AND COSMETIC USES

TECHNICAL FIELD

The invention relates to a topical composition comprising bioactive sulfated oligosaccharides obtained by enzymatic cleavage of polysaccharides from green algae of the ulva genus. Cosmetic uses of this composition are also the subject matter of the present invention.

The present invention thus concerns in particular the cosmetics industry that manufactures and/or uses products for the treatment of the skin, scalp, mucous membranes and appendages (such as hair, eyelashes, eyebrows, nails, body hairs) of mammals, animals or humans, to improve their appearance and/or global condition.

The cosmetics industry is increasing demand for new products, particularly for new active ingredients that are derived from plant material to come within the scope of a sustainable development approach in comparison with a synthetic or chemical way of manufacturing.

Algae are thus an abundant source of raw material valued in many fields such as food, pharmaceuticals and cosmetics.

In cosmetics, algae are already widely used but mainly for thickening or gelling properties of their extracts which contain mainly polysaccharides.

Polysaccharides form also a particularly interesting molecule class especially because they contain sequences similar to some bioactive oligomers.

The Applicant has found particular interest in green algae and more particularly in ulva genus, and in the ulva lactuca species. It is known that polysaccharides from algae of ulva genus include in particular osidic units of iduronic acid (IdqpA) and rhamnose 3 sulfate (Rha/*3 sulfate) having an interesting potential for therapeutic or cosmetic applications (WO2009/016275). The Applicant has found particular interest in the two types of ulva polysaccharides that can be extracted from green algae walls of the ulva genus and containing such osidic units:

1) Ulvan polysaccharide comprising two main sequences, referred to as " aldobiuronic acid sequences ":

- A first sequence [^4)- -D-GlcpA-(l -^4)-a-L-Rha/>3 sulfate-(l -»] _n ; and

A second sequence [->4)-a-L-IdqpA-(l ->4)- a-L-Rha/*3 sulfate-(l ->] _m .

In these sequences, the « GlcpA » units correspond to glucuronic acid units.

Within this polysaccharide, glucuronan type sequences can also be found, but in minority. These sequences contain enchainments of glucuronic acid units:

[^4)- -D-GlcpA"(l -»4)-P-D-Glq>A-(l -»] _x .

2) Glucuronan polysaccharide comprising in majority glucuronan osidic sequences containing enchainments of glucuronic acid units

[^4)- -D-GlcpA-(1 ^4)- -D-GlcpA-(l ^ ]x, as disclosed above. An enzyme from the lyase class is capable of causing cleavage of the glycosidic bond between the osidic units according to a β elimination reaction (or transelimination). A ulvan-lyase enzyme is adapted to cleave more particularly the osidic bond between G cpA and Rha/*3 sulfate and between IdopA and Rha/*3, whereas a glucuronan-lyase enzyme is more particularly adapted to cleave the osidic bond between owing:

This cleavage processus shows saccharidic molecule fragments, oligosaccharides, having at the created non-reducing terminal end, a heterocycle having an ethylenic bond between carbon 4 and carbon 5, this residue corresponding to a 4-deoxy-(hex-4-ene) pyranosyluronic acid. The apparition of this ethylene bond and the dosage of the formed products can be monitored by UV spectroscopy at 235nm.

Cleaved oligosaccharides consist of oligo-ulvans and oligo-glucuronans. The proportion of each depends on the proportion of each of the starting glucuronan andulvan polysaccharides contacted with the enzymatic mixture.

Cleavage processes of ulva polysaccharides are described consisting of:

- Providing a microorganism capable of producing an enzymatic mixture of lyases;

- Contacting the polysaccharides extracted from ulva genus algae with said enzymatic lyase mixture to cause cleavage of the osidic bonds, and

- Recovering the cleaved oligosaccharides.

In the above mentioned WO2009/016275 document, a microorganism referenced CNCM 1-3776 is used to produce an enzyme mixture having a double enzymatic activity, ulvan lyase and glucuronan lyase, allowing thus to obtain a mixture of oligo-glucuronans and oligo-ulvans having a polymerisation degree (DP) inferior to 25 and with good yields for industrial production. According to the present application, a DP corresponds to a diholoside unit (containing two oses). Thus the sequence [->4)-P-D-GlcpA-(l ->4)-a-L-Rha/73 sulfate-(l ->] _n corresponds to a DP of 1, comprising one glucose/rhamnose diholoside unit.

However, to date, no application in the cosmetics field of selected oligosaccharide fractions obtained by enzymatic cleavage with a lyase mixture of polysaccharides extracted fromu/va genus green algae has been concretely carried out and described.

The aim of the present invention is to overcome this lack.

SUMMARY OF THE INVENTION

The present invention provides a topical composition comprising:

- A mixture of bioactive sulfated oligosaccharides having a polymerisation degree (DP) between 2 and 10, and - A physiologically acceptable medium;

said oligosaccharides been obtained according to an enzymatic cleavage process according to a β- elimination reaction of polysaccharides extracted from green algae of the ulva genus comprising:

Aldobiuronic sequences [->4)-P-D-GlcpA-(l ->4)-a-L-Rha/*3 sulfate-(l ->] _n and [->4)-a- L-IdqpA-(l ->4)- a-L-Rhap3 sulfate-(l ->] _m ; and

- Glucuronan sequences [-»4)-β-ϋ-αςρΑ-(1 - 4)-β-ϋ-αςρΑ-(1 - ] _x ,

Wherein in said sequences, the GlcpA units corresponding to glucuronic acid units, Rha/*3 sulfate to rhamnose 3 sulfate units and Id pA to iduronic acid units,

Using an ulvane-lyase enzyme adapted to cleave the osidic bond between GlcpA and Rha/*3 sulfate units and between Id pA and Rha/*3 sulfate units, and using a gucuronan lyase adapted to cleave the osidic bond between two GlcpA units.

The polymerisation degree (DP) is measured by a ionic chromatography method. The mean weight of the fraction is 6 kD measured by CPC/DEDL (centrifugal partition chromatography) with a sulfatation content of 13 to 17%.

In vitro tests demonstrated that this selected oligosaccharide fraction has a biological activity particularly interesting on skin cells, in particular anti-inflamatory, anti-radical, de-pigmenting effects on molecules of the dermal extracellular matrix (such as collagen 1 , collagen 4 and fibronectin), on epidermal extracellular matrix (hyaluronic acid). These tests are detailed below in the description. The present invention thus encompasses the use of the topical composition as recited above for a non- therapeutic cosmetic treatment of the skin and its appendages, in particular a depigmenting and/or anti-radical and/or anti- inflammatory treatment and/or a treatment to stimulate the molecules of the dermal extracellular matrix and/or to stimulate the molecules of the epidermal extracellular matrix and/or a moisturizing and/or anti-aging treatment. In particular, via the stimulation of the synthesis of molecules of the dermal extracellular matrix, the loss of skin density and firmness, wrinkles and fine lines, can be treated, and via the stimulation of the molecules of the epidermal matrix, loss of hydration, in particular transepidermal water loss, can be treated. Specific skin diseases can also be aimed, such as stretch marks, redness, reactive skin. Skin treatments can include soothing, healing, etc. Preferably, for the composition according to the invention, polysaccharides from ulva lactuca algae are used. Also preferably, the process is limited to the cleavage of ulvan polysaccharides priorly removed from glucuronan type polysaccharides and thus comprising mostly aldobiuronic sequences, the oligosaccharide mixture obtained being thus enriched in oligo-ulvans.

Furthermore, to obtain the active fraction of oligosaccharides according to the invention, the obtaining process is preferably used consisting of:

- Providing a microorganism capable of producing a ulvan lyase and a glucuronan lyase;

- Contacting the polysaccharides extracted from algae of the ulva genus with said enzyme lyases in order to induce the cleavage; and

- To recover the cleaved oligosaccharides. Referring to the WO2009/016275 document, according to the invention the microorganism preferentially comes from the above mentioned bacterial strain referenced CNCM 1-3776, filed and recorded at the "Collection Nationale de Cultures de Microorganismes" (CNCM) at the Institut Pasteur, 25 rue du Docteur Roux, F-75724 Paris cedex 15 (FR), belonging to the University of Picardie Jules Verne, Chemin du Thil, F-80025 Amiens (FR).

Other micro-organisms can be used of the Flavobacteriaceae family, or Ochrobactrum and Rhizobium genus (including Rhizobium meliloti NCIMB 40472).

According to other advantageous features, the oligosaccharides can be used in combination with one or more additional active ingredients, preferably to provide a range of wider cosmetic properties. The additional active ingredients are used either to reinforce or to supplement activity.

The additional active ingredients can be for example selected from lightening agents, anti-redness, anti-spots, calming, for treating reactive or sensitive skins, UV sunscreens, moisturizing, humectant, exfoliating, smoothing, toning, firming, volumizing, anti-aging, anti-wrinkles and fine lines, improving the mechanical and elastic properties, skin radiance, detoxifying actives, anti-hair-growth, acting on cutaneous barrier, anti-acne, action of sebum secretion, matifying, unifying, antiinflammatory, anti-oxidant, anti-radical, anti-glycation, for eye contour (dark circles and under eye bags), promoting blood circulation, peptides, vitamins etc. These active ingredients can be obtained from plant materials such as plant extracts or products of plant cell culture or fermentation.

More specifically, the oligosaccharides of the present invention can be combined advantageously with oligo-glucuronans, in particular the acetylated oligo-glucuronans disclosed in the patent application WO20120067327 and sold under the trade name Subliskin™ by Sederma. They are oligomer compounds of D-glucuronic acid or D-glucuronate with a β (1-4) sequence according to the following formula (

each ring of glucuronic acid of formula (I) comprising an acetylation degree comprised between 8,7±0,5 and 9,2±0,5% in weight of 0-CO-CH ₃ groups with regard to the weight of the glucuronic acid ring and a polymerisation degree comprised between 18 and 19 ±2. These oligomer compounds act at the level of dermal and epidermal elasticity and increase the dermo-epidermal cohesion in order to fight against skin ageing, wrinkles, fine lines, tactile and/or visible skin discontinuities, loss of firmness, of elasticity and tonus, and to fight against cutaneous tissue deformability. They also act on skin hydration.

Conventional active ingredients such as B3 vitamin compounds like niacinamide and tocopherol, retinoid compounds such as retinol, hexamidine, a-lipoic acid, resveratrol or DHEA, peptides, including N-acetyl-Tyr-Arg-O-hexadecyl, Pal-VGVAPG (SEQ ID NO: 1), Pal-KTTKS (SEQ ID NO: 2), Pal-GHK, Pal-KM0 ₂ K and Pal-GQPR (SEQ ID NO: 3) can be combined with the oligosaccharides of the present invention.

Other examples of additional ingredients are given below in the detailed description.

The present invention also aims a topical treatment method of the skin and its appendages, comprising the topical application to the skin and/or the appendages of an effective amount of a composition according to the invention comprising the oligosaccharides as recited above.

"Topical treatment" means according to the invention an application that is intended to act where it is applied: skin, mucous membranes, skin appendages.

Improvements in the appearance and general condition of the skin, mucous membranes, and appendages can be obtained by topical application on a regular basis such as daily.

The topical composition is preferably applied once daily for a period of at least a week, but it can be applied during periods of 2, 4, 8 or 12 weeks. The topical composition is preferably applied to the face and neck, but can be applied to any part of skin requiring an aesthetic improvement, where the composition remains on the skin area to be treated, and preferably is not removed or flushed from the skin.

It is also to be understood that, as used herein, the terms treating and treatment include and encompass the reduction, improvement, progress, relief, and/or elimination of dermatological effects, including aging. The compositions of the present invention and the methods are suitable for use to treat skin conditions of the skin in many areas of the skin, including without limitation, the face, forehead, lips, neck, neckline, arms, hands, body, legs, knees, feet, chest, back, buttocks, and others.

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. In the case of anti-wrinkle use for example it may be applied very locally using a syringe or micro-canula.

According to other specific features, the cosmetic treatment method according to the invention can be combined with one or more other treatment methods targeting the skin such as lumino-therapy, heat or aromatherapy treatments.

According to the invention, devices with several compartments or kits may be proposed to achieve the method described above which may include for example and non-restrictively, a first compartment containing a composition comprising the oligosaccharides of the invention, and in a second compartment a composition containing another active ingredient 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 present invention will be better understood in the light of the following detailed description. DETAILED DESCRIPTION

A. Preparation of the oligosaccharides of the invention

The protocol used is the one disclosed in WO2009/016275.

Polysaccharide forming the starting substrat

A mixture of polysaccharides comprised in the walls of ulva lactuca genus algae is heat extracted. After elimination of the insoluble elements, the pH of the extract is lowered to 2 with diluted acid in order to form a precipitate comprising glucuronic acid residues. The solution is thus recovered and brought to a neutral pH around 7. The solution is then purified, for example by ultrafiltration and precipitated with alcohol. The glucuronan polysaccharide fraction is thus eliminated and an ulvan polysaccharide is obtained comprising essentially aldobiuronic sequences, the glucuronan sequence left being in minority and the one integrated within the ulvan polysaccharide. The aim is here to subject only the ulvan polysaccharide to enzymatic cleavage in order to recover oligo-ulvans in majority (a mixture enriched in oligo-ulvan) but it goes without saying that the process can also according to a variant be achieved on an ulvan and glucuronan polysaccharide mixture.

The mean molecular weight of the ulvan polysaccharide is determined by sterical exclusion chromatography coupled to a light diffusion detector in line with a refractometer (SEC-MALLS technolocy, for « Size Exclusion Chromatography Coupled to Multi-angle Laser-Light-Scattering »); this mean weight is 6.10 ⁵ .

Preparation of the bacterial strain

From a bacterial strain, here the strain referenced CNCM 1-3776, an enzymatic substance is prepared having potentially a double enzymatic activity: ulvan- lyase and glucuronan- lyase.

The strain is grown on a mineral minimal nutritive medium comprising: NaCl: 22g; Na ₂ S0 _4: 3.7g; KC1: 0.6g; KBr: O. lg; MgCl ₂ ,6H ₂ 0: lOg; CaCl ₂ ,2H ₂ 0: 2.94g; NaHC0 ₃ : 0.16g; NaN0 ₃ : 25mg; NaH ₃ P0 ₄ ,12H ₂ 0: 5mg and H ₂ 0 QSP 1 liter; at a pH of 7.2.

The ulvan polysaccharide as prepared above (2g/L) is added to this nutrient medium as a carbone source substrate.

The strain, capable of degrading this ulvan polysaccharide, is isolated on a petri dish containing the same nutrient medium supplemented with agar. After incubation, colonies of strain, which metabolize the substrate by degrading it according by β-elimination and which are the most developed, are gathered.

Preparation of the enzyme activity

The CNCM 1-3776 strain is inoculated in a 2 liters capacity bioreactor, filled with 1.5 liters of a mineral medium: NaCl: 22g; Na ₂ S0 ₄ : 3.7g; KC1 0.6g; KBr: O.lg; MgCl ₂ ,7H ₂ 0: 5mg; NaN0 ₃ : 25 mg; NaH ₃ P0 ₄ ,12H ₂ 0, H ₂ 0 qs 1 liter; at pH of 7.2 and supplemented with yeast extrac l g/L; peptone: 5 g/L and ulvan polysaccharide of (2g/L).

Incubation is preferably carried out between 25 and 35°C under stirring for an average duration of 48 hours. During incubation, the concentration of enzyme mixture (ulvan-lyase and glucuronan-lyase) in the culture medium of the strain increases and therefore the enzymatic activity of the culture medium increases as well.

The culture medium is then cleared of the bacteria after incubation followed by addition of ammonium sulfate (60% w/v) to precipitate the proteins contained therein. This is followed by centrifugation of the precipitated culture medium and recovering of the protein base which is then reintroduced into about 10 ml of a buffer solution of Tris/HCl 20 mM at pH 7.5.

Chromatography on anion exchange resin can then retrieve only the fraction of an enzymatic substance having in majority an ulvan lyase activity and in minority a glucuronan lyase activity. An enzymatic activity of 30.10 ^"3 μmol/ml/min was quantified by measuring the absorbance at 235 nm. Enzymatic cleavage

The enzyme substance obtained above is applied to a substrate solution at 50g of ulvan polysaccharide for 1 liter of enzymatic solution.

The degradation experience is carried out at 20-30°C for a 3 hours incubation period (theoretically ranging from 1 to 72h).

The chromatographic profile by SEC-MALLS technology described above allows monitoring the oligo-ulvan fraction expected according to the invention (its average mass).

Analysis of the product resulting from the degradation by size exclusion chromatography on P2 Biogel (refractometry detection) highlights a major fraction analyzed by ESI-Q/TF type mass spectrometry (for electrospray-quadrupole-time of flight). The oligosaccharides have a polymerization degree DP comprised between 2 and 10, an average weight of 6kD and a sulfate ratio of 16.6%.

B. In vitro Tests

The various following tests were conducted on the oligosaccharide mixture obtained in section A above, highlighting the cosmetic activity according to the invention, as well as on comparative fractions:

1) Measurement of the anti- inflammatory effect on normal human fibroblasts (NHF)

FHN are grown until confluence. At this stage, they are put in contact with the products to be tested for 24h, then the cell mat are irradiated (UVB 70mJ/cm2) and contacted again with the products for 24 hours. The amounts of synthesized PGE2, IL-6 and IL-8 are measured in the culture supernatants by ELISA assay.

2) Measurement of the effect on the epidermal extracellular matrix (ECM):

Human keratinocytes (HaCat) are cultured for 24h in plates. The cells are contacted or not with the products to be tested for 3 days. The culture supernatants are removed and the hyaluronic acid amount is assayed. Retinoic acid is used as a positive control.

3) Measurement of the anti-radical effect (DCFH intracellular ROS):

FHN are seeded in well plates. After 24 hours of attachment, the cells are contacted for 24h with the products. Products are removed and the mats are rinsed. Cells are loaded for 30min with a "DCFH- DA" probe which becomes fluorescent in the presence of ROS (reactive oxygen species). After rinsing, the cells were placed again in contact with the products and the stress agent (800μιη H ₂ O ₂ ) for 2 hours. A fluorescence reading is used to estimate the amount of ROS present in the cells.

4) Measurement of the effect on dermal MEC (FHN)

FHN are grown in plate for 24 hours. The cells are contacted or not with the products to be tested for 3 days. The synthesis of collagen 1, collagen 4 and fibronectin is assessed by ELISA.

5) Measurement of the depigmenting effect:

Mouse melanoma cells (B16) are cultured in plates in DMEMc medium (DULBECCO'S Modified Eagle Medium) + 10% FCS (fetal calf serum). After 24 hours of attachment, the cells are contacted with the products to evaluate in the same medium for 48 h. At the end of the contact, the culture media were removed and the total melanin was assayed by spectrophotometry.

Comparative tables 1 and 2 below show the results of the different tests:

Table 1 :

Table 2:

These results clearly show that the fraction of the invention comprising the oligosaccharides mixture having a DP (2-10) has a cosmetic activity while the other fractions show no cosmetic activity or little activity.

C. Preparation of a composition according to the invention

The expression "physiologically acceptable medium" means according to the present invention, without limitation, an aqueous or hydro-alcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a micro-emulsion, an aqueous gel, an anhydrous gel, a serum, a dispersion of vesicles or a powder. "Physiologically acceptable" means that the compositions are suitable for topical or transdermal use, in contact with mucous membranes, appendages (nails, hair, body hair), scalp and skin of mammals, particularly human, without risk of toxicity, incompatibility, instability, allergic response, and others. The "physiologically acceptable medium" forms what is commonly called the excipient of the composition.

The choice of excipient of the composition is made according to the constraints of the oligosaccharides (stability, solubility, etc.), and if necessary according to the dosage form intended further for the composition. The oligosaccharides of the invention may be incorporated into the composition by means of an aqueous solution, or be dissolved by the usual physiologically acceptable solubilizers, for example and without limiting to this list: ethanol, propanol, isopropanol, propylene glycol, glycerin, butylene glycol, or polyethylene glycol or any combination thereof. It may also be interesting to solubilize the extract with emulsifiers.

Preferably according to the invention, an aqueous or hydro-alcoholic solution is used as the physiologically acceptable medium.

According to the invention, the effective quantity of oligosaccharides, that is to say their dosage, depends on various factors such as the age, the condition of the patient, etc. An effective amount means a not toxic amount enough to achieve the desired effect, which the skilled person is able to adapt.

The topical compositions for the present invention are generally prepared by conventional methods well known to one skilled in the art. Such methods may involve a mixture of ingredients in one or more steps to obtain a uniform state, with or without heating, cooling, etc.

The different galenic forms that may contain the oligosaccharides of the invention can be all forms i.e. creams, lotions, milks or creams ointments, gels, emulsions, dispersions, solutions, suspensions, cleansers, foundations, anhydrous preparations (sticks in particular lip balm, body and bath oils), shower and bath gels, shampoo and hair care lotions, milks or creams for skin care or hair, cleansing lotions or milks, sunscreen lotions, milks or creams, artificial tanning lotions, milks or creams, pre shave, shaving or aftershave creams, foams, gels or lotions, makeup, lipstick, mascara or nail polish, skin essences, serums, adhesive or absorbent materials, transdermal patches, or emollient powders, lotions, milks or creams, sprays, body and bath oils, foundation basis, ointment, emulsion, colloid, compact suspension or solid, pencil, sprayable formulation, brushable, blush, red, eyeliner, lipliner, lip gloss, face or body powder, styling gels or mousses, nail conditioning, lip balms, skin conditioners, moisturizers, lacquers, soaps, exfoliants, astringents, depilatories agents, permanent waving solutions, antidandruff formulations, antiperspirant or antiperspirant compositions, including sticks, "roll-on" deodorants, air fresheners, sprays for the nose and etc. These compositions may also be in the form of lipsticks intended either to color the lips or to prevent them from chapping, or makeup for eyes, eyes- shadows and foundations for the face. The compositions for the invention can include cosmetics, personal care products and pharmaceutical preparations. A composition in the form of foam or in the form of aerosol compositions also comprising a pressurized propellant can be considered. The oligosaccharides according to the present invention may be used in the form of solution, dispersion, emulsion, paste, or powder, individually or as a premix or vehicled individually or as a premix in vectors such as macro-, micro-, or nanocapsules, macro-, micro- or , nanospheres, liposomes, oleosomes or chylomicrons, macro-, micro-, or nanoparticles or macro-, micro or nanosponges, micro- or nanoemulsions, or adsorbed on organic polymer powders, talcs, bentonites, spores or exines, and other inorganic or organic supports.

The oligosaccharides according to the present invention may be used in any form whatsoever, in a form bound to or incorporated in or absorbed in or adsorbed on macro-, micro-, and nanoparticles, or macro-, micro-, and nano-capsules, for the treatment of textiles, natural or synthetic fibers, wools, and any materials that may be used for clothing or underwear for day or night, handkerchiefs or cloths, intended to come into contact with the skin, to exert their cosmetic effect via this skin/textile contact and to permit continuous topical delivery.

Additional ingredients

The CTFA (International cosmetic ingredient dictionary & handbook (13th Ed. 2010)), published by the Cosmetic, Toiletry, and Fragrance Association, Inc., Washington, D.C.) describes 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 as long as they physically and chemically compatibles with the other ingredients of the composition and especially with the oligosaccharides of the present invention. Also the nature of these additional ingredients should not unacceptably alter the benefits of the oligosaccharides of the invention. These additional ingredients can be synthetic or natural such as plants extracts, or come from bio- fermentation process. 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), 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), Syn-AKE™ (Pentapharm), Ameliox™, Bioxilift™ (Silab), Subliskin™ (Sederma), Venuceane™ (Sederma), Moist 24™ (Sederma), Essenskin™ (Sederma), Juvinity™ (Sederma), Revidrat™ (Sederma), Resistem™ (Sederma), Chronodyn™ (Sederma), Kombuchka™ (Sederma), Chromocare™ (Sederma), Calmosensine™ (Sederma), Glycokin factor S™ (Sederma), Odawhite™ (Sederma), Lumisphere™ (Sederma) and mixtures thereof. Among other plant extracts which can be combined with the composition 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 Zingiber zerumbet sm., 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 DavalUa, 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 Luff a 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 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 metal 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 and which are found in nature, and/or 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, GHK, GKH, GGH, GHG, KFK, KPK, KMOK, KM0 ₂ K or KAvaK. Suitable tetrapeptides for use herein include but are not limited to RSRK (SEQ ID NO: 4), GQPR (SEQ ID NO: 5) or KTFK (SEQ ID NO: 6). Suitable pentapeptides include, but are not limited to KTTKS (SEQ ID NO: 7). Suitable hexapeptides include but are not limited to GKTTKS (SEQ ID NO: 8) and VGVAPG (SEQ ID NO: 9).

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 include for example N-Palmitoyl-beta-Ala-His, N-Acetyl-Tyr- Arg-hexadecylester (Calmosensine™, Idealift™ from Sederma). Preferred tripeptide derivatives include for example N-Palmitoyl-Gly-Lys-His, and Pal-Gly-His-Ly, (Pal-GKH and Pal-GHK from Sederma), the copper derivative of HGG (Lamin™ from Sigma), Lipospondin (N-Elaidoyl-KFK) and its analogs of conservative substitution, N-Acetyl-RKR-NH ₂ (Peptide CK+), N-Biot-GHK (from Sederma), Pal-KM0 ₂ K (Matrixyl Synthe6™ from Sederma) and derivatives thereof. Suitable tetrapeptide derivatives for use according to the present invention include, but are not limited to, N- pal-GQPR (SEQ ID NO: 3) (from Sederma), suitable pentapeptide derivatives for use herein include, but are not limited to, Pal-KTTKS (SEQ ID NO: 2) (available as Matrixyl™ from Sederma), Pal- YGGF-X (SEQ ID NO: 10) with X Met or Leu or mixtures thereof. Suitable hexapeptide derivatives for use herein include, but are not limited to, Pal- VGVAPG (SEQ ID NO: 1) and derivatives thereof. The mixture of Pal-GHK and Pal-GQPR (SEQ ID NO: 3) (Matrixyl™ 3000, Sederma) can also be mentioned.

The preferred compositions commercially available containing a tripeptide or a derivative include Biopeptide-CL™, Maxilip™, Biobustyl™, Procapil™ and Matrixyl™synthe'6™ of Sederma. The compositions commercially available preferred sources of tetrapeptides include Rigin™, Eyeliss™, Matrixyl™ Reloaded and Matrixyl 3000™ which contain between 50 and 500 ppm of Pal-GQPR (SEQ ID NO: 3) and an excipient, 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™ or ECM Moduline™ (lnfmitec Activos).