WO2013088370A2

WO2013088370A2 - Use of the protein mmp-12 in the prevention and/or treatment of sensitive skin

Info

Publication number: WO2013088370A2
Application number: PCT/IB2012/057239
Authority: WO
Inventors: Isabelle Castiel; Audrey Gueniche
Original assignee: L'oreal
Priority date: 2011-12-14
Filing date: 2012-12-12
Publication date: 2013-06-20
Also published as: WO2013088370A3; FR2984128A1

Abstract

The present invention relates to the cosmetic use of an effective amount of at least one active agent consisting of a polypeptide having at least 80% amino acid identity with a metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 indicated below, or a C-terminal fragment of this polypeptide, for preventing and/or treating sensitive skin, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid. The present invention is directed mainly toward proposing a novel active agent for preventing and/or treating sensitive skin.

Description

USE OF THE PROTEIN MMP-12 IN THE PREVENTION AND/OR TREATMENT OF

SENSITIVE SKIN

The present invention is directed mainly toward proposing a novel active agent for preventing and/or treating sensitive skin.

The present invention relates to the use, especially the cosmetic use, of an effective amount of at least one novel active agent for preventing and/or treating sensitive skin, especially by reinforcing the skin barrier function.

In general, sensitive skin is defined by a particular reactivity of the skin. In contrast with skin termed as "allergic", the reactivity of sensitive skin is not a matter of an immunological process, i.e. it does not take place solely on skin that is already sensitized, in response to the presence of an allergen. Its mechanism is said to be aspecific.

This cutaneous reactivity is generally reflected by the manifestation of signs of discomfort in response to an individual coming into contact with a triggering factor, which may have diverse origins. The triggering factor may be the application of a cosmetic product to the surface of sensitive skin, the intake of food, exposure to abrupt changes in temperature, to atmospheric pollution and/or to ultraviolet or infrared rays. Related factors such as the skin type also exist. Thus, sensitive skin occurs more frequently among dry skin types than among normal skin types.

The appearance of these signs of discomfort, which appear within minutes of the individual coming into contact with the triggering factor, is one of the essential characteristics of sensitive skin. These signs are essentially dysaesthetic sensations. The term "dysaesthetic sensations" means the more or less painful sensations experienced on an area of skin, such as stinging, tingling, itching or pruritus, heating, discomfort, tautness, etc. These subjective signs most often arise in the absence of visible chemical signs such as redness and desquamation. It is nowadays known that these cutaneous irritation and intolerance reactions are especially associated with a release of neuropeptides by the nerve endings of the epidermis and the dermis.

As stated previously, sensitive skin is different from allergic skin. Its reactivity is not a matter of an immunological process, and is generally reflected solely by dysaesthetic sensations.

For obvious reasons, the absence of visible signs makes the diagnosis of sensitive skin difficult. This diagnosis is usually based on questioning of the patient. This symptomatology also has the advantage of allowing sensitive skin, whether or not associated with dry skin, to be differentiated from contact irritation or contact allergy, for which, on the other hand, there are visible inflammatory signs.

Consequently, the development of products designed for sensitive skin required the availability of tools for evaluating the sensory reaction of the skin. The first tools were inspired from their very conception by the essential characteristic of sensitive skin, namely the presence of signs of discomfort induced by a topical application.

Thus, the "stinging test" with lactic acid was the first test proposed. It is performed by recording the stinging sensations reported by a volunteer after application of a 10% lactic acid solution to the sides of the nose. Individuals reporting moderate or strong stinging sensations are termed "stingers" and are considered as having sensitive skin. On account of this cutaneous sensitivity to the topical application of a product, these individuals are then selected to test "sensitive skin products".

More recently, to specifically activate the peripheral nerve endings, involved in the discomfort and known as nociceptors, which have recently been identified as being involved in sensitive skin, new tests were proposed that use, precisely, other discomfort inducers, for instance capsaicin.

This second type of test, described in patent application EP 1 374 913, also constitutes another tool that is particularly useful for diagnosing sensitive skin.

For the purposes of the present invention, the term "sensitive skin" covers irritable skin and intolerant skin.

Intolerant skin is skin that reacts with sensations of heating, tautness, tingling and/or redness, to various factors such as the application of cosmetic or dermatological products or soap. In general, these signs are associated with an erythema and hyperseborrhoeic or acneic skin, or even rosaceiform skin, with or without dry patches.

However, an entirely satisfactory solution for preventing and/or treating this skin type termed "sensitive" is still not available at the present time, and this problem is more particularly exacerbated when this sensitive skin is combined with dry skin.

Dry skin is essentially manifested by a sensation of tautness and/or tension and is often associated with a reduction in the level of skin moisturization and impairment of the barrier function, measured by the imperceptible water loss. There is thus still a need for novel active agents that are capable of exerting beneficial cosmetic or therapeutic action on sensitive skin.

There is also still a need for active agents that can improve the quality of the skin barrier in the case of sensitive skin.

There is also a need for novel active agents for reducing skin sensitivity.

There is also a need for novel active agents for reducing skin dryness.

There is also a need for novel active agents for reducing skin reactivity.

There is also a need for novel compositions that are effective for preventing and/or treating sensitive skin.

The object of the present invention is to satisfy these needs.

Thus, according to a first subject, the invention relates to the cosmetic use of an effective amount of at least one active agent consisting of a polypeptide having at least 80% amino acid identity with a metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 indicated below, or a C-terminal fragment of this polypeptide, for preventing and/or treating sensitive skin, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid.

The active agent according to the invention contributes towards improving the quality of the skin barrier with, as a consequence, a reduction in the dryness, the sensitivity and, in general, the reactivity of the skin, more particularly sensitive skin.

According to another of its aspects, the present invention relates to a polypeptide with at least 80% amino acid identity with the metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, or a C- terminal fragment of the said polypeptide, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid, for its pharmaceutical or dermatological use for preventing and/or treating sensitive skin.

In certain embodiments, the prevention and/or treatment of the various skin conditions defined in the present description excludes the prevention and/or treatment of scalp conditions, and especially excludes the prevention and/or treatment of dandruff conditions of the scalp and the prevention and/or treatment of bacterial or fungal infections of the scalp, including scalp infections with Malassezia sp. This is likewise the case as regards the prevention and/or treatment of sensitive skin. According to another of its aspects, the present invention relates to a cosmetic and/or dermatological composition for preventing and/or treating sensitive skin and the associated dysaesthetic sensations, comprising, in a physiologically acceptable medium, at least one active agent in accordance with the invention, in combination with an effective amount of at least one additional probiotic agent, especially such as described below, this additional agent being different from the active agent according to the invention.

According to the present invention, such a composition is intended to be used topically.

According to another of its aspects, the present invention relates to a process, especially a cosmetic process, for treating and/or preventing sensitive skin and the associated dysaesthetic sensations, as defined previously, in an individual, comprising at least one step of administering to the said individual, especially topically, at least an effective amount of at least one active agent according to the invention, consisting of a polypeptide having at least 80% amino acid identity with the metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, or a C- terminal fragment of the said polypeptide, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid.

As stated below, the compositions containing it are formulated to be compatible with the adopted mode of administration, in the present case topical application.

The term "physiologically acceptable medium" means a medium that is compatible with all keratin materials such as the skin, the scalp, the nails, mucous membranes, the eyes and the hair, or any other area of bodily skin, and especially with the skin. A physiologically acceptable medium is preferentially a cosmetically or dermato logically acceptable medium, i.e. a medium that has no unpleasant odour, colour or appearance, and that is entirely compatible with the route of administration under consideration, namely the topical route.

According to the invention, the terms "preventing" and "prevention" mean reducing the risk of occurrence or slowing down the occurrence of a given phenomenon, for instance, according to one aspect of the present invention, deterioration of the skin's barrier function. For the purposes of the invention, the term "effective amount" means the minimum amount that is sufficient to observe the occurrence of a desired effect, namely, for example, the increase in the skin's barrier function. Such an amount may be determined by any method known to those skilled in the art, for example by means of preliminary experimental tests.

Active agent derived from the metalloprotease MMP-12

The present invention relates to the use of an effective amount of an active agent consisting of a polypeptide having at least 80% amino acid identity with the metalloprotease MMP-12 of SEQ ID NO: 1 , SEQ ID NO: 2 and SEQ ID NO: 3 indicated below, or a C-terminal fragment of this polypeptide, as an active agent, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid.

The matrix metalloprotease MMP-12, or MMP-12, is a metalloprotease belonging to the family of matrix metalloproteases, which are enzymes that especially have the role of degrading the extracellular matrix of connective tissues, and especially of the skin. This family of enzymes plays an important role in a large number of essential physiological processes, such as morphogenesis, angiogenesis and tissue repair.

MMP-12, also known under the name macrophage lipase, is especially known for its role in degrading elastin.

However, when they are expressed aberrantly or excessively, these enzymes may lead to the appearance of pathologies.

Thus, the role of MMP-12 in a certain number of pathologies such as arthritis, emphysema (Hautamaki et al. Macrophage elastase is required for cigarette smoke-induced emphysema in mice. Science 277, 2002-2004 (1997)) and vascular diseases (Curci et al. Expression and localization of macrophage elastase (matrix metalloproteinase-12) in abdominal aortic aneurysms. J. Clin. Invest. 102, 1900-1910 (1998)) has been demonstrated.

However, to the Applicant's knowledge, an activity of MMP-12 in relation with sensitive skin has never been described in the prior art. For the purposes of the present invention, the "percentage of identity" between two amino acid sequences is determined by comparing the two optimally aligned sequences, by means of a comparison window.

The part of the amino acid sequence in the comparison window may thus comprise additions or deletions (for example "gaps") relative to the reference sequence (which does not comprise these additions or these deletions) so as to obtain an optimal alignment between the two sequences.

The percentage of identity is calculated by determining the number of positions at which an identical amino acid is observed for the two compared sequences, followed by dividing the number of positions at which there is identity between the two amino acids by the total number of positions in the comparison window, and then multiplying the result by 100 in order to obtain the percentage of amino acid identity between the two sequences.

The optimal alignment of the sequences for the comparison may be achieved by computer using known algorithms.

In an entirely preferred manner, the percentage of sequence identity is determined using the CLUSTAL W software (version 1.82), the parameters being set as follows: (1) CPU MODE = ClustalW mp (2) ALIGNMENT = " full "; (3) OUTPUT FORMAT = " aln w/numbers " (4) OUTPUT ORDER = " aligned "; (5) COLOR ALIGNMENT = " no " (6) KTUP (word size) = " default "; (7) WINDOW LENGTH = " default "; (8) SCORE TYPE = " percent "; (9) TOPDIAG = " default "; (10) PAIRGAP = " default "; (11) PHYLOGENETIC TREE/TREE TYPE = " none "; (12) MATRIX = " default "; (13) GAP OPEN = " default "; (14) END GAPS = " default "; (15) GAP EXTENSION = " default "; (16) GAP DISTANCES = " default "; (17) TREE TYPE = " cladogram " and (18) TREE GRAP DISTANCES = " hide ".

A polypeptide or a C-terminal fragment thereof according to the invention may be synthesized via standard methods of synthetic chemistry, i.e. homogeneous chemical syntheses in solution or in solid phase. By way of illustration, a person skilled in the art may use the solution polypeptide synthetic techniques described by Houben-Weyl (1974, in Methode der Organischen Chemie, E. Wunsh ed., volume 15-1 and 15-11, Thieme, Stuttgart.). A polypeptide or a C-terminal fragment thereof according to the invention may also be synthesized chemically in liquid or solid phase by successive coupling of the various amino acid residues (from the N-terminal end to the C-terminal end in the liquid phase, or from the C-terminal end to the N-terminal end in the solid phase). A person skilled in the art may especially use the solid-phase peptide synthesis technique described by Merrifield (Merrifield R.B., (1965a), Nature, vol. 207 (996): 522-523; Merrifield R.B., (1965b), Science, vol. 150 (693): 178-185.)

According to another aspect, a polypeptide or a C-terminal fragment thereof according to the invention may be synthesized by genetic recombination, for example according to the production process comprising the following steps:

(a) preparing an expression vector into which has been inserted a nucleic acid coding for the polypeptide or a C-terminal fragment thereof of the invention, the said vector also comprising the regulatory sequences necessary for the expression of the said nucleic acid in a chosen host cell;

(b) transfecting a host cell with the recombinant vector obtained in step (a);

(c) culturing the host cell transfected in step b) in a suitable culture medium;

(d) recovering the culture supernatant of the transfected cells or the cell lysate of the said cells, for example by sonication or by osmotic shock; and

(e) separating or purifying, from the said culture medium, or from the cell lysate pellet, the recombinant polypeptide or a recombinant C-terminal fragment thereof of the invention.

To purify a polypeptide or a C-terminal fragment thereof according to the invention that has been produced by host cells transfected or infected with a recombinant vector coding for the said polypeptide or a C-terminal fragment thereof, a person skilled in the art may advantageously use purification techniques described by Molinier-Frenkel (2002, J. Viral. 76, 127-135), by Karayan et al. (1994, Virology 782-795) or by Novelli et al. (1991, Virology 185, 365-376).

Preferably, the MMP-12 under consideration according to the invention is human and of sequence SEQ ID NO: 1 as defined in the present patent application.

According to one embodiment of the invention, a fragment of the C-terminal part of a polypeptide in accordance with the invention may also be used, on condition, however, that this fragment comprises the amino acid sequence KDXK.

K represents a lysine (abbreviated as Leu) and D represents an aspartic acid (abbreviated as Asp). The amino acid defined as being X in the amino acid sequence KDXK mentioned previously is chosen from glutamic acid (abbreviated as Glu or E) and aspartic acid (abbreviated as Asp or D).

Such a C-terminal fragment that is suitable for use in the invention may comprise a sequence with a length of 80 to 20 contiguous amino acids, or even 50 to 20 contiguous amino acids, and preferably 30 to 20 contiguous amino acids at the C-terminal end of said polypeptide, and which comprises the sequence KDXK.

According to one preferred embodiment, a C-terminal fragment of a polypeptide in accordance with the invention consists at least, contiguously and in this order, of at least the 8 amino acids preceding the sequence KDXK according to the invention, the sequence KDXK itself, and then at least the 8 amino acids following the sequence KDXK in a polypeptide in accordance with the invention.

According to one preferred embodiment of the invention, the C-terminal fragment of a polypeptide in accordance with the invention, i.e. having at least 80% amino acid identity with the metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 is chosen from the following sequences:

SEQ ID NO: 4: ARNQ VFLFKDDKY WLI SNLR

SEQ ID NO: 5: GRNQLFLFKDEKYWLINNLV

SEQ ID NO: 6: SRNQLFLFKDEKYWLINNLV

According to one preferred embodiment, at least the amino acids constituting the sequence KDXK are in beta form.

According to one embodiment of the invention, an active agent in accordance with the invention consisting of a polypeptide having at least 80% amino acid identity with the metalloprotease MMP-12 of sequence SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3 indicated below, or a C-terminal fragment of this polypeptide, is used in a composition, especially a cosmetic composition, as defined below.

An active agent according to the invention may be present in a composition in a content of between 0.0001%> and 30%> by weight, preferably between 0.001%> and 15% by weight and preferentially between 0.1% and 10% by weight relative to the total weight of the composition. According to one embodiment variant, an active agent in accordance with the invention may be included in a composition of the invention in the form of a lyophilized powder, where appropriate in a concentrated form. Composition

The present invention relates to a cosmetic and/or dermatological composition that is useful for preventing and/or treating sensitive skin and the associated dysaesthetic sensations, comprising, in a physiologically acceptable medium, at least one active agent as defined previously.

The dysaesthetic sensations in accordance with the invention may be chosen especially from stinging, tingling, itching or pruritus, burning, heating, discomfort and tautness of the skin.

Such a composition also comprises at least an effective amount of at least one additional probiotic agent especially as described below, this additional agent being different from the active agent according to the invention.

This additional agent may be chosen especially from ascomycetes such as Saccharomyces, Yarrowia, Kluyveromyces, Torulaspora, Schizosaccharomyces pombe, Debaromyces, Candida, Pichia, Aspergillus and Penicillium, bacteria of the genera Bifidobacterium, Bacteroides, Fusobacterium, Melissococcus, Propionibacterium, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus and Lactobacillus, and mixtures thereof.

As ascomycetes that are most particularly suitable for use, mention may be made in particular of Yarrowia lipolytica and Kluyveromyces lactis, and also Saccharomyces cerevisiae, Torulaspora, Schizosaccharamyces pombe, Candida and Pichia.

Specific examples of probiotic microorganisms are Bifidobacterium bifidum, Bifidobacterium infantis, Lactobacillus acidophilus, Lactobacillus alimentarius, Lactobacillus curvatus, Lactobacillus delbruckii subsp. Lactis, Lactobacillus gasseri, Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus (Lactobacillus GG), Lactobacillus sake, Lactococcus lactis, Streptococcus thermophilus, Staphylococccus carnosus, and Staphylococcus xylosus, and mixtures thereof. More particularly, they are probiotic microorganisms derived from the group of lactic acid bacteria, especially such as Lactobacillus and/or Bifidobacterium. As illustrations of these lactic acid bacteria, mention may be made more particularly of Lactobacillus johnsonii, Lactobacillus reuteri, Lactobacillus rhamnosus, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium longum, Bifidobacterium infantis, Bifidobacterium adolescentis or Bifidobacterium pseudocatenulatum, and mixtures thereof.

A strain of Bifidobacterium lactis may be obtained from Hansen (Chr. Hansen A/S, 10-12 Boege Alle, P.O. Box 407, DK-2970 Hoersholm, Denmark) under the name Bb 12.

The species that are suitable for use are Lactobacillus johnsonii and Bifidobacterium adolescentis, which are deposited, respectively, according to the treaty of Budapest at the Institut Pasteur (28, rue du Docteur Roux, F-75024 Paris cedex 15) under the following names CNCM 1-1225 and CNCM 1-2168, and mixtures thereof.

In the case of a combination of at least one microorganism belonging to the species Lactobacillus paracasei or casei with at least one microorganism belonging to the species Bifidobacterium longum, the use or a composition may also use at least one microorganism belonging to the species Bifidobacterium lactis.

Similarly, in the case of a combination of at least one microorganism belonging to the species Lactobacillus paracasei or casei with at least one microorganism belonging to the species Bifidobacterium lactis, the use or a composition may also use at least one microorganism belonging to the species Bifidobacterium longum.

These microorganisms and/or the fractions and/or metabolites thereof may be formulated in a suitable support in an amount of at least 10³ cfu/g, in particular at doses ranging from 10⁵ to 10¹⁵ cfu/g and more particularly from 10⁷ to 10¹² cfu/g of support.

The formulations presented previously for the microorganisms belonging to the species constituting the combinations more particularly under consideration, namely Lactobacillus paracasei or casei, Bifidobacterium longum and/or Bifidobacterium lactis may, of course, be considered for the abovementioned microorganisms.

In general, the compositions may comprise for the live microorganisms from

3 15 5 15 7 12

10 to 10 cfu/g, in particular from 10 to 10 cfu/g and more particularly from 10 to 10 cfu/g of microorganisms per gram of support, or equivalent doses calculated for inactive or dead microorganisms or for microorganism fractions or for produced metabolites. The compositions for topical application generally comprise from 10³ to 10¹² cfu/g, in particular from 10⁵ to 10¹⁰ cfu/g and more particularly from 10⁷ to 10⁹ cfu/g of microorganisms, especially probiotic microorganisms.

When the composition comprises metabolites, the contents of metabolites in the compositions correspond substantially to the contents that may be produced by 10³ to

15 5 15 7 12

10 cfu, in particular 10 to 10 cfu and more particularly 10 to 10 cfu of live microorganisms per gram of support.

The microorganism(s) may be included in the composition according to the invention in a live, semi-active or inactivated, dead form.

It (they) may also be included in the form of fractions of cell components or in the form of metabolites. The microorganism(s), metabolite(s) or fraction(s) may also be introduced in the form of a freeze-dried powder, a culture supernatant and/or, where appropriate, in a concentrated form.

In the particular case of topical compositions, it may be advantageous to use these microorganisms in inactivated or even dead form.

According to one embodiment, an additional probiotic agent in accordance with the invention may be present in a content of between 0.01% and 20% by weight, preferably between 0.1% and 10% by weight and preferentially between 0.1% and 5% by weight relative to the total weight of the composition.

Besides this additional agent, the compositions according to the invention may also contain several other commonly used and/or permitted agents, these other agents being different from the active agent according to the invention.

As active agents that are conventionally used, mention may be made of vitamins B3, B5, B6, B8, C, D, E and PP, niacin, carotenoids, polyphenols, minerals and trace elements, phytoestrogens, proteins and amino acids, mono- and polysaccharides, amino sugars, phytosterols and triterpene alcohols of plant origin.

The minerals and trace elements particularly used are zinc, calcium, magnesium, copper, iron, iodine, manganese, selenium and chromium (III).

Among the polyphenols, polyphenols from grape, from tea, from olive, from cocoa, from coffee, from apple, from blueberry, from elderberry, from strawberry, from cranberry and from onion are also selected in particular. Preferably, among the phytoestrogens, isoflavones in free or glycosylated form, such as genistein, daidzein or glycitein, or lignans, in particular those from flax and from Schizandra chinensis, are selected.

A composition according to the invention may also comprise at least one antioxidant, other than the active agent according to the invention.

An additional antioxidant in accordance with the invention may be chosen from tocopherol and esters thereof, in particular tocopheryl acetate; ascorbic acid and derivatives thereof, in particular magnesium ascorbyl phosphate and ascorbyl glucoside; ferulic acid; serine; ellagic acid, polyphenols, tannins, tannic acid, epigallocatechins and natural extracts containing them, anthocyans, rosemary extracts, olive leaf extracts, for instance those from the company Silab, green tea extracts, resveratrol and derivatives thereof, ergothioneine, N-acetylcysteine, an extract of the brown alga Pelvetia caniculata, for instance Pelvetiane® from Secma, chlorogenic acid, biotin, chelating agents, such as BHT and BHA, N,N'-bis(5,5-trimethoxybenzyl)ethylenediamine and salts thereof; idebenone, plant extracts, for instance Pronalen Bioprotect TM from the company Provital; coenzyme Q10, bioflavonoids, SODs, phytanetriol, lignans, melatonin, pidolates, glutathione, caprylyl glycol, phloretin, Totarol™ or extract of Podocarpus totara containing Totarol (totara-8,11,13-trienol or 2-phenanthrenol, 4b,5,6,7,8,8a,9,10- octahydro-4b,8,8-trimethyl-l-(l-methylethyl)-; a jasmine extract such as the product sold by Silab under the name Helisun®; hesperitin laurate such as Flavagrum PEG® from the company Engelhard Lyon; an extract of Paeonia suffruticosa root, such as the product sold by the company Ichimaru Pharcos under the name Botanpi Liquid B®, an extract of lychee such as the extract of lychee pericarp sold by the company Cognis under the name Litchi derm LS 9704® and an extract of pomegranate fruit (Punica granatum), such as the product sold by the company Draco Natural Products.

An antioxidant complex comprising vitamins C and E, and at least one carotenoid, especially a carotenoid chosen from β-carotene, lycopene, astaxanthin, zeaxanthin and lutein, flavonoids such as catechins, extract of pomegranate, hesperidin, neohesperidin, proanthocyanidins and anthocyanins, may also be used.

Such antioxidants may be present in a composition in accordance with the invention in a content of between 0.01% and 20% by weight, preferably between 0.05% and 10% by weight and preferentially between 0.05% and 5% by weight relative to the total weight of the composition.

The additional agent may also be at least one prebiotic or a mixture of prebiotics. More particularly, these prebiotics may be chosen from oligosaccharides, produced from glucose, galactose, xylose, maltose, sucrose, lactose, starch, xylan, hemicellulose, inulin, gums of acacia type, for example, or a mixture thereof.

More particularly, the oligosaccharide comprises at least one fructo- oligosaccharide. More particularly, this prebiotic may comprise a mixture of fructo- oligosaccharide and of inulin.

In the topical galenical forms, it is more particularly possible to use as hydrophilic agents proteins or protein hydrolysates, amino acids, polyols, especially of C₂ to Cio, for instance glycerol, sorbitol, butylene glycol or polyethylene glycol, urea, allantoin, sugars and sugar derivatives, water-soluble vitamins, starch, and bacterial or plant extracts, for instance those from Aloe vera.

As regards the lipophilic agents, retinol (vitamin A) and derivatives thereof, tocopherol (vitamin E) and derivatives thereof, ceramides, essential oils and unsaponifiable materials (tocotrienol, sesamine, gamma-oryzanol, phytosterols, squalenes, waxes and terpenes) may be used.

It is also advantageously possible to combine the product with agents that are capable of acting:

- either directly on desquamation by promoting exfoliation, such as β-hydroxy acids, in particular salicylic acid and derivatives thereof (including 5-n-octanoylsalicylic acid); a-hydroxy acids, such as glycolic acid, citric acid, lactic acid, tartaric acid, malic acid or mandelic acid; urea and certain derivatives thereof; gentisic acid; oligofucoses; cinnamic acid; dioic acid, extract of Saphora japonica; resveratrol, detergents and certain jasmonic acid derivatives;

- and/or on the activities of enzymes involved in the degradation of comeodesmosomes, such as stratum corneum chymotryptic enzyme (SCCE), or even other proteases (trypsin-like, chymotryptic-like, cathepsin D) and also other categories of hydrolases (e.g. : glycosidases, ceramidases). Mention may be made of mineral salt chelating agents such as EDTA; N-acyl-N,N',N'-ethylenediaminetriacetic acid; aminosulfonic compounds and in particular (N-2-hydroxyethylpiperazine-N-2- ethane)sulfonic acid (HEPES); 2-oxothiazolidine-4-carboxylic acid (procysteine) derivatives; derivatives of alpha-amino acids of glycine type (as described in EP 0 852 949, and also sodium methyl glycine diacetate sold by BASF under the trade name Trilon M); honey; sugar derivatives such as O-octanoyl-6-D-maltose and N-acetylglucosamine; urea or certain derivatives thereof, for example Hydrovance; C-glycoside derivatives.

Galenical form

The compositions according to the invention may be in any galenical form normally available for the selected mode of administration, in the present case topical administration.

The support may be of diverse nature depending on the type of composition under consideration.

The compositions intended for topical administration may be aqueous, aqueous-alcoholic or oily solutions, dispersions of the solution type or dispersions of the lotion or serum type, emulsions of liquid or semi-liquid consistency of the milk type, obtained by dispersing a fatty phase in an aqueous phase (O/W) or conversely (W/O), or a suspension or emulsion of soft, semi-solid or solid consistency, of the cream or aqueous or anhydrous gel type, microemulsions, microcapsules, microparticles, or vesicular dispersions of ionic and/or nonionic type.

These compositions are prepared according to the usual methods.

They may be in the form of ointments, creams, milks, pomades, powders, impregnated pads, solutions, gels, sprays, lotions or suspensions.

These compositions may also constitute cleansing, peeling, treating or care creams for the face, the hands, the feet, the major anatomical folds or the body (for example day creams, night creams, makeup-removing creams, foundation creams or antisun creams), makeup products such as fluid foundations, makeup-removing milks, protective or care body milks, after-sun milks, skincare lotions, gels or mousses, for instance cleansing or disinfectant lotions, antisun lotions or artificial tanning lotions, bath compositions, deodorant compositions containing a bactericidal agent, aftershave gels or lotions, hair-removing creams, or compositions for combating insect stings. They may also be patches for local use.

The compositions according to the invention may also consist of solid preparations constituting soaps or cleansing bars. The topical route advantageously makes it possible to deploy all the properties of the active agent according to the invention at a precise site, on the areas to be treated.

According to one preferred embodiment, a composition in accordance with the invention is in the form of a cream, a gel, a serum, a lotion or a milk for skincare and/or for removing makeup or an aftersun composition.

When the composition of the invention is an emulsion, the proportion of the fatty phase may be between 5% and 80% by weight and preferably between 5% and 50% by weight relative to the total weight of the composition.

The oils, emulsifiers and coemulsifiers used in the composition in emulsion form are chosen from those conventionally used in cosmetics and/or dermatology. The emulsifier and the co-emulsifier may be present in the composition in a proportion ranging from 0.3% to 30% by weight and preferably from 0.5% to 20% by weight relative to the total weight of the composition.

When the composition of the invention is an oily solution or gel, the fatty phase may represent more than 90% of the total weight of the composition.

In a known manner, galenical forms intended for topical administration may also contain adjuvants that are common in the cosmetic, pharmaceutical and/or dermato logical field, such as hydrophilic or lipophilic gelling agents, hydrophilic or lipophilic agents, preserving agents, antioxidants, solvents, fragrances, fillers, screening agents, bactericides, odour absorbers and dyestuffs. The amounts of these various adjuvants are those conventionally used in the field under consideration, for example from 0.01% to 20% of the total weight of the composition. Depending on their nature, these adjuvants may be introduced into the fatty phase and/or into the aqueous phase.

As fatty substances that may be used in the invention, mention may be made of mineral oils, for instance hydrogenated polyisobutene and liquid petroleum jelly, plant oils, for instance a liquid fraction of shea butter, sunflower oil and apricot kernel oil, animal oils, for instance perhydrosqualene, synthetic oils, especially purcellin oil, isopropyl myristate and ethylhexyl palmitate, unsaturated fatty acids and fluoro oils, for instance perfiuoropoly ethers. It is also possible to use fatty alcohols, fatty acids, for instance stearic acid, and, for example, waxes, especially paraffin wax, carnauba wax and beeswax. It is also possible to use silicone compounds, for instance silicone oils and for example cyclomethicone and dimethicone, and silicone waxes, resins and gums. As emulsifiers that may be used in the invention, mention may be made, for example, of glyceryl stearate, polysorbate 60, the mixture of cetylstearyl alcohol/cetylstearyl alcohol oxyethylenated with 33 mol of ethylene oxide sold under the name Sinnowax AO® by the company Henkel, the mixture of PEG-6/PEG-32/glycol stearate sold under the name Tefose® 63 by the company Gattefosse, PPG-3 myristyl ether, silicone emulsifiers such as cetyl dimethicone copolyol, and sorbitan monostearate or tristearate, PEG-40 stearate and oxyethylenated (20 EO) sorbitan monostearate.

As solvents that may be used in the invention, mention may be made of lower alcohols, for instance ethanol, isopropanol and propylene glycol.

According to one embodiment, a composition according to the invention may comprise from 10% to 80%> by weight of water, preferably from 20%> to 70%> by weight of water and preferentially from 30% to 60% by weight of water relative to the total weight of the composition.

This water may advantageously be a spring and/or mineral water, chosen especially from Vittel water, waters from the Vichy basin and la Roche Posay water.

Hydrophilic gelling agents that may be mentioned include carboxylic polymers such as carbomer, acrylic copolymers such as acrylate/alkyl acrylate copolymers, polyacrylamides and especially the mixture of polyacrylamide, Ci₃_i₄ isoparaffin and Laureth-7 sold under the name Sepigel 305® by the company SEPPIC, polysaccharides, for instance cellulose derivatives such as hydroxyalkylcelluloses and in particular hydroxypropylcellulose and hydroxyethylcellulose, natural gums such as guar gum, locust bean gum and xanthan gum, and clays.

Lipophilic gelling agents that may be mentioned include modified clays, for instance bentones, metal salts of fatty acids, for instance aluminium stearates and hydrophobic silica, or else ethyl cellulose and polyethylene.

Cosmetic treatment process

As indicated previously, a process according to the invention may be performed by application, especially topical application, of an effective amount of at least one active agent consisting of a polypeptide having at least 80% amino acid identity with the metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1 , SEQ ID NO: 2 and SEQ ID NO: 3, or a C-terminal fragment of the said polypeptide as defined previously, for treating and/or preventing sensitive skin and the associated dysaesthetic sensations, in an individual.

According to a preferred embodiment of the invention, this process is characterized in that the effective amount of at least one active agent according to the invention is used in a composition in accordance with the invention.

According to a preferred embodiment, the process may be performed by application of creams, gels, sera, lotions or milks for removing makeup or aftersun compositions to the skin, as regards topical application.

A topical cosmetic process of the invention may be performed, for example, on a daily basis, for instance at a rate of one administration per day or one administration twice a day, for example once in the morning and once in the evening.

A cosmetic process according to the invention may be performed over a time period ranging from one week to several weeks, or even several months, this period moreover possibly being repeated after periods without treatment, for several months or even several years.

By way of example, the topical administration of a compound according to the invention may be repeated, for example two to three times daily over one day or more and generally over an extended period of at least 4 weeks, or even 4 to 15 weeks, with, where appropriate, one or more periods of stoppage.

In the description and the examples that follow, unless otherwise mentioned, the percentages are weight percentages and the ranges of values written in the form "between ... and ..." include the stated lower and upper limits.

The examples below are presented as non-limiting illustrations of the field of the invention.

EXAMPLES

1. Compositions in accordance with the invention

Example 1 : Face lotion for sensitive skin

(weight%)

Peptide derived from MMP-12 (SEQ ID NO: 4) 5.00

Antioxidant 0.05 Isopropanol 40.0 Preserving agent 0.30 Water qs 100 2. Clinical study of compositions in accordance with the invention

In order to determine the effect of a topical treatment, on the face and the legs, of a composition in accordance with the invention comprising a polypeptide derived from MMP-12 (SEQ ID NO: 4), a double- lined randomized controlled comparative single- centre study was performed on two groups of 33 individuals each.

One of these two groups used a composition A in accordance with the invention defined below, while the other group used only a placebo composition, identical to that used in the first group, except that it is free of active agent in accordance with the invention.

Composition A in accordance with the invention Oil-in-water emulsion cream

(weight%)

Parleam 5.00

Volatile silicone: glyceryl mono/distearate and polyethylene glycol stearate 15.00

Isoparaffin 5.00 Cyclopentadimethylsiloxane 15.00

Glycerol 3.00

Petroleum jelly 2.00

Peptide derived from MMP-12 (SEQ ID NO: 4) 5.00

Oil-in-water emulsion/ Arlacel/Myrj qs 100

The individuals constituting these groups each applied to their face and legs, daily for 61 days, the composition specific to the group to which they belonged.

After this period, a sensory evaluation of the efficacy of the test products on the state of skin dryness of the legs was performed by means of scores. Furthermore, a dermatologist evaluated the state of roughness of the facial skin, and a self-evaluation of the dryness was made.

In addition, the sensitivity was determined by means of a stinging test. Clinical score of the skin dryness

The investigating dermatologist evaluated at each visit every fifteen days the skin dryness of the area under study (on the outer face of the left leg) according to a scale from 0 to 3, 0 corresponding to skin that is not dry, 1 to slight dryness (slight roughness), 2 to moderate dryness (moderate roughness, a few squamae) and 3 to severe dryness (substantial roughness and desquamation).

Clinical score of the skin roughness

The investigating dermatologist evaluated by touch at each visit every fifteen days the skin roughness on both cheeks according to a scale from 0 to 4, 0 corresponding to an absence of roughness, with a perfectly smooth and supple surface.

1 corresponds to slight roughness, with slight irregularities and little roughness when touched tangentially.

2 corresponds to moderate roughness, with a markedly irregular and rough appearance and the possibility of feeling a slight induration of the skin when touched vertically.

3 corresponds to severe roughness, with a sensation of more pronounced irregularity and roughness, associated with induration of the skin.

Finally, 4 corresponds to extreme roughness, with very pronounced irregularities and major disruption of the marking of the skin, with more pronounced induration.

Self-evaluation of the state of skin dryness

The investigating dermatologist asks the volunteer at each visit every fifteen days to make a self-evaluation of the state of skin dryness on his legs and face according to a scale from 0 to 5 below: 0= not at all; 1= very slightly; 2= slightly; 3= moderately; 4= strongly; and 5= very strongly.

Stinging test

The stinging test, developed by Frosch and Kligman in 1977, makes it possible to evaluate the skin reactivity. This test determines the capacity of individuals in general to experience stinging after application of a lactic acid solution to the nasal grooves. Specifically, these areas are highly reactive and their horny layer very permeable. They are rich in hair follicles and sweat glands, which promotes the penetration of the products. Finally, they have a very dense peripheral sensory nerve network.

A dilute (10%) lactic acid solution applied to the skin immediately triggers a stinging sensation, which is proportionately more perceptible the more sensitive the skin.

By measuring the intensity of this reaction as a function of time, it is possible to assess the sensitivity of the skin.

Procedure

The stinging test is performed in the following manner:

• Application to a nasal groove of a 10% lactic acid solution and of physiological saline to the other groove: 5 to-and-fro passes (i.e. 10 passes) are performed using cotton buds soaked with each of the solutions.

30 seconds, 2 minutes and 5 minutes after the application, the volunteers evaluate the stinging sensation according to the following scale: 0: no stinging; 1 : slight stinging sensation; 2: moderate stinging sensation; and 3: severe stinging sensation.

• An overall reactivity score is calculated according to the following formula:

Overall score =∑ scores on the lactic acid side -∑ scores on the physiological saline side

The results thus obtained show that the topical treatment of the skin, and especially of sensitive skin, with an active agent in accordance with the invention leads, after two months of treatment, to an improvement in the skin barrier, which is reflected by a decrease in the dryness and sensitivity of the skin, and more generally in the reactivity of the skin versus placebo. Human metalloprotease MMP-12 in its mature form {Homo sapiens)

SEP ID NO: 1

GPVWRKHYITYRINNYTPDMNREDVDYAIRKAFQVWSNVTPLKFSKINTGMADIL

VVFARGAHGDFHAFDGKGGILAHAFGPGSGIGGDAHFDEDEFWTTHSGGTNLFLT

AVHEIGHSLGLGHSSDPKAVMFPTYKYVDINTFRLSADDIRGIQSLYGDPKENQRL

PNPDNSEPALCDPNLSFDAVTTVGNKIFFFKDRFFWLKVSERPKTSVNLISSLWPTL

PSGIEAAYEIEARNQVFLFKDDKYWLISNLRPEPNYPKSIHSFGFPNFVKKIDAAVF

NPRFYRTYFFVDNQYWRYDERRQMMDPGYPKLITKNFQGIGPKIDAVFYSKNKY

YYFFQGSNQFEYDFLLQRITKTLKSNSWFGC

Mouse metalloprotease MMP-12 (Mus musculus)

SEP ID NO: 2

RSRWMKRYLTYRIYNYTPDMKREDVDYIFQKAFQVWSDVTPLRFRKLHKDEADI

MILFAFGAHGDFNYFDGKGGTLAHAFYPGPGIQGDAHFDEAETWTKSFQGTNLFL

VAVHELGHSLGLQHSNNPKSIMYPTYRYLNPSTFRLSADDIRNIQSLYGAPVKPPSL

TKPSSPPSTFCHQSLSFDAVTTVGEKIFFFKDWFFWWKLPGSPATNITSISSIWPSIPS

GIQAAYEIESRNQLFLFKDEKYWLINNLVPEPHYPRSIYSLGFSASVK VDAAVFD

PLRQKVYFFVDKHYWRYDVRQELMDPAYPKLISTHFPGIKPKIDAVLYFKRHYYIF

QGAYQLEYDPLFRRVTKTLKSTSWFGC

Rat metalloprotease MMP-12 in its mature form (Rattus norvegicus)

SEP ID NP: 3

RSRWMKRYLTYRIYNYTPDMKRADVDYIFQKAFQVWSDVTPLRFRKIHKGEADIT

ILFAFGDHGDFYDFDGKGGTLAHAFYPGPGIQGDAHFDEAETWTKSFQGTNLFLV

AVHELGHSLGLPHSNNPKSIMYPTYRYLHPNTFRLSADDIHSIQSLYGAPVKNPSLT

NPGSPPSTVCHQSLSFDAVTTVGDKIFFFKDWFFWWRLPGSPATNITSISSMWPTIP

SGIQAAYEIGGRNQLFLFKDEKYWLINNLVPEPHYPRSIHSLGFPASVKKIDAAVF

DPLRQKVYFFVDKQYWRYDVRQELMDAAYPKLISTHFPGIRPKIDAVLYFKRHYY

IFQGAYQLEYDPLLHRVTKTLSSTSWFGC

Peptide derived from Homo sapiens MMP-12

SEP ID NP: 4

ARNQ VFLFKDDKY WLI SNLR

Peptide derived from Mus musculus MMP-12

SEP ID NP: 5

GRNQLFLFKDEKYWLINNLV Peptide derived from Rattus norvegicus MMP-12 SEP ID NO: 6

SRNQLFLFKDEKYWLINNLV

Claims

1. Cosmetic use of an effective amount of at least one active agent consisting of a polypeptide having at least 80% amino acid identity with a metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1 , SEQ ID NO: 2 and SEQ ID NO: 3 indicated below, or a C-terminal fragment of this polypeptide, for preventing and/or treating sensitive skin, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid.

2. Use according to Claim 1 , characterized in that the active agent used is a C- terminal fraction of the said polypeptide, with a length of 80 to 20 amino acids, preferably

50 to 20 amino acids and preferentially from 30 to 20 amino acids and comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid.

3. Use according to either of Claims 1 and 2, characterized in that the active agent used according to the present invention is chosen from the sequences SEQ ID NO: 4, SEQ ID NO: 5 and SEQ ID NO: 6.

4. Polypeptide with at least 80% amino acid identity with the metalloprotease MMP-12 whose sequence is chosen from SEQ ID NO: 1, SEQ ID NO: 2 and SEQ ID NO: 3, or a C-terminal fragment of the said polypeptide, for its pharmaceutical or dermato logical use for preventing and/or treating sensitive skin, the said polypeptide or the said fragment comprising the sequence KDXK, in which X represents aspartic acid or glutamic acid.

5. Cosmetic and/or dermatological composition for preventing and/or treating sensitive skin and the associated dysaesthetic sensations, comprising, in a physiologically acceptable medium, at least one active agent as defined in Claims 1 to 3, in combination with an effective amount of at least one additional probiotic agent other than the active agent as defined according to any one of Claims 1 to 3.

6. Composition according to the preceding claim, characterized in that the active agent as defined in Claims 1 to 3 is used in an amount of between 0.0001% and 30% by weight, preferably between 0.001%> and 15 > by weight and preferentially between 0.1 %> and 10% by weight, relative to the total weight of the composition.

7. Composition according to either of Claims 5 and 6, characterized in that it is intended to be used topically.

8. Composition according to any one of Claims 5 to 7, characterized in that the said additional probiotic agent is present in an amount of between 0.01% and 20% by weight, preferably between 0.1 % and 10%> by weight and preferentially between 0.1 % and 5% by weight, relative to the total weight of the composition.

9. Composition according to any one of Claims 5 to 8, characterized in that the said composition also comprises at least one fatty phase, preferably in a content of between 5%) and 80%> by weight and preferentially between 5% and 50%> by weight, relative to the total weight of the composition.

10. Composition according to any one of Claims 5 to 9, characterized in that it comprises from 10%> to 80%> by weight of water, preferably from 20%> to 70%> by weight of water and preferentially from 30% to 60% by weight of water, relative to the total weight of the composition.

11. Composition according to any one of Claims 5 to 10, characterized in that it is in the form of a cream, a gel, a serum, a lotion or a milk for skincare and/or for removing makeup or an aftersun composition.

12. Cosmetic process for treating and/or preventing sensitive skin and the associated dysaesthetic sensations in an individual, comprising at least one step of administering topically to the said individual at least an effective amount of at least one active agent as defined in any one of Claims 1 to 3.

13. Process according to Claim 12, characterized in that the said dysaesthetic sensations are chosen from stinging, tingling, itching or pruritus, burning, heating, discomfort and tautness of the skin.

14. Process according to either of Claims 12 and 13, characterized in that the said effective amount of at least one active agent as defined in any one of Claims 1 to 3 is used in a composition as defined in any one of Claims 5 to 11.