Sederma), Venuceane™ (Sederma), Ceramides (Sederma) or mixtures thereof.

Among plant extracts which can be combined with the peptidic mixture of the invention, there may more particularly be mentioned extracts of Ivy, in particular English Ivy (Hedera Helix), of Bupleurum chinensis, of Bupleurum Falcatum, of arnica {Arnica Montana L), of rosemary {Rosmarinus officinalis N), of marigold {Calendula officinalis), of sage {Salvia officinalis L), of ginseng {Panax ginseng), of ginko biloba, of St.-John's-Wort {Hyperycum Perforatum), of butcher's- broom {Ruscus aculeatus L), of European meadowsweet {Filipendula ulmaria L), of big- flowered Jarva tea {Orthosiphon Stamincus Benth), of algae {Fucus Vesiculosus), of birch {Betula alba), of green tea, of cola nuts {Cola Nipida), of horse-chestnut, of bamboo, of Centella asiatica, of heather, of fucus, of willow, of mouse-ear, of escine, of cangzhu, of chrysanthellum indicum, of the plants of the Armeniacea genus, Atractylodis Platicodon, Sinnomenum, Pharbitidis, Flemingia, of Coleus such as C. Forskohlii, C. blumei, C esquirolii, C scutellaroides, C xanthantus and C Barbatus, such as the extract of root of Coleus barbatus, extracts of Ballote, of Guioa, of Davallia, of Terminalia, of Barringtonia, of Trema, of antirobia, cecropia, argania, dioscoreae such as Dioscorea opposita or Mexican, extracts of Ammi visnaga, of Siegesbeckia, in particular Siegesbeckia orientalis, vegetable extracts of the family of Ericaceae, in particular bilberry extracts (Vaccinium angustifollium) or Arctostaphylos uva ursi, aloe vera, plant containing sterols (e.g., phytosterol), Manjistha (extracted from plants of the genus Rubia, particularly Rubia Cordifolia), and Guggal (extracted from plants of the genus Commiphora, particularly Commiphora Mukul), kola extract, chamomile, red clover extract, Piper methysticum extract (Kava Kava™ from SEDERMA), Bacopa monieri extract (Bacocalmine™ from SEDERMA) and sea whip extract, extracts of Glycyrrhiza glabra, of mulberry, of melaleuca (tea tree), of Larrea divaricata, of Rabdosia rubescens, of Euglena gracilis, of Fibraurea recisa Hirudinea, of Chaparral Sorghum, of sun flower extract, of Enantia chlorantha, of Mitracarpe of Spermacocea genus, 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 Camelia sinensis, of Imperata cylindrica, of Glaucium 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 Humulus lupulus, of Coffea Arabica, of Ilex Paraguariensis, or of Zingimber Zerumbet Smith.

The compositions of the present invention may include other peptides, including, without limitation, the di-, tri-, terra-, penta-and hexapeptides and their derivatives. According to a particular embodiment, the concentration of the additional peptide, in the composition, ranges from lxl0"7 and 20%, preferably from lxl0~6% and 10%, preferably between lxl0"5% and 5% by weight. According to the present invention, the term "peptide" refers to peptides containing 10 amino acids or less, their derivatives, isomers and complexes with other species such as a metallic ion (e.g. copper, zinc, manganese, magnesium, and others). The term "peptides" refers to both natural peptides and synthetic peptides. It also refers to compositions that contain peptides which are found in nature, and/or that are commercially available.

Suitable dipeptides for use herein include but are not limited to carnosine (beta- AH), YR, VW, NF, DF, KT, KC, CK, KP, KK or TT. Suitable tripeptides for use herein include, but are not limited to RKR, HGG, GKH, GGH, GHG, KFK, KPK, KMOK, KM02K or KAvaK. Suitable tetrapeptides for use herein include but are not limited to RSRK (SEQ ID NO: 13) or KTFK (SEQ ID NO: 14). Non limitative suitable examples of pentapeptide include the KTTKS (SEQ ID NO: 11) and of hexapeptides the GKTTKS (SEQ ID NO: 15) and VGVAPG (SEQ ID NO: 16).

Other suitable peptides for use herein include, but are not limited to: lipophilic derivatives of peptides, preferably palmitoyl derivatives, and metal complexes as aforementioned (e.g. copper complex of the tripeptide HGG). Preferred dipeptide derivatives include Pal-beta-AH, Ac-YR- hexadecylester (Calmosensine™, Idealift™ from Sederma), Pal-KT, Pal-RT (Sederma). Preferred tripeptide derivatives include the copper derivative of HGG (Lamin™ from Sigma), Lipospondin (Ela-KFK) and its analogs of conservative substitution, Ac-RKR-NH2 (Peptide CK+), Pal-KM02K (Sederma) and derivatives thereof. A suitable tetrapeptide derivative for use according to the present invention includes, but not limited to, Ela-KTFK (SEQ ID NO: 17), (from Sederma), suitable pentapeptide derivatives for use herein include, but are not limited to, Pal-KTTKS (MATRIXYL™, SEQ ID NO: 12), Pal-YGGFX (SEQ ID NO: 18) with X being Met(M) or Leu(L) or mixtures thereof. Suitable hexapeptide derivatives for use herein include, but are not limited to, Pal-VGVAPG (SEQ ID NO: 19), Pal-GKTTKS (SEQ ID NO: 20) and derivatives thereof.

The preferred compositions commercially available containing a tripeptide or a derivative include Maxilip™, Biobustyl™ and Matrixyl™synthe'6™ of Sederma. The compositions commercially available preferred sources of tetrapeptides include Rigin™, Eyeliss™, Matrixyl™ Reloaded and Matrixyl 3000™ proposed by Sederma.

The following marketed peptides can be mentioned as well as additional active ingredients: Vialox™, Syn-ake™ or Syn-Coll™ (Pentapharm), Hydroxyprolisilane CN™ (Exsymol), Argireline™, Leuphasyl™, Aldenine™, Trylgen™, Eyeseryl™, Serilesine™ or Decorinyl™ (Lipotec), Collaxyl™ or Quintescine™ (Vincience), BONT-L-Peptide™ (lnfinitec Activos), Cytokinol™LS (Laboratoires Serobiologiques/Cognis), Kollaren™, IP2000™ or Meliprene™ (lnstitut Europeen de Biologie Cellulaire), Neutrazen™ (Innovations), ECM-Protect™ (Atrium Innovations), Timp-Peptide™, ECM Moduline™ (lnfinitec Activos).

Cosmetic treatment method

The present invention also proposes a cosmetic treatment method to improve the general appearance of the skin, thanks to a reconstruction of the papillary dermis, comprising the application to the skin of an effective amount of a composition according to the invention as recited above.

The composition according to the invention may be applied locally onto targeted areas. One of the major advantages of the present invention resides in the ability whenever necessary or desirable to be able to apply local selective "gentle" treatments through this topical, non-invasive method of application. The application can be carried out very locally using a liner, a pencil, a roll-on applicator, a brush, a micro-cannula, applicators which are well known to the skilled person.

According to other specific features, the topical cosmetic treatment method according to the invention can be combined with one or several other treatment methods targeting the skin such as luminotherapy, heat or aromatherapy treatments.

According to the invention, devices with several compartments or kits may be proposed to apply the method described above which may include for example and non-restrictively, a first compartment containing a composition comprising one of the peptide of the invention, and in a second compartment a composition containing the other peptide and/or excipient, the compositions contained in the said first and second compartments in this case being considered to be a combination composition for simultaneous, separate or stepwise use in time, particularly in one of the treatment methods recited above.

The treatment method of the invention is more particularly adapted to the treatment of photodamaged skin.

1. GALENIC

Different examples of galenic formulas are given below containing the Pal-GHK-OH peptide and the Pal-GQPR-OH peptide (SEQ ID NO: 3) according to the invention as active ingredient. These formulas can also incorporate additional active ingredients, the latter coming if appropriate to support and/or in addition to the activity of the active ingredient of the invention. These ingredients can be of any class according to their(s) function(s), the application site (body, face, hands, etc.), the desired final effect and the target consumer.

Active ingredients used according to the invention:

MATRIXYL® 3000 comprising:

Actives: Pal-GHK-OH (Palmitoyl Oligopeptide) + Pal-GQPR-OH (Palmitoyl Tetrapeptide-7, SEQ ID NO: 3).

Excipient: glycerin, eau, butylen glycol, carbomer and polysorbate.

MATRIXYL® 3000 Lipo comprising:

Actives: Pal-GHK-OH (Palmitoyl Oligopeptide) + Pal-GQPR-OH (Palmitoyl Tetrapeptide-7, SEQ ID NO: 3).

Excipient: Alkylated benzoate, sorbitan laurate, tristearin and acetylated glycol stearate.

RIGIN™ comprising:

Actives: Pal-GQPR-OH (Palmitoyl Tetrapeptide-7, SEQ ID NO: 3).

Excipient: glycerin, eau, polyethoxylated alcohol.

BIOPEPTIDE CL™ comprising:

Actives: Pal-GHK-OH (Palmitoyl Oligopeptide)

Excipient: glyceryl polymethacrylate and propylene glycol.

Example 1: Body and face cream

Ingredient INCI Name Weight %

Phase A

Demineralised water Water (Aqua) qsp 100

Ultrez 10 Carbomer 0.25

Phase B

Glycerin Glycerin 3.50

Phase C

Brij S2 SS Steareth-2 0.40

Brij S10 SO Steareth-10

1.20

Crodafos CES Cetearyl Alcohol (and) Dicetyl Phosphate

4.00 (and) Ceteth-10 Phosphate

BRB CM 56 Cyclopentasiloxane (and) 2.00 Cyclohexasiloxane

Laurocapram Laurocapram 2.50 Crodamol OSU Diethylhexyl Succinate 7.00

Conservatives qs

Phase D

Potassium sorbate Potassium Sorbate 0.10

Phase E

NaOH 30% Sodium Hydroxide 0.40

Phase F

[Glycerin & Water & Butylene Glycol &

MATRIXYL® 3000 Carbomer & Polysorbate 20 & Palmitoyl 3.00

Oligopeptide & Palmitoyl Tetrapeptide-7]

Procedure: Weigh phase A and let swell without stirring for 30 min. Put phase A heated to 75 ° C in a water bath. Weigh phase B. Weigh phase C and heated to 75 ° C in a water bath. Mix well. Add phase B into phase A. Pour phase C into phase A+B under staro stirring. Mix well. Add phase D, extemporaneously. Add phase E; homogenize well. Then add phase F; mix well.

Example 2: Body milk

Procedure: Weigh and mix phase A. Weigh phase B. Weigh and mix phase C. Pour phase B into phase A under stirring. Add Phase C, mix well. Adjust the pH around 6.00 with phase D; mix well for one hour. Add phase E, homogenize well. Add phase E, homogenize well.

Example 3: Restorative cream

Ingredient INCI name Weight %

Phase A

Optasense RMA-IS Sodium Polyacrylate & Caprylic / Capric 2.00

Triglyceride & Mineral Oil & Tri (PPG-3 Myristyl Ether) Citrate & Sorbitan Laurate

& Trideceth-6

Crodamol CSO Cetearyl Ethylhexanoate 5.00 Crodamol GTCC Caprylic/Capric Triglyceride 5.00

Phase B

Demineralised water Water (aqua) Qsp 100

Phase C

Glycerin Glycerin

Phenoxyethanol Phenoxyethanol 5.00qs

Phase D

Demineralised water Water (aqua) 2.20 NaOH 30% Sodium hydroxyde 0.22

Phase E

Glycerin & Water & Butylene Glycol & 3.00

MATRIXYL® 3000 Carbomer & Polysorbate 20 & Palmitoyl

Oligopeptide & Palmitoyl Tetrapeptide-7

Phase F

Fragrance 0.10

Procedure: weigh and mix phase A. Weigh phase B. Weigh and mix phase C. Add phase B into phase A under stirring. Add phase C, mix well. Adjust the pH around 6.00 with phase D; mix well for 1 hour. Add Phase E, homogenize. Add phase E, homogenize.

Example 4: Serum form

Ingredient INCI name Weight %

Phase A

Demineralised water Water (Aqua) Qs 100

Ultrez 10 Carbomer 0.25

Phase B

Glycerin Glycerin 3.50

Phase C

Potassium sorbate Potassium sorbate 0.10

Phase.D

Brij S10 SO Steareth-10 1.50

Crodafos CS 20 Acid Cetearyl Alcohol (and) Ceteth-20 3.50

Phosphate (and) Dicetyl Phosphate

Span 60 Sorbitan Stearate 0.40

DC 245 Cyclopentasiloxane 4.00

Crodamol OSU Diethylhexyl Succinate 2.00

Phenoxyethanol Phenoxyethanol 1.00

Phase E

Triethanolamine 99% 0.50

Demineralised water Water (Aqua) 4.00

Phase F

Glycerin & Water & Butylene Glycol & 3.00

MATRIXYL® 3000 Carbomer & Polysorbate 20 & Palmitoyl

Oligopeptide & Palmitoyl Tetrapeptide-7

Phase G

Fragrance 0.10 Procedure: Sprinkle Ultrez 10 in water. Let swell for 30 minutes. Add phase B into phase A. Heat phase A+B at 75 ° C in a water bath. Weigh phase D, heat to 75 ° C in a water bath and mix well. Pour phase D into phase A+B under staro stirring. Mix well and add phase C. Then neutralize with phase E at about 50 ° C. Extemporaneously add phase F and G at about 35 ° C.

Example 5: Lip balm

Procedure: weigh phase A and melt around 75 °C. Mix well. Weigh phase B; add it to phase A.

Mix well. Weigh phase C; add it to phase A+B. Mix well. Pour the hot mixture in a pot or tube. Let cool before closing.

Example 6: Lip gloss

Ingredient INCI name Weight %

Phase A

Conservatives 0.80

Mineral oil qs 100

Syncrowax ERLC C 18-36 Acid Glycol Ester 4.00

Syncrowax HGLC CI 8-36 Acid Triglyceride 16.00

Crodacol C 90 Cetyl Alcohol 2.00

Novol Oleyl Alcohol 15.00

SHEA BUTTER Butyrospermum Parkii 1.50

Phase B

Brown covapate 0.20

Orange covapate 0.40

White covapate 1.30

Pink covapate 1.50

Crodamol PTIS Pentaerythrityl Tetraisostearate 10.00

Phase C

Fragrance 0.10

Phase D

MATRIXYL 3000 Lipo™ C12-15 Alkyl Benzoate& Sorbitan 3.00

Laurate& Tristearin& Acetylated Glycol

Stearate& Palmitoyl 01igopeptide&

Palmitoyl Tetrapeptide-7 Procedure: weigh phase A and melt around 90°C. Mix phase B and then add it to phase A. Add phase C extemporaneously. Melt Phase D and add it to phase A+B+C. Homogenize. Pour into molds. Cool to room temperature.

Example 7: Lipstick

Procedure: weigh phase A and melt around 90°C. Add phase B and mix. Add phase C and mix. Add phase D, stir and pour into molds. Cool to room temperature and store in cold place before unmolding.

Example 8: Gel form for eye contour

Ingredients INCI name Weight %

Phase A

H20 Water QsplOO

Cetyl hydroxyethylcellulose Cetyl hydroxyethylcellulose 0.30

Phase B

Ultrez 10 Carbomer 0.40

H20 Water 20.00

Phase C

Glycerin Glycerin 3.00

Panstat Ethyl & Methyl & Propyl parabens 0.30

Phase D

Marcol 82 Mineral oil 4.00

Crillet 1 Polysorbate 20 1.00

Crodamol AB C12-15 Alkyl Benzoate 2.00

Pemulen TR2 ClO-30 Alkyl Acrylate cross polymer 0.30

Phase E

Potassium sorbate Potassium Sorbate 0.10 Phase F

H20 Water 5.00

NaOH ION Sodium Hydroxide 0.50

Phase G

Water (Aqua) (and) Glycerin (and)

RIGIN™ Steareth-20 (and) Palmitoyl 2.00

Tetrapeptide-7

Glyceryl Polymethacrylate (and)

BIOPEPTIDE CL™ Propylene Glycol (and) Palmitoyl 2.00

Oligopeptide

Procedure: Disperse phase A under stirring. Sprinkle Ultrez 10 in water; let swell 30 minutes. Heat phase C until completely dissolved. Mix phase A with phase B. Add phase C in phase B+A. Add phase D, under stirring, in phase A+B+C. Add phase E. Neutralize with phase F. Add phase G and mix.

Examples of other ingredients which can be added to this gel type formulation in one of the phase according to their hydrophobe or hydrophilic physical property, at a certain % according to their concentration and the desired effect:

RENO V AGE™: global anti-ageing active ingredient marketed by SEDERMA (WO2006/020646). 3% by weight of this ingredient can be for example added to the formulation in phase D.

SUBLISKIN™: active ingredient marketed by SEDERMA (WO2009/055663) which hydrates and smoothes the skin while allowing it to resist to external aggression. 3% by weight of this ingredient may for example be added to the formulation in the phase G.

DERM AX YL™ : anti-aging active ingredient marketed by SEDERMA (WO2004/101609) which smoothes wrinkles and repairs the skin barrier. 3% by weight of the ingredient may for example be added to phase G.

Niacinamide (B3vitamine), retinol, resveratrol, DHEA : anti-aging actives, in particular anti- wrinkles. 0,5 % by weight of retinol, of resveratrol or DHEA may be added to phase G. 10% in weight of niacinamide at 10% in water can be for example added to phase G.

Hexamidine: antibacterial active that may be added to phase G of the formulation at 0.5% by weight.

CHRONODYN™: active ingredient actif marketed by SEDERMA (WO2006/075311) that tones and firms the skin, which erases signs of fatigue. 3% of this ingredient may for example be added to phase G.

VENUCE ANE™ : active ingredient marketed by SEDERMA (WO2002/066668) that prevents visible signs of photoaging (spots, wrinkles, dryness ...), protects cell structures from damage caused by UV and strengthens skin integrity. 3% of this ingredient may for example be added to phase G. JUVINITY™: active ingredient marketed SEDERMA that reduces signs of aging on the face and neckline, smoothes wrinkles, restructures and densifies the dermis. 2% of this ingredient may for example be added to phase G.

O.D.A.white™: active ingredient marketed by SEDERMA (WO1994/07837) which lightens the skin by reducing melanin synthesis. 1% of that ingredient may for example be added to phase G. Tocopherol (E vitamine) or q-lipoic acide (ALA): active with anti-oxidant and anti-radical properties. 0.5% by weight may for example be added to phase G.

LUMISPHERE™ : active ingredient marketed by SEDERMA (WO04/024695). It is the combination of diacetylboldine (DAB) encapsulated in microcapsules of polymethylmethacrylate and titanium dioxide modified with manganese (Ti02Mn). The Ti02Mn gives the skin a unifying, mattifying and brightening effect, and DAB provides a physiological lightening. 4% of this ingredient may for example be added to the phase G of the formulation.

REVIDRAT™: active marketed by SEDERMA, which in particular improves the epidermis cohesion and its hydration. 2% of this ingredient may for example be added to phase G of the formulation.

HALOXYL™: active marketed by SEDERMA (WO2005/102266), which improves the eye contour by reducing dark circles. 3% of this ingredient may for example by added to phase G of the formulation.

EYELISS™: is an active marketed by SEDERMA (WO2003/068141) which helps preventing and fighting against the appearance of bags under the eyes. 3% of this ingredient may be added to phase G of the formulation.

HYDRERGY™: active marketed by SEDERMA (WO2003/02828692) long lasting hydratant which stimulates ATP synthesis. 3% of this ingredient may be for example added to phase G of the formulation.

1. In vivo studies

The below studies were performed with the cream of above example 1.

Principle

Two complementary methods were used in order to study the superficial area of the dermis:

a SLEB (SubEpidermal Low Echogenic Band) analysis by high resolution 50Mhz echography. - an analysis of the structure of the fibre clusters in this area by Confocal Laser Microscopy Protocole

Specific inclusion criteria for studies: women were selected based on an age of between 51 and 72 years old (average age 59 years old) and on a prior echography examination of their skin to confirm the existence of a sufficiently large anechogenic area of dermis to be visualised and demonstrating a certain degree of ageing. The volunteers were required to follow a wash-out period using the test placebo for 15 days before measurements began. Subjects were required not to make any change to their hormone status during the 3 months before the test and during the test (no change in contraceptive, replacement or curative hormone therapy) and they were only allowed to use the cosmetics provided during the study.

Type and duration of study: this clinical study was conducted blind using non-invasive 5 measurements on 28 volunteers who were randomised and who applied the cream according to the invention to half of their face and one of their forearms. The placebo cream was applied controlaterally. Both creams were applied twice daily, by massaging, for 2 months.

The synopsis of the study can be summarized according to the below diagram.

TO T 1 month T 2 months

Π ^ υ Echography Echography Echography

Confocal laser microscopy Confocal laser microscopy Confocal laser microscopy

Statistical tests used the Student t test or a non-parametric Wilcoxon test, if needed, both on paired series.

Safety: The clinical study was conducted under medical supervision and demonstrated that product5 tolerability by the volunteers was excellent.

1) SLEB Analysis by high resolution echography

When ultrasounds hit a tissue in the human body, they are reflected and return a signal or "echo". The intensity of the echoes is translated by the echograph into grey scale levels to construct a reliable anatomical indicator of the area being investigated. The software in the instrument can also0 allocate colours depending on the intensity of the echoes.

The study was set out to assess changes in depth and density of the SLEB on a site which is not extensively exposed to sunlight (internal surface of the forearm) and on a more exposed site (external surface).

A Dermascan C™ (Cortex) echograph equipped with a 50 MHz frequency probe was used to5 obtain images approximately 6 mm wide and 3 mm thick with a resolution of 25 x 60 μιη. A sequence of 100 successive images was recorded over 4 cm. Five representative images were extracted and analysed by image analysis. The SLEB is traced accurately over a width of 5.5 mm allowing its depth (in μιη) and density (in Grey Scale Levels or GSL) to be calculated automatically.

0 Table 1: Changes in thickness and density of the SLEB after applying the cream of the invention or placebo (Internal surface of forearm; 28 volunteers, n= 5 replicates)

SLEB thickness (μηι) SLEB density (GSL)

TO T lmonth T 2months TO T lmonth T 2 months

19.67

171 175 173 18.95 18.93

Placebo +

± 20 ± 20 ± 20 ± 2.40 ± 2.31

1.91 % change vs. TO 2.7% 1.2% -3.7% -3.8% Significance nds nds nds nds

18.88

Cream according to 176 111 159 19.65 21.03

+

the invention ± 30 ± 20 ± 20 ± 2.34 ± 3.11

2.67

% change vs. TO -2.8% -9.8% 4.1% 11.4% Significance p<0.05 p<0.01 p<0.05 p<0.01 Maximum -» -17% -» -23% -» 32% -» 44% Responders 68% 93% 61% 68%

Delta (Invention -

-5.5% -11% 7.8% 15.2%

Placebo)

p<0.01 p<0.01 p<0.01 p<0.01

Significance

The results show that applying the cream of the invention produces from one month of application a significant fall in SLEB depth of -5.5% (p<0.01) associated with a +7.8% increase in density (p<0.01). Remarkably, these results are increased by a factor of 2 after applications for two months, reaching -11% depth and +15% density in this area (p<0.01 in both cases).

Measurements taken from the external forearm, i.e. on a light-exposed area, confirm these results (table 2).

Table 2: Change in thickness and density of the SLEB after applying the cream of the invention or placebo (External surface of forearm; 28 volunteers, n= 5 replicates)

The external surface of the forearm which is exposed to sunlight deteriorates more than the internal surface, which is better protected from the sun. The SLEB is thicker (195 μιη versus 171 μιη for the internal surface) and less dense (GSL of 16.2 compared to 19.7 for the internal surface).

Applications of the cream according to the invention for 2 months has a very similar effect to the effect seen on the protected surface, with a significant improvement in thickness (-14.4%) and density (+15.1%), both of which were significant p<0.01.

From published data on approximately 400 volunteers of different ethnic origins (QUERLEUX and coll. , Skin from various ethnic origins and aging: an in vivo cross-sectional multimodality imaging study. Skin Res. Technol. 2009; 15: 306-3132009), the equation expressing the change in SLEB depth as a function of age could be calculated. Using this equation and the data of the invention, the improvements in depth obtained with the invention can be converted into a theoretical age gain, an indicator which is easier to understand.

2) Analysis of dermis fibre clusters by confocal laser microscopy

For this study, an instrument called Vivascope™ 1500 has been used. It comprises three laser beams (785 nm, 658 nm and 445 nm) reflected differently depending on the refractory index of the structures they hit.

The Vivascope™ can firstly "illuminate" a specific point in the skin (focusing illumination) and, in parallel, accurately detect the returning light (focusing detection). This device is called "confocal" for "conjugate focal planes".

The main feature of the instrument is that it produces a high-definition, horizontal and vertical image in order to obtain a genuine real-time biopsy of the skin, entirely non-invasively. The instrument therefore allows horizontal in situ "sections" of the skin (500 μιη x 500 μιη) to be obtained with a horizontal resolution of 1.25 μιη and vertical resolution of less than 5 μιη.

Aged dermal fibres lose their filamentous appearance and organisation into networks taking on a blurred fluffy appearance, indicating fragmentation and degradation.

The study examined changes in structure of the superficial dermis of the face at a depth of -70 μιη after applying the cream according to the invention. This level allows the observation of the extent of degradation and fragmentation of fibres depending on the person's age and ultra violet exposures Analysis of the images obtained revealed the extent of degradation and fragmentation of the papillary dermis depending on the person's age and ultra violet exposure he was subjected.

The images obtained were analysed by Confoscan™ software (Orion concept) which through a specific analysis algorithm isolates and measures the size (circumference) of fibres. The more the fibers are whole, unsegmented, the more the perimeter of fibre clusters is important (fibre network less fragmented).

Table 3: Change in fragmentation of dermis fibre clusters after application of the cream of the invention (28 volunteers, n=3 measurements)

Mean fibre circumference at a depth of approx. 70 μηι (pixels)

TO T 1 month T 2 months

Placebo 192.9 ± 36.3 201.5 1 38.3 194.3 + 33.7

% change vs. TO 4.5% 0.7%

Significance nds nds

Cream of the invention 179.1 ± 25.3 199.0 1 39.4 204.1 + 45.4 % change vs. TO 11.1% 13.9%

Significance p<0.05 p<0.01

Maximum * -» 64% -» 54%

Responders 64% 71%

Delta

6.6% 13.2%

(Invention - Placebo)

p=0.27 p<0.05

Significance

After application of the cream according to the invention, the dermal fibre structure was greatly improved in the papillary area (SLEB) with a significant increase in their circumference of +11.1% at T 1 month then +13.9% at T 2 months. In parallel, the placebo caused little or no change in this parameter.

The good results measured in situ with the Vivascope™ clearly indicate that the cream according to the invention acted precisely at the level of the skin dermal fibres of the papillary area to restore an improved structure to the fibre network (reduction of fragmentation).

Therefore, reduction of the fragmentation of the fibre network of the papillary dermis, in particular the SLEB (or SENEB) can be obtained with the use of the peptidic mixture of the invention.