Patent Description:
It concerns the cosmetics, personal care and hygiene products and dermatology industries.

These industries are always in demand for ingredients to heal or beautify the skin and its appendages such as hair and nails, and the scalp. Moisturizing actives are among the most requested and represent very large sales volumes. They are indeed used at all ages, by any public, and at the same time for a preventive and curative purposes.

It is known that the skin tends to dry out due to environmental, psychological or hormonal factors. However, it is important that the skin be well hydrated and does not suffer loss of water that might cause and dryness. Skin dehydration is manifested by tightness, loss of flexibility and elasticity, and sometimes redness. The skin becomes rough, even squamous. Treating cutaneous dryness is an essential issue: allowing the stratum corneum to retain its flexibility and its barrier function, is also a more global way of protecting the underlying dermis and preventing skin aging.

A dry skin profile, if coupled with a skin abnormality of clogged pores, can lead to a benign condition, the keratosis pilaris.

Pilaris keratosis, quite common in men and women, results from hyperkeratosis. On certain areas of the body such as the legs, upper arms, waist and sometimes cheeks or buttocks, the skin is covered with small unsightly blisters, clearly visible, palpable, often without itching and not necessarily erythematous, which make it appear like a "goose bumps".

This mild skin disorder affects <NUM>% of the world's female population of all ages, although more commonly seen at puberty and during pregnancy. It is characterized by an accumulation of corneocytes too abundantly produced in and at the exit of the hair follicle forming a keratinized plug which blocks and then dilates the hair follicle.

It is favored and amplified by a dry environment. Thus, keratosis pilaris is also more common in people with dry skin (ichthyosis vulgaris and atopic dermatitis), with a less effective skin barrier, less retention of epidermal water.

It is therefore a problem of keratinization of dry skin. The problem is solved by improving the process of maturation of the epidermis that restores its homeostasis. The keratinocytes of the upper part of the epidermis should be conducted to transform themselves more effectively into corneocytes to form an effective skin barrier. This makes it possible to regulate the production of moisturizing elements and lipids of the stratum corneum, to improve osmolyte flow, to maintain good skin hydration and thus to reduce the effects of keratosis pilaris.

The epidermal floor, called the basal layer, continuously produces new keratin-rich cells, called keratinocytes, that migrate to the outside of our body and undergo a major transformation throughout this migration. Keratinocytes arrive at the granular layer and produce structuring or moisturizing protein granules and precursor lipids for ceramides used to seal the skin or control its hydration. The keratinocytes produce and bind cornification proteins, such as involucrine and loricrin, with transglutaminase. They also manufacture moisturizing compounds from filaggrin, then they lose their nucleus and release the lipids between the cells. Loricrin and filaggrin are strongly diminished in keratinocytes deficient in the CALML5 protein thus indicating the crucial role of this protein. Then the corneocytes, strongly attached to each other by protein bonds, are detached from each other and produce the scales.

Thus, normal skin provides a protective function with respect to transepidermal water loss (TEWL). Surface cutaneous lipids in humans are a bulwark between the body and the environment. Any quantitative or qualitative abnormality of these lipids leads to an increase in the insensitive water loss (TEWL), a marker of the alteration of the barrier function.

Therefore, the barrier function is not complete without the indispensable contribution of various lipids, perfectly architected between them, which are produced by the keratinocytes of the granular layer, whose granules carry the protein ABCA12 (or ABC transporter A12). The latter fills its granules with ceramide precursors, allows their transport and their deposition between the keratinocytes of the stratum corneum, thanks to the capture and hydrolysis of ATP. Ceramides represent <NUM>% of the lipids of the barrier. Mutations of ABCA12, by reducing ceramide intakes by up to <NUM>%, lead to disorders of keratinization and alterations of the barrier function by the disorganization of the lipid architecture of the stratum corneum.

Water gradient drops rapidly in the epidermis from <NUM>% in the spinous layer to <NUM>% at the granular layer-stratum corneum interface and <NUM>% at the surface. At the granular level, fluctuations in ambient humidity greatly modulate the proteolysis of filaggrin in its moisturizing by-products, which can lead to its collapse, because the enzymes in this process require an optimum of intracellular hydration.

To continue to ensure their role in the survival of the individual and reduce water loss, keratinocytes are equipped with sensors and osmolarity control systems consisting of membrane channels regulating the molecular exchanges between the inside and the outside of the cell. TAUT, one of these channels, allows the spinous and especially granular cells, the most exposed to dehydration, to concentrate taurine, a water retaining osmolyte. TAUT expression is stimulated by keratinocyte maturation and osmotic shock. Myoinositol is also an osmolyte of interest for the epidermis; its flows are managed by SMIT1 (sodium-myoinositol cotransporter-<NUM>). Like TAUT, SMIT is induced in the human keratinocyte by hyperosmosis which causes the entry of their respective osmolytes.

Aquaporin-<NUM> (AQP3) is another of these carrier channels; it is the aquaporin most expressed by the viable epidermis, including granular layer. This membrane protein is part of a family that allows the movement of water in the cell, but AQP3 also controls the transmembrane flows of glycerol, an osmolyte known for its moisturizing properties in the epidermis. Mice deficient in AQP-<NUM> have less water in their stratum corneum. It was also observed a delay in syntheses of lipids of the stratum corneum, an increase of the TWEL and a delay of reconstitution of the barrier after abrasion.

Furthermore, one of the important compounds for maintaining hydration of the epidermis is a very large negatively charged carbohydrate polymer: hyaluronic acid (or hyaluronan or hyaluronate). It is produced by keratinocytes, in smaller quantities compared to dermis, and is positioned around each cell where it plays a role in the solvation of vitamins and electrolytes. Hyaluronic acid ensures the capture of a very large number of water molecules: up to <NUM> times its own weight. This allows the transfer of vitamins and solutes because the epidermis is not vascularized.

<CIT> disclosed an extract of Salinicoccus hispanicus culture from inactivated cells or bacterial cell membranes.

The aim of the present invention is to propose an active agent, more particularly a moisturizer, which will act in the most complete possible way on the various biological mechanisms described above involved in the treatment of dry skin, such as the sensible water loss, the reinforcement of the cutaneous barrier, and the osmolytes responsible for hydration.

Another aim of the invention is to provide an active having a more specific action on keratosis pilaris, the cutaneous dryness constituting a favorable ground for the development of this unsightly skin disorder.

To this aim, the present invention provides a cosmetic or dermatological active ingredient comprising a bacterial extract according to claim <NUM>. Preferably this extract is an aqueous extract.

By intracellular extract is meant the bacterial fraction soluble in the external liquid medium, obtained after the lysis of the bacteria and the separation by centrifugation of soluble and insoluble fractions (membranes, debris.

The bacterium Salinicoccus hispanicus lives in moderately saline environments. It belongs to the phylum of firmicutes that have a strong ability to adapt to changes in their environment. Salinicoccus hispanicus for example survives in a highly desiccant and saline environment.

Surprisingly, the Applicant has discovered that Salinicoccus hispanicus in culture is able to withstand salt stress by producing an original set of molecules that can be used in cosmetics and dermatology to treat, improve or embellish the condition of the skin, its appendages (body hair, hair, nails, etc.), and scalp.

Another advantage of this bacterium is its low impact on the environment, since it only uses water and natural substrates for its growth and its cultivation is done at room temperature, which limits the consumption of energy.

The intracellular extract of Salinicoccus hispanicus according to the invention is characterized by the composition of its dry matter mainly containing proteins (amino acids to proteins), significant amounts of mineral salts and nucleotides, and traces of sugars and lipids.

The bacterial extract according to the invention contains:.

The subject matter of the invention consists of a cosmetic or dermatological active ingredient containing the intracellular extract of Salinicoccus hispanicus according to the invention defined above and a physiologically acceptable medium. Preferably, the physiologically acceptable medium is hydrophilic in nature.

Another subject matter of the invention is a cosmetic or dermatological composition according to claim <NUM>, and optionally one or more additional active ingredients, in a physiologically acceptable medium. The additional active agents are preferably chosen from: compounds of vitamin B3, niacinamide, tocopherol, retinol, hexamidine, α-lipoic acid, resveratrol, DHEA, hyaluronic acid, a moisturizing and/or emollient product, and one or more peptides, which are active ingredients usually used in cosmetics.

The moisturizing and/or emollient product may be chosen from plant extracts and vegetable butters/vegetable oils, fatty acid esters, squalanes and ceramides, extracts of animal origin, sugar derivatives, probiotics and prebiotics, alpha-glucans, oligosaccharides and glycols.

In the composition according to the invention, the active ingredient may be in a free form and optionally in a bound form, incorporated or adsorbed on macro-, micro-, and nanoparticles, or on macro-, micro- and nanocapsules, for the treatment of textiles, natural or synthetic fibers, wool, and any material intended to come into contact with the skin, such as, for example, day or night underwear, wipes, tissues, or fabrics.

Afurther subject matter of the invention relates to the use according to claim <NUM>.

In vitro and in vivo tests described below show that the bacterial extract according to the invention is particularly effective for treatment of skin (including the scalp) which is dry or exhibits keratosis pilaris, in particular through smoothing and/or hydration of the skin and/or scalp. The skin is visibly more beautiful. Its grain is refined, more homogeneous thanks to a smoothing effect of the micro-relief. Feelings of tightness, discomfort, and redness that may or may not be associated with dry skin, are also eliminated and skin aging that is too rapid is avoided through hydration and strengthening of the dermal-epidermal junction (DEJ). The extract according to the invention may for example be used for a moisturizing treatment post-depilatory or post-peeling. According to the invention, this treatment involves in particular reinforcing the function of the cutaneous barrier and/or the regulation of the ionic fluxes in the cells of the skin and/or the protection of these cells with respect to the osmotic shocks and/or stimulation of the synthesis of laminin <NUM> and collagen <NUM>, two of the major proteins of DEJ. DEJ separates the dermis from the epidermis and plays an important role in the organization and structuring of the epidermis and therefore in the surface state of the skin. In addition to its role as a support for anchoring the epidermis to the dermis, DEJ determines the adhesion of basal keratinocytes to DEJ proteins in a well-defined orientation. It also ensures transmission of proliferation and differentiation signals from the dermis to the epidermis.

A disorganization of the DEJ therefore has adverse consequences at the epidermal level and impacts the barrier function. It is also considered as one of the causes of skin aging.

Preferably, the active ingredient is used at a concentration of <NUM>% to <NUM>% by weight relative to the total weight of the composition, more preferably at a concentration of <NUM> to <NUM>%.

The active ingredient according to the invention or said composition according to the invention is disclosed for the treatment of a disease of the skin and/or scalp. Preferably, said treatment is carried out topically.

The in vitro and in vivo tests detailed below in the description demonstrate the beneficial effects of the cosmetic treatment according to the invention, in particular on keratosis pilaris:.

"Physiologically acceptable medium" means according to the present invention, without limitation, an aqueous or aqueous-alcoholic solution, a water-in-oil emulsion, an oil-in-water emulsion, a microemulsion, 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 use in contact with the skin and scalp of mammals and more particularly of human, without risk of toxicity, incompatibility, instability, allergic response, among others. This "physiologically acceptable medium" forms what is usually called the excipient of the composition.

In a composition according to the invention, the bacterial extract may be combined with other active ingredients at effective concentrations that can act synergistically or in a reinforcing manner to achieve the desired effects described for the invention, such as the following agents: filtering radiation, in particular UVA and/or UVB, moisturizing, humectant, calming, myorelaxing, slimming, restructuring, firming, plumping, tensing, anti-dandruff, acting on the microcirculation, acting on inflammation, on free radicals, anti-wrinkles, lightening, acting on the radiance of the complexion, anti-glycation, propigmenting, acting on the stratum corneum, on the dermis-epidermis junction, on the production of protein HSPs, on the firmness, the elasticity, the tonicity of skin, regrowth of hair, peptides, vitamins etc..

The treatment according to the invention can be applied to all parts of the body, and more specifically according to the indication recommended on the face, body, neckline or scalp, in any form or vehicle known to the skilled persons in the art, particularly in the form of a solution, dispersion, emulsion, paste or powder, individually or in premix or be conveyed individually or premixed with vectors such as macrocapsules, microcapsules or nanocapsules, macrospheres, microspheres, or nanospheres, liposomes, oleosomes or chylomicrons, macroparticles, microparticles or nanoparticles, macro-sponges, micro-sponges or nanosponges, microemulsions or nanoemulsions, or adsorbed on polymers powdery organic, talcs, bentonites, spores or exines and other mineral or organic carriers.

In cosmetics in particular, applications can be proposed especially in the skincare ranges of the face, body and scalp.

CTFA ("<NPL>) describes a wide variety, without limitation, of cosmetic ingredients usually used in the skincare and scalp care industry, which are suitable for use as additional ingredients in the compositions of the present invention.

Other additional skin care actives that are particularly useful can be found in Sederma's commercial literature and at www.

The following commercial actives can also be mentioned as examples: betain, glycerol, Actimoist Bio <NUM>™ (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™ (nom commercial de l'acétyl hexapeptide-<NUM> from Lipotec), spilanthol or an extract of Acmella oleracea known under the trade name Gatuline Expression™, an extract of Boswellia serrata known under the name Boswellin™, Deepaline PVB™ (Seppic), Syn-AKE™ (Pentaphann), Ameliox™, Bioxilift™ (Silab), PhytoCellTec™Argan (Mibelle), Papilactyl D™ (Silab), Preventhelia™ (Lipotec), or one or more of the following active ingredient marketed by Sederma : Subliskin™, Venuceane™, Moist <NUM>™, Vegesome Moist <NUM>™, Essenskin™, Juvinity™, Revidrat™, Resistem™, Chronodyn™, Kombuchka™, Chromocare™, Calmosensine™, Glycokin factor S™, Biobustyl™, Idealift™, Ceramide <NUM>™, Ceramide A2™, Ceramide HO3™, Legance™, Intenslim™, Prodizia™, Beautifeye™, PacifeelTM, Zingerslim™, Meiritage™, Senestem™, Sebuless™, Majestem™, Apiscalp™, Rubistem™, CitystemTM, Neonyca™, NG Insaponifiables de Beurre de Karité™, Majestem™, or mixture thereof.

Among plant extracts (in the form of conventional extracts or prepared by an in vitro method) that can be combined with the bacterial extract according to 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 artichoke (Cynara scolymus), 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. 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 Globularia cordifolia, of Albizzia julibrissin, of Oxydendron arboretum, of Zingimber zerumbet smith, of Astragalus membranaceus, of Atractylodes macrocephalae, of Plantago lanceolata, of Leontopodium alpinum, of Mirabilis jalapa, of Marrubium vulgare, or of orchids.

The compositions of the present invention may include one or more peptides, including, without limitation, di-, tri-, tetra-, penta-and hexapeptides and their derivatives. According to a particular embodiment, the concentration of the additional peptide, in the composition, ranges from 1x10-<NUM>% and <NUM>%, preferably from 1x10-<NUM>% and <NUM>%, preferably between 1x10-<NUM>% and <NUM>% by weight.

According to the present invention, the term "peptide" refers to peptides containing <NUM> 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 (βAH), YR, VW, NF, DF, KT, KC, CK, KP, KK, TT, PA, PM or PP.

Suitable tripeptides for use herein include, but are not limited to RKR, HGG, GKH, GHK, GGH, GHG, KFK, KAvaK, KβAK, KAbuK, KAcaK, KPK, KMOK, KMO<NUM>K (MO<NUM> being a di-oxygenated sulfoxide methionine), KVK, PPL, PPR, SPR, QPA, LPA or SPA.

Suitable tetrapeptides for use herein include but are not limited to RSRK (SEQ ID NO: <NUM>), GQPR (SEQ ID NO: <NUM>), KTFK (SEQ ID NO: <NUM>), KTAK (SEQ ID NO: <NUM>), KAYK (SEQ ID NO: <NUM>) or KFYK (SEQ ID NO: <NUM>).

Suitable pentapeptides include but are not limited to KTTKS (SEQ ID NO: <NUM>). Suitable hexapeptides include but are not limited to GKTTKS (SEQ ID NO: <NUM>) and VGVAPG (SEQ ID NO: <NUM>).

Other suitable peptides for use herein include but are not limited to: lipophilic derivatives of peptides, preferably palmitoyl (Pal) derivatives, and metal complexes as aforementioned (e.g. copper complex of the tripeptide HGG). Preferred dipeptides include for example N-Palmitoyl-β-Ala-His, N-Acetyl-Tyr-Arg-hexadecylester (Calmosensine™, Idealift™ from Sederma), Pal-RT or Pal-KT (Sederma). Preferred tripeptide derivatives include for example 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<NUM> (Peptide CK+), N-Biot-GHK (from Sederma), Pal-KAvaK, Pal-KβAlaK, Pal-KAbuK, Pal-KAcaK, or Pal-KMO<NUM>K (Matrixyl®synthe'<NUM>® from Sederma), PalKVK (Syn-Coll™ of DSM), and derivatives thereof.

Here can further be cited also the propigmenting and/or pro-mec dipeptides and tripeptides of general formula X-(Xaa<NUM>)n-Pro*-Xaa<NUM>-Y disclosed in <CIT>, with n=<NUM>, <NUM> or <NUM>, Xaa<NUM> an hydrophobic aminoacid selected from Ala, Val, Met, Leu, Iso, Phe, Pro, and analogs and derivatives thereof; or a polar aminoacid selected from Ser, Thr, Tyr, Asp, Glu and analogs and derivatives thereof; and when n=<NUM> the two aminoacids Xaa<NUM> being the same or different; Xaa<NUM> being an hydrophobic aminoacid selected from Ala, Val, Met, Leu, Iso, Phe, and analogs and derivatives thereof, or a basic aminoacid selected from Arg, Lys, His, and analogs and derivatives thereof; at the N terminal end X being selected from H, -CO-R<NUM> and -SO<NUM>-R<NUM>; at the C terminal end Y being selected from OH, OR<NUM>, NH<NUM>, NHR<NUM> or NR<NUM>R<NUM>; R<NUM> and R<NUM> being, independently from each other, selected from an alkyl, aryl, aralkyl, alkylaryl, alkoxy et aryloxy group, that can be linear, branched, cyclic polycyclic, saturated, unsaturated, hydroxylated, carbonylated, phosphorylated and/or sulfured, said group having or not an O, S and/or N heteroatom in its skeleton and Pro* corresponding to a Proline, analog or derivative thereof; comprising for example the following peptides Pal-SPR-OH, Pal-PPR-OH, Pal-QPA-OH, Pal-LPAOH, Myr-SPA-OH, Pal-PM-OH, Pal-PA-OH and Pal-PP-OH.

Suitable tetrapeptide derivatives for use according to the present invention include, but are not limited to, Pal-GQPR (SEQ ID NO: <NUM>) (from Sederma) and Pal-KTFK (SEQ ID NO: <NUM>) or Ela-KTFK (SEQ ID NO: <NUM>), Ela-KTAK (SEQ ID NO: <NUM>), Ela-KAYK (SEQ ID NO: <NUM>) or Ela-KFYK (SEQ ID NO: <NUM>). Suitable pentapeptide derivatives for use herein include, but are not limited to, Pal-KTTKS (SEQ ID NO: <NUM>) (available as Matrixyl® from Sederma), Pal-YGGFXaa (SEQ ID NO: <NUM>) with Xaa being Leu or Pro, or mixtures thereof. Suitable hexapeptide derivatives for use herein include, but are not limited to, Pal-VGVAPG (SEQ ID NO: <NUM>), Pal-GKTTKS (SEQ ID NO: <NUM>), Pal-HLDIIXaa with Xaa being Trp, Phe, Tyr, Tic, <NUM>-hydroxy-Tic ou Tpi (SEQ ID NO: <NUM>) and derivatives thereof. The mixture of Pal-GHK and Pal-GQPR (SEQ ID NO: <NUM>) (Matrixyl® <NUM>, 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'<NUM>® of Sederma. The compositions commercially available preferred sources of tetrapeptides include Rigin™, Eyeliss™, Matrixyl® Reloaded and Matrixyl <NUM>® which contain between <NUM> and <NUM> ppm of Pal-GQPR (SEQ ID NO: <NUM>) and an excipient, proposed by Sederma.

The following marketed peptides can be mentioned as well as additional active ingredients:.

More specifically, according to the invention the alkyl-phthalide(s) or plant extract comprising such may be combined with at least one of compounds selected from compounds of the vitamin B3, compounds such as niacinamide or tocopherol, retinoid compounds such as retinol, hexamidine, α-lipoic acid, resveratrol or DHEA, hyaluronic acid, peptides, in particular N-acetyl-Tyr-Arg-O-hexadecyl ester, Pal-VGVAPG (SEQ ID NO: <NUM>), Pal-KTTKS (SEQ ID NO: <NUM>), Pal-GHK, Pal-KMO<NUM>K and Pal-GQPR (SEQ ID NO: <NUM>), which are widely used active ingredients in topical cosmetic or dermopharmaceutical compositions.

More specifically, the bacterial extract according to the invention can be combined with a moisturizing and/or emollient product for the skin, its appendages (nails, eyelashes, eyebrows), the scalp, the hair, in particular one or more compounds below:.

The present invention also provides a cosmetic topical treatment method for improving the appearance and general condition of the skin and scalp, including topical application to the skin of a subject in need thereof of an effective amount of at least one bacterial extract according to the invention or a composition comprising it, in a physiologically acceptable excipient.

By "topical treatment" or "topical use" is meant an application that is intended to act at the place where it is applied: skin or scalp.

A composition according to the invention can be applied locally to the targeted areas, for example using a canula-type applicator suitable for the scalp.

The "effective" amount depends on a variety of factors, such as age, condition of the patient, severity of the disorder or condition and the way of administration. An effective amount means a non-toxic amount sufficient to achieve the desired effect.

All percentages and ratios used in this application are by weight of the total composition and all measurements are made at <NUM> unless otherwise specified.

For example, for a cosmetic facial treatment, the European Cosmetics Directive has set a standard application amount of a cream of <NUM>/cm<NUM>/day/person and for a lotion for the body of <NUM>/cm<NUM>/day/person.

According to other features, the cosmetic treatment method according to the invention may be associated with one or more other treatment methods for the skin, such as, for example, light therapy, heat or aromatherapy treatments.

According to the invention, it is possible to propose devices with several compartments or kits intended for the implementation of the method described above, and which could include, by way of example, and without being limiting, in a first compartment a composition containing at least the bacterial extract and in a second compartment an additional excipient and/or active, the compositions contained in said first and second compartments being here considered as a combination composition for simultaneous, separate or spread over time use, especially in one of the treatments defined above.

The treatment method according to the invention is more particularly suitable for a hydration treatment, smoothing, soothing, a treatment for reducing redness and feelings of tightness and cutaneous discomfort, especially post-depilation or combined with an exfoliating or post exfoliation treatment, and/or a treatment of keratosis pilaris, and/or a treatment intended to slow down skin aging.

The present invention will be better understood in light of the following detailed description and illustration examples.

The Salinicoccus hispanicus strain is cultured in a fermentation medium comprising in particular sugars, proteins (peptone, yeast extract) and highly enriched in salts (especially NaCl). The salt content can range from <NUM> to <NUM>/l.

The temperature of the culture may be between <NUM> and <NUM>.

Additions of compounds are possible at different times in the culture.

The culture is stopped when the OD (optical density) at <NUM> reaches between <NUM> and <NUM> (measurement of turbidity).

The assembly bacteria + medium is then centrifuged, the bacteria washed with water and the fresh biomass recovered in <NUM>% of the initial total volume (concentration of the biomass by a factor of <NUM>). The bacteria are then lysed either mechanically or chemically. The intracellular content is thus released into the external environment. Envelopes and cell debris are removed by centrifugation. The pH of the supernatant is adjusted to <NUM>. The supernatant is filtered.

The filtrate constitutes said "extract according to the invention" used hereinafter.

This bacterial extract can be characterized by the composition of its dry matter:.

The active ingredient comprises the extract according to the invention obtained according to A) above pure or diluted in a matrix forming a physiologically acceptable medium. This active ingredient is especially intended for the cosmetic industry for the preparation of cosmetic products, creams, gels, etc..

(see galenic examples in point D) below). The active ingredient will preferably be formulated between <NUM> and <NUM>% in a cosmetic composition applicable to the skin, preferably at a few%, especially <NUM> to <NUM>%, depending on the more or less concentrated effect desired.

The dilution of the extract according to the invention can be carried out in any physiologically acceptable hydrophilic excipient. Water and water-miscible solvents of the type, for example glycerin or any other physiologically acceptable short chain polyol, will preferably be used.

For example, for the formulas of the galenic of point D), which will be used for the in vivo tests, it is a dilution by a factor <NUM> which has been preferentially used to manufacture the active ingredients.

For the in vitro tests, the pure extract according to the invention will be used in the appropriate aqueous medium of each test.

Various cosmetic formulations are described below. Additional active ingredients, acting in support and/or in addition to the activity of the active ingredient according to the invention may be added in the appropriate phase according to their hydrophobic or hydrophilic nature. These ingredients can be of any category according to their function(s), the place of application (scalp, face, body, neck, bust, hands, etc.), the desired end effect and the targeted consumer.

The active ingredient according to the invention used in the galenical formulations given below is the extract according to the invention diluted by a factor of <NUM> in an aqueous excipient.

For the in vitro tests on skin explants and in vivo tests, the cream <NUM>) described below was used.

A first series of tests focused on the reinforcement and maturation of the barrier through the stimulation of several markers of keratinocyte differentiation.

A second series of tests focused on improving the hydration of the epidermis through the stimulation of hyaluronic acid and osmolytes transporters.

A third series of tests focused on strengthening the JDE, through the stimulation of two of its main structural proteins: collagen <NUM> and laminin <NUM>.

A final test has shown the interest of the product to stimulate the production of ATP and thus to bring energy to the skin cells.

The in vitro tests below were performed on several different biological models all of human origin: keratinocytes and skin explants from cosmetic surgery. The extract according to the invention, as prepared in point A) above, was tested either directly at the indicated concentrations (for cell culture tests) or integrated into the cream <NUM>) described above in the Galenic part C) (for explant tests).

Normal human keratinocytes almost at confluence are brought into contact with the extract according to the invention (between <NUM> and <NUM>%) in a suitable culture medium in order to study their differentiation under a microscope by following the appearance of the particular layers using varied and appropriate microscopic techniques.

No hyperproliferation of KH was observed with the extract according to the invention. With the extract according to the invention, a clear acceleration of the differentiation with respect to the control is observed. This results in the rapid onset (<NUM> days) of typical structures of the upper layers of the epidermis (presence of branched structures characteristic of the rigid proteolipid matrix and in
multilayer of the horny envelope; characteristic intracellular network). At the same time of observation, the control mat does not have such a differentiated profile.

Thus, the bacterial extract according to the invention is able to stimulate the differentiation of KH in order to establish an effective barrier.

Dry skins, and in particular those of persons prone to keratosis pilaris, are characterized by deficient levels of intercellular lipids at the level of the stratum corneum (including in particular ceramides and neutral lipids). This contributes to the weakening of the cutaneous barrier. In addition, involucrin, loricrin, filaggrin, transglutaminase and CALML5 are among the proteins responsible for the good quality of the barrier function of the stratum corneum, allowing "normal" skin to limit water losses at level of the barrier.

Therefore, in order to improve dry skin, cosmetic active agents capable of stimulating the production and/or the expression of these proteins and/or the production of these lipids will be sought.

On the same mats as those used above to observe differentiation, a number of differentiation markers were evaluated.

The same culture and contact protocol as previously was used on human keratinocytes. The keratinocytes were then labeled by immunocytochemistry using antibodies specific to markers of cornification/maturation of these cells: involucrin, loricrin, filaggrin and ceramide-<NUM>. Reproducible photos were made and quantified by image analysis, in comparison with the control.

Moreover, the increase in the expression of transglutaminase-<NUM> but also the transglutaminase activity and the amount of neutral lipids produced by KH in culture in contact with the extract according to the invention were measured and compared with those of their control without product. The methods were respectively: qRT-PCR, enzyme-substrate enzymatic activity assay and red oil labeling of neutral lipids included in KH.

Finally, the cream <NUM>) described above in Galenic part C) was applied to skin explants (female, <NUM> years old, abdomen) which were then sectioned to be immunohistologically labeled to evaluate CALML5. The photos taken were then analyzed by image analysis and compared to placebo cases.

The results show the stimulating effect of the bacterial extract according to the invention on the maturation of KH in culture. There is a higher expression of the protein involucrine in KH (+<NUM>%; p<<NUM>), as well as loricrin protein (+<NUM>%; p<<NUM>) which are respectively weakly expressed or almost absent in their case control.

Table <NUM>: Variation of the expression of filaggrin and ceramide-<NUM> by KH in the presence of the bacterial extract according to the invention; Immunocytochemistry, n=<NUM>.

The results confirm the stimulatory effect of the bacterial extract on the maturation of KH in culture. Thus, a higher expression of the filaggrin protein in KH (+<NUM>%, p<<NUM>) and ceramide-<NUM> lipid (+<NUM>%, p <<NUM>) is also observed, both being almost absent in control cases.

The results show a higher production of neutral lipids in KH (x <NUM>; p<<NUM>), even though they are almost absent in the control case.

These results show that the bacterial extract according to the invention effectively promotes the expression of transglutaminase-<NUM> mRNAs (+ <NUM>%; p<<NUM>). The complementary study on the intracellular activity of transglutaminases shows an increase (+ <NUM>%; p <<NUM>) which confirms the effect of the bacterial extract according to the invention on these cells.

The cream <NUM>) described above in Galenic part C), applied for <NUM> consecutive days on <NUM> skins, increased + <NUM>% the signal corresponding to CALML5 compared to the placebo cream applied on an equivalent number of control skins. This increase is significant at-risk p<<NUM>.

In conclusion, it appears that the extract according to the invention stimulates all the tested markers of the maturation of the stratum corneum, with a view to reinforcing the skin barrier effect.

The beneficial effects of the bacterial extract according to the invention on the regulation of hydration have been studied with several models.

Firstly, skin explants (woman, <NUM> years, abdomen): the two creams, the one containing the extract according to the invention (cream <NUM>) described above in Galenic part C), the other its placebo, were applied to the stratum corneum daily for <NUM> days. At the end of this step, the skins were cut into thin sections (<NUM>) and the DSG-<NUM> protein was labeled by immunohistology. On cuts from the same trial, AQP-<NUM>, a major water and glycerol transporter, was also immunohistologically labeled and quantified by image analysis on photos. The markings obtained with the cream containing the extract according to the invention were compared with those obtained with the placebo cream.

In a second series of tests, the extract according to the invention, in solution in the culture medium, was brought into contact with human keratinocytes (KH) and then a hyaluronic acid assay was performed by a method derivative from Elisa on culture media.

In a third series of tests, the ability of the extract according to the invention to promote mRNA production of two other cell-important osmolyte transporters, TAUT and SMIT-<NUM>, was assessed by qRT-PCR. After contact with the extract according to the invention, human keratinocytes were crushed and mRNAs corresponding to these proteins were evaluated by qRT-PCR after conversion into cDNA.

In the same way, the expression of the mRNAs of the ABCA12 protein was evaluated.

These results show that the extract according to the invention positively modulates two proteins important for the water homeostasis of the skin: DSG1 and AQP3. The first to maintain cell cohesion between them and the second to manage the flow of electrolytes. There was an increase of +<NUM>% (p<<NUM>) for DSG-<NUM> and +<NUM>% (p <<NUM>) for AQP3.

These results show that the extract according to the invention positively modulates the production of hyaluronic acid which is known to be deposited around these cells forming a kind of hydric bubble. The latter allows the transit of osmolytes, nutrients and vitamins. The effect is clear from <NUM>% and reaches +<NUM>% (p<<NUM>).

As previously, a very positive effect of the extract according to the invention for modulating the TAUT and SMIT1 proteins, known to regulate the flow of epidermal electrolytes, is noticed. TAUT mRNA expression increase was +<NUM>% (p<<NUM>) and that of SMIT1 +<NUM>% (p<<NUM>).

These results show that ABCA12 is increased early under the influence of the extract according to the invention: + <NUM>% (<NUM>, p<<NUM>) and + <NUM>% (<NUM>, p<<NUM>).

In conclusion, it appears that the extract according to the invention stimulates all the markers going in the sense of a reinforced hydration of the skin.

Laminin <NUM> is important at the level of the DEJ. It ensures the proper anchoring of basal keratinocytes on the basement membrane and is responsible for the suppleness of the epidermis. In addition, it stimulates the proliferation of keratinocytes, allowing them to engage in differentiation. On older cells Laminin <NUM> is no longer replaced as efficiently as on young cells, hence the interest of stimulating the biosynthesis for a better renewal.

The type VII collagen is in the form of fibrils and allows the anchorage of the DEJ to the elastic fibers of the dermis.

In a first series of tests, the extract according to the invention in solution in the culture medium was brought into contact with human keratinocytes. An assay of laminins and collagen VII was made on the media; the result is reduced to the number of cells present on the layers.

In a second series of tests laminin <NUM> was evaluated on skin explants. The two creams, the one containing the extract according to the invention: cream <NUM> described above in Galenic part C), the other its placebo, were applied to the stratum corneum every day for <NUM> days. At the end of this step, the skins were cut into thin sections (<NUM>) and the laminin <NUM> was labeled by immunohistology and then quantified by image analysis in photos. The labelings obtained with the cream containing the extract according to the invention were compared with those obtained with the placebo cream.

The extract according to the invention positively modulates the production of laminins. The effect is clear from <NUM>% and reaches +<NUM>% (p<<NUM>).

The extract according to the invention positively modulates the production of collagen VII. The effect is clear from <NUM>% and reaches +<NUM>% (p<<NUM>).

These results show that the extract according to the invention positively modulates laminin <NUM> synthesis. An increase of +<NUM>% is noted (p<<NUM>).

The extract according to the invention stimulates the synthesis of laminin and collagen VII favoring better anchoring of basal keratinocytes to JDE, their proliferation/differentiation and a better anchoring of the DEJ on the elastic components of the dermis.

In preliminary studies on human keratinocytes, it has been shown that <NUM> and <NUM>% of the extract according to the invention strongly stimulate the neo-synthesis of ATP in these cells by respectively +<NUM>% and +<NUM>% (in both cases with p<<NUM>).

Thanks to this action, the extract according to the invention allows the cells of the skin to have energy enabling them to meet at least part of their needs.

An evaluation of the skin efficacy on the smoothing, hydration, restructuring, soothing, reduction of discomfort and post-stress redness of the active ingredient according to the invention used at <NUM>% was conducted out of a total of <NUM> volunteers in <NUM> independent studies:.

This first study was carried out on a total panel of <NUM> women of <NUM> years old age (<NUM> - <NUM> years), having the skin of legs dry with visible scales and feeling systematically discomfort following a depilation of the legs with wax. In addition, many of them (N = <NUM>) had keratosis pilaris on the upper arms or thighs. Prerequisite conditions and/or during the study were imposed on the volunteers (absence of shaving, use of creams on the legs, exposure to the sun, etc.) in order to limit artifactual effects.

This study was conducted in single blind on the legs and upper arms. The volunteers applied a cream containing <NUM>% of the ingredient according to the invention (cream according to the invention corresponding of formula <NUM>) of point D) above) and a placebo cream (without active ingredient) in contra-lateral. The creams were applied in bi-daily massage for <NUM> weeks.

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

Statistical studies were performed using Student's t-test or if needed with a non-parametric Wilcoxon test. Bilateral tests were performed on matched series. For questionnaire evaluations, a Khi2 test was used.

Standardized photos were captured directly on a dry area of the lower leg using a dermoscopic camera using constant LED illumination and a high precision optical system to perfectly standardize colors, brightness and geometry.

At T0, T1week and T4 weeks, <NUM> acquisitions were made on <NUM> neighboring but independent sites. From the images obtained, a selective extraction of the scales was carried out using a software and a percentage of squams was obtained for each case.

The application of the cream according to the invention leads to a significant improvement, significant from <NUM> week, with a decrease in the amount of scales of <NUM>% (p<<NUM> vs. T0 and p<<NUM> vs. placebo), while the application of a placebo leads to a lower decrease of <NUM>%.

After <NUM> weeks of application of the products, a cumulative effect causes almost disappearance of the scales with -<NUM>% for the cream according to the invention and <NUM>% for the placebo (p<<NUM> for both vs. T0). The difference in effect remains significant in favor of the cream according to the invention (p<<NUM>).

The standardized photos, previously captured, were appreciated by a panel of <NUM> expert judges. For each volunteer, the experts visualized the pictures before and after treatment (at T0, T1sem and T4sem. ) And quantified the attenuation of scales according to <NUM> grades. For the presentation of the results, a binary grouping was carried out to separate the "significant" effects from the "negligible" effects.

The results show that the experts see a noticeable decrease in scales from the first week of application of the cream according to the invention with an occurrence of <NUM>%. This decrease is judged to be significantly greater (p<<NUM>) than that seen after placebo (<NUM>%).

After <NUM> weeks of use, it is noted by the experts an amplification of the skin hydrated appearance (<NUM>% significant effect), this value still remains significantly higher (p<<NUM>) to placebo (<NUM>%).

These results confirm those obtained by the quantification by image analysis previously carried out.

As mentioned above, depilation is a triggering or aggravating factor of certain skin disorders including keratosis pilaris. The selected volunteers reported feeling systematically discomfort after wax depiltion of their legs. The effect of the active agent according to the invention was evaluated via a measurement of the TEWL, known to characterize the integrity of the cutaneous barrier. The measurement of the TEWL was carried out in the first (T0) and the second (T4weeks) appointments each time in two stages: before and a little after the standardized withdrawal of a few layers of the stratum corneum using adhesive discs (Dsquames®), this stress mimic reproducibly what hair removal does.

After rupture of the homeostasis of the stratum corneum by this means, it is expected at four weeks of application of the cream according to the invention a lower increase of the TEWL than that observed at T0 and a smaller increase of the TEWL compared to the placebo cream. This indicates a strengthening of the stratum corneum and a lower sensitivity to stress.

The increase in TEWL observed after removal of the skin barrier by adhesives, is reduced by <NUM>% (p<<NUM> vs. T0 and vs. placebo, Table <NUM>) after <NUM> weeks of use of the cream according to the invention. The latter promotes a better resistance of the cutaneous barrier vis-à-vis a stress miming the epilation, this being explained thanks to the observations of the in vitro tests and on explants. At the same time, the application of the placebo cream only resulted in insignificant and more moderate enhancement of <NUM>% (nsd vs. T0).

The small skin disorders change the physiology of the skin which frequently results in the modification of the softness. Less smooth skin often has small dead skin called scales. At time T0 and T4week, a negative imprint of the skin of the legs was performed using Silflo™ silicone polymer. An acquisition was then made by the fringe projection technique (Dermatop, Eotech) on these imprints and an analysis was conducted with a specialized software.

Two representative parameters of the smoothing of the skin were analyzed:.

The analysis of the results (Table <NUM>) shows that the application of the cream according to the invention for <NUM> weeks causes a significant reduction in the depth of the folds of the microrelief and their number. Thus, the overall roughness and the% of surface occupied by the deep network are improved by respectively <NUM>% (p<<NUM>) and <NUM>% (p<<NUM>). These effects are statistically significant compared to the application of a placebo, which does not involve any significant modification of the two previous parameters (<NUM>% and <NUM>%).

Negative imprints of areas with keratosis lesions on the <NUM> volunteers that had some (<NUM> on the upper thighs and <NUM> on the upper arm) were performed. These imprints were then processed as before.

The results show the attenuation of visible lesions of keratosis pilaris through the use of the cream according to the invention. The latter causes a decrease in the number of visible lesions by <NUM>% (p =<NUM> vs. T0). It is also noticed with the cream according to the invention a decrease of <NUM>% of the total perimeter of the lesions which reflects their shrinkage. The comparison with placebo, which does not cause any improvement, shows that the difference is significant for these two parameters (p<<NUM> and p<<NUM>).

This study was carried out on a panel of <NUM> women with a mean age of <NUM> years (<NUM> - <NUM> years old), declaring that their skin was dry and observing a feeling of discomfort and redness during wax depilation of their legs. A period without shaving or depilation was required in order to be able to perform a wax depilation during the evaluation. This study was conducted in open on the legs.

On the day of the evaluation, the volunteers proceeded with a wax depilation of the two half-legs, and immediately after, one leg received a single application of the cream according to the invention while the other leg served as a witness. The perceived effect was evaluated in the short term (between <NUM> and <NUM> minutes after application) and then in the longer term (<NUM> hours after application), using a self-evaluation questionnaire.

For each statement, the volunteers had to choose:.

Furthermore, volunteers were asked if they were satisfied with the product (score from <NUM> to <NUM>, only scores of <NUM> to <NUM> result in satisfaction). Statistical studies were performed using the Khi2 test comparing the% of favorable responses to the% of unfavorable responses.

Claim 1:
Cosmetic or dermatological active ingredient comprising a bacterial extract and a physiologically acceptable medium, characterized in that said extract is an intracellular extract of Salinicoccus hispanicus comprising:
- between <NUM>% and <NUM>% of proteins;
- between <NUM>% and <NUM>% of mineral salts;
- between <NUM>% and <NUM>% of nucleotides;
- <NUM>% to <NUM>% of lipids; and
- between <NUM> and <NUM>% of sugars.