Cosmetic treatment based on pycnidione

Abstract

The treatment is intended for the skin and its appendages to improve their general condition and / or sensory comfort It can be chosen from a treatment for the signs of chronological or premature aging, a moisturizing treatment, a treatment for dull and / or tired skin, a treatment for sensory discomfort of the skin, a treatment for oily and / or acne-prone skin, and / or a hair treatment. Pycnidione can be provided in the form of an extract of the biomass of an in-vitro culture of a microorganism capable of producing pycnidione as a secondary metabolism

Description

[0001] DESCRIPTION

[0002] TITLE: COSMETIC TREATMENT BASED ON PYCNIDIONE

[0003] TECHNICAL FIELD

[0004] The present invention relates to a non-therapeutic cosmetic treatment of the skin and its appendages that can be implemented topically.

[0005] The cosmetic, dermatological and hygiene and personal care product industries are aimed by the invention. They are directed to people and animals, and manufacture and implement active ingredients and final formulas. These industries are constantly looking for new treatments to offer for improving an unsightly state of the skin and / or its appendages or for improving their sensory comfort.

[0006] BACKGROUND ART

[0007] Typically a cosmetic treatment involves preventing or treating unsightly imperfections or discomfort due to chronological or premature aging of the skin and / or its appendages, such as surface homogeneity defects (wrinkles, fine lines, roughness, etc.), colour homogeneity defects (spots, redness, etc.) and radiance of the complexion (loss of radiance, luminosity), flaccidity (appearance of jowls, drooping eyelids, etc.), or sensory discomfort (tingling, tightness, etc.).

[0008] It can also include the treatment of skins or appendages that are not necessarily mature, for example in the case of a preventive treatment, or treatments aimed at all skin types such as moisturizing treatments, improving the radiance of the complexion, or sensory discomfort that can be due to factors other than aging, such as pollution or poor lifestyle.

[0009] In the context of a cosmetic treatment, it will in anyway be a question of offering a curative treatment for diseased skin.

[0010] SUMMARY OF THE INVENTION

[0011] The aim of the present invention is to propose a new cosmetic treatment for the skin (including the scalp and lips) and / or its appendages (including in particular the hair, body hair, eyelashes and eyebrows, nails), advantageously this treatment being adapted to implement one or more active compounds obtained by a biotechnological route within a logic of "natural" products and sustainable development.

[0012] To this end, the present invention proposes the use of pycnidione for non-therapeutic cosmetic treatment of the skin (including the scalp and lips) and its appendages to improve their general condition and / or sensory comfort.

[0013] Pycnidione has been described as having anti-plasmodial activity, anti-cancer activity and more recently antibacterial activity, particularly anti-dandruff via an action on Malassezia-type bacteria (WO2010 / 061185).

[0014] Test results are given below in the description showing the advantageous use of pycnidione for a non-therapeutic cosmetic treatment of the skin and / or its appendages, in particular a treatment of the signs of chronological or premature aging, a moisturizing treatment, a treatment of dull and / or tired skin (including dark circles and bags under the eyes), a treatment of sensory discomforts of the skin, a treatment of oily and / or acne-prone skin (treating a pro-acne condition of the skin) and / or a treatment of brittle and / or split hair.

[0015] More specifically and advantageously, the treatment according to the invention is adapted for:

[0016] - maintaining or improving the mechanical and viscoelastic properties of the skin, including the scalp;

[0017] - strengthening the dermal-epidermal junction (DEJ);

[0018] - combating the harmful effects of glycation (anti-glycation treatment);

[0019] - combating the harmful effects of free radicals (antioxidant or anti-free radical treatment, photoprotection);

[0020] - preventing or reducing skin roughness, fine lines, and wrinkles;

[0021] - preventing or reducing skin sagging (jowls, drooping eyelids); - preventing or reducing pigmentation defects; and / or

[0022] - stimulating the production of hyaluronic acid.

[0023] Other cosmetic activities or benefits may also be considered by those skilled in the art.

[0024] Pycnidione has a bis-tropolone-like chemical structure, corresponding to the following formula (I) in its natural form (stereochemistry described in the scientific publication "Total synthesis and computer investigations of sesquiterpene tropolones reduce stereochemical inconsistencies and resolve an ambiguous biosynthetic relationship”, C. Bemis et al. J. Am. Chem. Soc. 2021, 143, 6006-6017):

[0025] According to the invention, the term "pycnidione" also includes dehydroxypycnidione (in which the hydroxyl group of the 10-carbon ring is replaced by a hydrogen) and eupenifeldin (stereoisomer of pycnidione). Pycnidione is preferred.

[0026] According to the invention, the term "pycnidione" includes pycnidione in a suitable, physiologically acceptable salt form.

[0027] Pycnidione can be produced by chemical synthesis, or it can be derived from a natural source and obtained by in- vitro biofermentation.

[0028] The following examples of microorganisms (fungi) can be used to produce pycnidione as a secondary metabolite by in-vitro biofermentation culture: Neosetophoma cerealis (synonym of Coniothyrium cereale, Coniothyrium cerealis, Neosetophoma samarorum, RKDO834 strain), Neosetophoma sp. (MSX50044 strain), Theissenia rogersii (9203120, 92031201, OS-F69284 (ATCC 74390) strains), Phoma sp. (XZ068 strain (CGMCC 10481), Eupenicillium brefeldianum (ATCC 74184 strain), Kionochaeta pughii (BCC 3878 strain), Kionochaeta ramifera (BCC 7585 strain), Gloeotinia sp. (FKI-3416 strain), Aspergillus hancockii, Entodesmium sp. or Phomopsis tersa (FS441 strain).

[0029] Such strains are available from collections or banks, for example for Neosetophoma cerealis from the Central Bureau for Fungal Biodiversity ("Centralbureau voor Schimmelcultures”, CBS) under number CBS 963.68.

[0030] Preferably according to the invention, the pycnidione is provided in the form of an extract of the biomass of an in- vitro culture of a microorganism capable of producing pycnidione as a secondary metabolite. This extract can be purified to increase the pycnidione content. Preferably, the extract is produced by a strain of Neosetophoma cerealis. According to a particular example, the extract produced in-vitro contains as secondary metabolites a set of tropolones, of which at least 80%, preferably at least 90% by weight, correspond to pycnidione, the remainder consisting of epolone A and / or epolone B which are tropolones such as pycnidione.

[0031] When the in-vitro production route by biofermentation is chosen, pycnidione is produced in the biomass under suitable and optimal culture conditions for the microorganism.

[0032] Cultivation may be carried out on any suitable synthetic medium or natural medium provided that it contains appropriate carbon sources, nitrogen sources, and inorganic salts. If necessary, the medium may be appropriately supplemented with vitamins and other nutrients. Examples of general carbon sources include (but are not limited to) sugars such as glucose, maltose, fructose, sucrose, and starch, alcohols such as glycerol and mannitol, amino acids such as glycine, alanine, and asparagine, and oils and fats such as soybean oil and olive oil. Examples of nitrogen sources include organic nitrogen-containing compounds such as soybean powder, corn liquor, beef extract, peptone, yeast extract, amino acid mixtures, and fish powder, as well as inorganic nitrogen compounds such as ammonium salts and nitrates. Micronutrients in the form of inorganic salts may also be used, for example, calcium carbonate, sodium chloride, potassium chloride, magnesium sulfate, copper sulfate, manganese chloride, zinc sulfate, cobalt chloride, and various phosphates.

[0033] The microorganism can be cultivated at an appropriate culture temperature within a range that allows for its growth and efficient production of pycnidione. The preferred culture temperature is 10°C to 32°C, and more preferably 20°C to 25°C. The pH at the start of the culture is preferably about 6 to 8, and the culture duration is between one day and several weeks.

[0034] Cultivation can be stopped when a sufficient amount of pycnidione is produced, preferably the maximum possible amount.

[0035] Cultivation can be carried out in the solid or liquid phase with appropriate stirring in a bioreactor.

[0036] Appropriate treatment of the biomass (once separated from the supernatant) then allows the recovery of an extract containing pycnidione. Any separation method usually performed in in-vitro culture to extract a metabolite from the biomass can be used to recover this extract, such as extraction with a suitable solvent in which the metabolite can be solubilized.

[0037] More preferably according to the invention, to extract the pycnidione from the biomass, extraction is performed using a solvent and, more preferably, a physiologically acceptable solvent, in particular a glycolated solvent, for example pentylene glycol or butylene glycol, or a mixture thereof. A clarification step to remove any remaining cellular debris is furthermore to be carried out, if necessary, at the end of the treatment.

[0038] This extract comprising pycnidione in an appropriate quantity can be advantageously used as such for the implementation of the invention. The extract obtained can also be purified to increase the pycnidione content, until, if desired, purified pycnidione is obtained at a high purity level, for example having a pycnidione content greater than 80%. The purification of pycnidione can be carried out by chromatography or any other suitable means known to those skilled in the art.

[0039] If the biomass extraction solvent is not a physiologically acceptable solvent, and consequently if the extract cannot be used as such, the latter is freed from all traces of non-physiologically acceptable solvent by an appropriate treatment, and the residue of the extract obtained is then included in a physiologically acceptable medium so that it can be used at the appropriate dosage of pycnidione in the cosmetic treatment according to the invention.

[0040] According to the invention, "Physiologically acceptable" means that the compositions are suitable for topical or transdermal use, in contact with the mucous membranes, nails, scalp, hair, and skin of mammals, and more particularly humans, and that the compositions can be ingested or injected into the skin without risk of toxicity, incompatibility, instability, allergic response, or the like. This "physiologically acceptable medium" forms what is conventionally referred to as the excipient of the composition.

[0041] Preferably, according to the invention, the physiologically acceptable medium may be an alcoholic, glycolic, aqueous, hydroglycolic, or hydroalcoholic medium, or formed by a water-in-oil emulsion, an oil-in-water emulsion, or a microemulsion. Preferably, the physiologically acceptable medium is glycolic, according to the invention.

[0042] The "effective" amount of pycnidione for use in the treatment according to the invention depends on the intended use of the composition containing it, in particular face, hands, body, hair / scalp and it also depends on various factors, such as age, the condition of the person or the extent of the disorder. An effective amount means a nontoxic amount sufficient to obtain the desired effect. As a guide, in a composition intended for a consumer for a treatment according to the invention, the pycnidione, to be present in an effective amount, may range in the composition used relative to the total weight thereof from 0.000001% (0.001 ppm) to 10% (100,000 ppm), preferably from 0.000001% (0.01 ppm) to 1% (10,000 ppm), more preferably from 0.00001% (0.1 ppm) to 0.1% (1000 ppm), more preferably from 0.0001% (1 ppm) to 0.1% (100 ppm), even more preferably from 0.001% (10 ppm) to 0.05% (50 ppm), depending on the intended use of the composition and the desired effect, whether more or less pronounced, and the recommended number of applications.

[0043] All percentages and ratios used in this application are by weight of the total composition and all measurements are made at 25°C, unless otherwise specified.

[0044] Also, for guidance, for a cosmetic facial treatment, the European Cosmetics Directive has set a standard application rate of 2.72 mg / cm2 / day / person for a cream-type composition and 0.5 mg / cm2 / day / person for a body lotion-type composition.

[0045] According to other features, the cosmetic treatment according to the invention may be combined with one or more other treatments targeting the skin or hair, such as light therapy, heat therapy, or aromatherapy.

[0046] According to the invention, it is possible to propose multi-compartment devices or kits intended for implementing the method described above, and which could comprise, by way of example, and without limitation, a composition containing pycnidione in a first compartment, and an additional active ingredient acting jointly in a second compartment, the compositions contained in said first and second compartments being considered herein as a combination composition for simultaneous, separate, or spread over time use, particularly in one of the treatments defined above.

[0047] According to other advantageous features, pycnidione may be used according to the invention in combination with one or more additional active ingredients at effective concentrations advantageously providing a synergistic effect or for activity boosting.

[0048] The additional active ingredients may be chosen, for example, from agents for skin lightening, anti-redness, antispots, slimming, soothing active ingredients for the treatment of sensitive, reactive skin, agents filtering radiation, in particular UVA, UVB, IR, visible light including blue light, humectants, agents for moisturizing, exfoliating, smoothing, toning, anti-aging, anti-wrinkle and fine lines, improving mechanical and elastic properties, radiance of the complexion, detoxifying active ingredients, anti-hairgrowth or on the contrary promoting growth, anti-dandruff, acting on the skin barrier, anti-acne, acting on sebum secretion, mattifying, unifying, anti-inflammatory, antioxidant, anti-free radical, anti-glycation, for the treatment of eye contour (anti-dark circles and for treating undereye bags), promoting blood circulation, peptides, vitamins, ceramides, hyaluronic acid, etc. These active ingredients may be obtained by synthetic route or from plant materials, such as plant extracts or products of plant cell culture, fermentation, bacterial culture or enzyme culture.

[0049] The International Cosmetic Ingredient Dictionary & Handbook published by the Cosmetic, Toiletry, and Fragrance Association, Inc. (CTFA), Washington, D.C., describes a wide variety, without limitation, of cosmetic and pharmaceutical ingredients commonly used in the skincare industry, which are suitable for use as additional ingredients in compositions according to the present invention, as long as they are physically and chemically compatible with the other ingredients of the composition and especially with the active agents of the present invention. Moreover, the nature of these additional ingredients must not impair the benefits of the active agents of the invention. These additional ingredients may be synthetic or natural, such as plant extracts, or be derived from a biotechnological method.

[0050] Other skin, scalp or hair care active agents that are particularly useful in combination with the composition according to the invention can be found in the sales literature of Sederma, Crodarom, Alban Muller and Croda, on the website www.croda.com and www.crodabeauty.com. Mention may also be made, by way of example, of the following commercial active agents: betaine, glycerol, Actimoist Bio 2™ (Active Organics), AquaCacteen™ (Mibelle AG Cosmetics), Aquaphyline™ (Silab), AquaregulK™ (Solabia), Carciline™ (Greentech), Codiavelane™ (Biotech Marine), Dermaflux™ (Arch Chemicals, Inc), Hydra'Flow™ (Sochibo), Hydromoist L™ (Symrise), RenovHyal™ (Soliance), Seamoss™ (Biotech Marine), Argireline™ (trade name of acetyl hexapeptide-3 from Lipotec), spilanthol or an extract of Acmella oleracea known as Gatuline Expression™ (Gattefosse), an extract of Boswellia serrata known as Boswellin™, Deepaline PVB™ (SEPPIC), Syn-AKE™ (Pentapharm), Ameliox™, Bioxilift™ (Silab), PhytoCellTec™ Argan (Mibelle), Papilactyl D™ (Silab), Preventhelia™ (Lipotec), or one or more of the following active ingredients sold by Sederma: Subliskin™, Venuceane™, Moist 24™, Vegesome Moist 24™, Essenskin™, Juvinity™, Revidrat™, Resistem™, Chronodyn™, Kombuchka™, Chromocare™, Calmosensine™, Glycokin factor S™, Biobustyl™, Idealift™, Ceramide 2™, Ceramide A2™, Ceramide HO3™, Legance™, Intenslim™, Prodizia™, Beautifeye™, Pacifeel™, Zingerslim™, Meiritage™, Sebuless™, Apiscalp™, Rubistem™, Citystem™, Neonyca™, NG Shea Butter Unsaponifiables™, Majestem™, Hydronesis™, Poretect™, Crystalide™, Amberstem™, Synchrolife™, Sylverfree™, Feminage™, Ameyezing™, BB-Biont™, Revitalide™, Mel[o]stem™, Luceane™ or mixtures thereof.

[0051] Among the plant extracts (in the form of conventional extracts or prepared by an in vitro method) that may be used as additional active agents, mention may also be made, in particular, of extracts of ivy, for example climbing ivy (Hedera helix), Bupleurum chinensis, Bupleurum falcatum, arnica {Arnica montana L), rosemary {Rosmarinus officinalis N.), calendula {Calendula officinalis), sage {Salvia officinalis L), ginseng {Panax ginseng), Ginkgo biloba, St. John's wort {Hyperycum perforatum), butcher's broom {Ruscus aculeatus L), meadowsweet {Filipendula ulmaria L), orthosiphon {Orthosiphon stamincus Benth.), artichoke {Cynara scolymus), seaweed {Fucus vesiculosus), birch {Betula alba), green tea, kola nut {Cola nitida), horse chestnut, bamboo, Centella asiatica, heather, wrack, willow, pilosella, escin extracts, cangzhu extracts, Chrysanthellum indicum extracts, plants of the Armeniacea genus, Atractylodis platicodon, Sinnomenum, pharbitidis, Flemingia, coleus such as C. forskohlii, C. blumei, C. esguirolii, C. scutellaroides, C. xanthantus and C. barbatus, such as Coleus barbatus root extract, extracts of horehound, Guioa, Davallia, Terminalia, Barringtonia, Trema, Antirobia, Cecropia, Argania, Dioscoreae such as Dioscorea opposita or Dioscorea mexicana, extracts of Ammi visnaga, Siegesbeckia, in particular Siegesbeckia orientalis, plant extracts from the Ericaceae family, in particular extracts of bilberry {Vaccinium angustifolium), Arctostaphylos uva ursi, Aloe vera, sterol-containing plants (particularly phytosterols), Manjistha (extract of plants of the Rubia genus, in particular Rubia cordifolia), Guggal (extract of plants of the genus Commiphora, in particular Commiphora mukul), an extract of kola, chamomile, red clover, Piper methysticum (Kava Kava from Sederma), Bacopa monieri (Bacocalmine™, Sederma) and sea whip, Glycyrrhiza glabra, mulberry, Melaleuca (tea tree), Larrea divaricata, Rabdosia rubescens, Euglena gracilis, Fibraurea recisa hirudinea, Chaparral sorghum, sunflower, Enantia chlorantha, Mitracarpe of the Spermacocea genus, Buchu barosma, Lawsonia inermis L, Adiantium capillus-veneris L, Chelidonium majus, Luffa cylindrica, Japanese mandarin {Citrus reticulata blancovar. unshiu), Camelia sinensis, Imperata cylindrica, Glaucium flavum, Cupressus sempervirens, Polygonatum multiflorum, lovely hemsleya, Sambucus nigra, Phaseolus lunatus, Centaurium, Macrocystis pyrifera, Turnera diffusa, Anemarrhena asphodeloides, Portulaca pilosa, Humulus lupulus, Coffea arabica, Ilex paraguariensis, Globularia cordifolia, Oxydendron arboreum, Albizzia julibrissin, Zingiber zerumbet smith, Astragalus membranaceus, Atractylodes macrocephalae, Plantago lanceolata, Leontopodium alpinum (or eldelweiss), Mirabilis jalapa, Apium graveolens, Marrubium vulgare, Buddleja davidii Franch, Monarda didyma, de Lavandula angustifolia or orchids.

[0052] The compositions of the present invention may include peptides, including, without limitation, di-, tri-, tetra-, penta-and hexapeptides and their derivatives. According to a particular embodiment, the concentration of the additional peptide(s), in the composition, ranges from 1x107% and 20%, preferably from 1x106% and 10%, preferably between 1x105% and 5% by weight. In the context of the present invention, the term "peptide” refers here to peptides containing ten 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 (bio)synthetic peptides. It also refers to compositions that contain peptides and which are found in nature, and / or are commercially available.

[0053] Non-limiting examples of dipeptides for use in the context of the present invention comprises to Carnosine (p- AH), YR, VW, NF, DF, KT, KC, CK, KP, KK, TT, PA, PM or PP.

[0054] Non-limiting examples of tripeptides are RKR, HGG, GHK, GGH, GHG, GKH, KPK, KFK, KavaK, KpAK, KabuK, KacaK, KPK, KMOK, KMO2K (MO2 being a di-oxygenated sulfoxide methionine), PPL, PPR, SPR, QPA, LPA or SPA.

[0055] Non-limiting examples of tetrapeptides are KTFK (SEQ ID NO: 1), GQPR (SEQ ID NO: 2), RSRK (SEQ ID NO: 3), KTAK (SEQ ID NO: 4), KAYK (SEQ ID NO: 5), KFYK (SEQ ID NO: 6), TKPR (SEQ ID NO: 7), AVPG (SEQ ID NO: 8), VPGA (SEQ ID NO: 9), LKLE (SEQ ID NO: 10), ELED (SEQ ID NO: 11) or LLAN (SEQ ID NO: 12).

[0056] Non-limiting examples of pentapeptide are KTTKS (SEQ ID NO: 13) and KTSKS (SEQ ID NO: 14).

[0057] Non-limiting examples of hexapeptides are GKTTKS (SEQ ID NO: 15) or VGVAPG (SEQ ID NO: 16).

[0058] Other peptides for use in the context of the present invention may be selected, this list being not limitative, from: lipophilic derivatives of peptides, preferably palmitoyl (Pal) derivatives or myristoyl (Myr), and metal complexes as aforementioned (e.g., copper complex of the tripeptide HGG or GHK).

[0059] Preferred dipeptides include for example N-Palmitoyl-p-Ala-His, Pal-KT, Pal-RT (Sederma). Preferred tripeptide derivatives include for example the copper derivative of HGG (Lamin™, Sigma), Pal-GHK, Lipospondin (N- Elaidoy l-KFK) and its analogs of conservative substitution, N-Acetyl-RKR-NH2 (Peptide CK+), Pal-KavaK, Pal- KpAlaK, Pal-KabuK, Pal-KacaK, Pal-KMO2K, N-Biot-GHK (Sederma) and derivatives thereof.

[0060] Mention may also be made here of the anti-aging tripeptides of general formula X-Pro*-Pro*-Xaa-Y described in WO2015181688 Xaa selected from Leu, Arg, Lys, Ala, Ser, and Asp, at the N-terminus , X chosen from H, -CO- R1and -SO2-R1and at the C-terminal end Y chosen from OH, OR1, NH2, NHR1 or NR1R2, R1 and R2 being, independently of one another, chosen from a alkyl, aryl, aralkyl, alkylaryl, alkoxy and aryloxy group, which may be linear, branched, cyclic, polycyclic, unsaturated, hydroxylated, carbonylated, phosphorylated and / or sulfurized, said group possibly possessing in its backbone a heteroatom particularly 0, S and / or or N, and Pro* corresponding to Proline, an analogue or derivative thereof; comprising, for example, Myr-PPL-OH and Myr-PPR- OH.

[0061] Here can further be cited also the propigmenting and / or pro-MEC dipeptides and tripeptides of general Formula X-(Xaai)n-Pro*-Xaa2-Y disclosed in WQ2014 / 080376, with n=0, 1 or 2, Xaai an hydrophobic aminoacid selected from Ala, Vai, Met, Leu, Iso, Phe, Pro, and analogues and derivatives thereof; or a polar aminoacid selected from Ser, Thr, Tyr, Asp, Glu and analogues and derivatives thereof; and when n=2 the two aminoacids Xaai being the same or different; Xaa2 being an hydrophobic aminoacid selected from Ala, Vai, Met, Leu, Iso, Phe, and analogues and derivatives thereof, or a basic aminoacid selected from Arg, Lys, His, and analogues and derivatives thereof; at the N terminal end X being selected from H, -CO-R1 and -SO2-R1; at the C terminal end Y being selected from OH, OR1, NH2, NHR1 or NR1R2; R1 and R2 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 0, S and / or N heteroatom in its skeleton and Pro* corresponding to a Proline, analogue or derivative thereof; comprising for example the following peptides Pal-SPR-OH, Pal-PPR-OH, Pal-QPA-OH, Pal-LPA-OH, Myr-SPA-OH, Pal-PM-OH, Pal-PA-OH and Pal-PP-OH. Suitable tetrapeptides derivatives for use as additional peptides according to the present invention include but are not limited to Ela-KTAK (SEQ ID NO: 17), Ela-KAYK (SEQ ID NO: 18), Ela-KFYK (SEQ ID NO: 19), Pal-GQPR (SEQ ID NO: 20) or Pal-KTFK (SEQ ID NO: 21).

[0062] Suitable pentapeptides derivatives for use as additional peptides herein include, but are not limited to, Pal-KTTKS (SEQ ID NO: 22), Pal-KTSKS (SEQ ID NO: 23), Pal-YGGFXaa (SEQ ID NO: 24) with Xaa being Leu or Pro.

[0063] Suitable hexapeptides derivatives for use as additional peptides herein include, but are not limited to, Pal- HLDlIXaa (SEQ ID NO: 25) with Xaa being Trp, Phe, Tyr, Tic, 7-hydroxy-Tic or Tpi, or mixtures thereof, Pal- GKTTKS (SEQ ID NO: 26), Pal-VGVAPG (SEQ ID NO: 27).

[0064] The preferred compositions available commercially and sold by Sederma:

[0065] - tripeptides or a derivative include Biopeptide-CL™, Maxi Lip™, or Procapil™ containing GHK;

[0066] - tetrapeptides or a derivative include RIGIN™, Eyeliss™ containing Pal-GQPR (SEQ ID NO: 20) and an excipient,

[0067] - pentapeptide or a derivative as Matrixyl™ containing Pal-KTTKS (SEQ ID NO: 22).

[0068] It can also be mentioned:

[0069] - the mixture of Pal-GHK and Pal-GQPR (SEQ ID NO: 20) (Matrixyl™ 3000), and

[0070] - the mixture of Pal-GHK and Pal-VGVAPG (SEQ ID NO: 27) (Biobustyl ™)

[0071] The following marketed peptides can be mentioned as well as additional active ingredients:

[0072] - Vialox™, Syn-ake™ (p -Ala-Pro-Dab-NH-Bzl) or Syn-Coll™ (Pal-Lys-Val-Lys-OH) marketed by Pentapharm;

[0073] - Argireline™ (Ac-Glu-Glu-Met-Gln-Arg-Arg-NH2 (INCI name = Acetyl hexapeptide-3) (SEQ ID NO: 28), Leuphasyl™ (Tyr-D-Ala-Gly-Phe-Leu) (SEQ ID NO: 29), Aldenine™ (Gly-His-Lys), Trylagen™ (INCI name = Pseudoalteromonas Ferment Extract, Hydrolyzed Wheat Protein, Hydrolyzed Soy Protein, Tripeptide-10 Citrulline (reaction product of Citrulline and Tripeptide-10 (synthetic peptide constituted of aspartic acid, isoleucine and lysine)), Tripeptide-1), Eyeseryl™ (Ac-beta-Ala-His-Ser-His)(SEQ ID NO: 30), Serilesine™ (Ser-lle-Lys-Val-Ala- Val) (SEQ ID NO: 31) or Decorinyl™ (INCI name: Tripeptide-10 Citrulline = reaction product of Citrulline and Tripeptide-10 (synthetic peptide constituted of aspartic acid, isoleucine and lysine) marketed by Lipotec

[0074] - Collaxyl™ (Gly-Pro-GIn-Gly-Pro-GIn (SEQ ID NO: 32)) or Quintescine™ (Cys-Gly) marketed by Vincience;

[0075] - Cytokinol ™ LS (casein hydrolysate) marketed by Les Laboratoires Serobiologiques / Cognis;

[0076] - Kollaren™ (Gly-His-Lys), IP2000™ (Pal-Val-Tyr-Val) or Meliprene™ (INCI name = Monofluoroheptapeptide-1 : reaction product of acetic acid and a synthetic peptide comprising arginine, glycine, glutamic acid, histidine, norleucine, p-fluorophenylalanine and tryptophan) marketed by I'lnstitut Europeen de Biologie Cellulaire;

[0077] - Neutrazen™ (Pal-His-D-Phe-Arg-NH2) marketed by Innovations; or

[0078] - BONT-L-Peptide™, Timp-Peptide™ or ECM Moduline™ marketed by Infinitec Activos.

[0079] It is also possible to envisage combining the invention with one or more cyclic peptides, in particular those extracted from linseed oil described in the Applicant's patent application WC2019 / 149450.

[0080] More specifically, pycnidione may be combined with at least one of the compounds selected from vitamin compounds, groups B, C, E, F, D, and A, in particular, compounds such as niacinamide or tocopherol, retinoid compounds such as retinol, hyaluronic acid, hexamidine, o-lipoic acid, resveratrol, or DHEA, peptides, ceramides, and chemical or physical UV or IR filters, which are conventional active ingredients used in topical cosmetic compositions.

[0081] The present invention also provides the use of pycnidione for the manufacture of a composition for a cosmetic treatment, as described above.

[0082] A composition according to the invention may be applied to the face, body, neckline, scalp, hair, eyelashes, body hair, in any form or vehicle known to those skilled in the art, in particular in the form of a solution, dispersion, emulsion, paste or powder, individually or as a premix or vehicle individually or as a premix in a bound form, incorporated or adsorbed in vectors such as macro-, micro-, or nanocapsules, macro-, micro- or , nanospheres, liposomes, oleosomes or chylomicrons, macro-, micro-, or nanoparticles or macro-, micro or nanosponges, micro- or nanoemulsions, or adsorbed on organic polymer powders, talcs, bentonites, spores or exines, and other inorganic or organic supports.

[0083] In cosmetics, applications may be proposed particularly in the ranges of skin care for the face, body, hair and body hair and ranges of make-up treatments, in particular eyelashes and eyebrows.

[0084] For example, the galenic form of the composition may be a lotion, a cream, a butter, a milk, a solid form, a foam, a gel, a deodorant, an antiperspirant, a shampoo, a conditioner, a hair mask, a face mask, a shower gel, etc.

[0085] The galenical formulations may enter in different product ranges for personal care and / or beauty products including skin care, cleaning, makeup, cleansing, sunscreen, artificial tanning, pre-shave, shaving or aftershave, moisturizer, humectant, emollient, conditioning, exfoliating, astringent, depilatories or antiperspirant, deodorant, etc.

[0086] The composition may be incorporated onto a non-woven or woven material, with natural or synthetic fibres, wool, or any material intended to come into contact with skin and that may be used in clothing, including tights and socks, shorty, day or night underwear, tissues, handkerchiefs or fabric to exert its cosmetic effect via the contact skin / textile and enable continuous topical delivery (cosmetic-textiles).

[0087] In the particular case of a cosmetic hair treatment according to the invention, the composition may comprise at least one surfactant which may be chosen from anionic, non-ionic, amphoteric and / or cationic surfactants.

[0088] Suitable anionic surfactants include alkyl sulfates, alkyl ether sulfates, alpha-olefin sulfonates, sulfosuccinates, isethionates, acylamides, acylglutamates, alkyl ether carboxylates and alkyl phosphates. The alkyl group preferably comprises between 6 and 30, more preferably between 8 and 20, in particular between 10 and 14, and especially 12 carbon atoms. Alkyl ether sulfates and / or alkyl sulfates are preferred, in particular alkali metals, e.g. their sodium and / or ammonium salts. Lauryl ether sulfate and / or lauryl sulfate are particularly preferred anionic surfactants.

[0089] Suitable nonionic surfactants include fatty alcohol acid or amide ethoxylates, alkanolamides and alkoxylated alkanolamides, monoglyceride ethoxylates, sorbitan ester ethoxylates, alkyl polyglycosides, ethylene glycol monoesters, ethylene glycol diesters, and mixtures thereof.

[0090] Suitable amphoteric surfactants include alkylimino dipropionates, alkylamphoglycinates, alkylamphoproprionates, alkylamphoacetates (mono- and di-), N-alkyl beta-aminoproprionic acids, alkylpolyaminocarboxylates, phosphorylated imidazolines, and mixtures thereof.

[0091] Suitable cationic surfactants include alkyl quaternaries, benzyl quaternaries, quaternary esters, ethoxylated quaternaries, alkylamines, and mixtures thereof. The alkyl group preferably comprises between 6 and 30, more preferably between 8 and 22, and particularly between 10 and 20 carbon atoms.

[0092] The cationic surfactant may also be a polyquaternium (or polyquat) material. Polyquats include acrylamide- and / or dimethylallylammonium chloride-based polymers such as Polyquaternium 6, Polyquaternium 7, etc. Polymeric quaternium ammonium salts of guar gum, such as guar hydroxypropyltrimonium chloride, may be used. Polymeric quaternium ammonium salts of cellulose, such as Polyquaternium 10 and the like, and polymeric quaternium ammonium salts of starch, may also be used.

[0093] The hair care composition may also contain at least one secondary surfactant. If present, the secondary surfactant may be selected from a nonionic, amphoteric, betaine, and / or cationic surfactant.

[0094] Suitable betaines include alkyl betaines, alkylamido betaines, alkyl sultaines, alkylamidosultaines and mixtures thereof. Alkylamido betaines are preferred. The alkyl group preferably comprises between 6 and 30, more preferably between 8 and 20, and in particular between 10 and 14 carbon atoms. The hair composition may also include lubricating agents such as talc, magnesium stearate, and mineral oil, wetting agents, emulsifying and suspending agents, preservatives such as methyl- and propylhydroxybenzoates, sweetening agents, and flavoring agents.

[0095] It may include one or more other standard ingredients or carriers commonly used in hair care products, including shine enhancers, moisturizers, herbal additives, hair strengtheners, vitamin additives, colorants, hair thickeners, fixing and styling agents, ultraviolet absorbers, silicone oils, essential oils and perfumes, thickening or viscosity improving agents, detergents, stabilizers, emollients, chelating or sequestering agents, preservatives, disinfectants, antioxidants / free radical scavengers, antistatic agents, conditioning agents, detangling ingredients, emulsifying or dispersing agents, stimulants, softeners, solvents, carrier, and others.

[0096] It may include additional hair-care active ingredients, either as a support or complement to its activity, including the following active ingredients marketed by Sederma:

[0097] Active ingredient acting on the scalp:

[0098] - Apiscalp™: Apium graveolens (celery) seed extract, titrated in senkyunolide and obtained by supercritical CO2 extraction. Its function is to alleviate dandruff and relieve scalp itching. It moisturizes and reduces sebum production.

[0099] - Hairspa™: A combination of lactitol and xylitol in glycerin to soothe and moisturize the scalp. It helps combat scalp discomfort: dryness, itching, dandruff, and irritation by rebalancing the skin's microflora.

[0100] Active ingredient affecting the quality or color of hair:

[0101] - FruitBio™: A complex of alpha-hydroxy acids obtained by lactic fermentation with the addition of malic and citric acids, combined with green tea extract. This active ingredient smooths the hair cuticles, and the green tea softens them, for smoother and softer hair.

[0102] - Heliogenol™: A hydroglycolic extract of sunflower meal titrated with polyphenols. It has strong anti-free radical activity. It protects and repairs natural and colour-treated hair against the aggression of shampoos and UV radiations.

[0103] - Ceramide A2™: Chemical analogue of ceramide 2, a natural molecule in hair. Its function is to restructure, smooth, and strengthen the hair structure, restoring volume, shine, and vigor to damaged hair.

[0104] - Sylverfree™: Hydroglycerin solution of palmitoyl prolyl proline (Pal-PP). It helps combat hair graying.

[0105] Active ingredient that promotes hair growth:

[0106] - Capigen™: Composed of homotaurine, a synthetic analog of taurine, a natural amino acid, a bacterial filtrate rich in peptides, and a marine-derived sulfomucopolysaccharide extract. It is a hair tonic stimulant and helps prevent hair loss.

[0107] - Procapil™: Combination of a vitamin-enriched matrikine (biotinyl-GHK) with apigenin (a natural flavonoid) and naturally occurring oleanolic acid. It strengthens and rejuvenates the hair follicle to slow hair loss. Hair anchoring is promoted by strengthening the metabolism and follicle structure. Oleanolic acid inhibits 5o-reductase, apigenin improves microcirculation, and biotinyl-GHK stimulates cellular metabolism.

[0108] - Kelisoft™: Chelidonin, a highly purified plant molecule in an excipient. Its function is to reduce hair growth.

[0109] - Hair care ceramides of the SPHINGO HAIR™ range, in particular Sphingo Hair™ Drypure™, a bio-fermentation derived ceramide NP which replenishes the skin with essential longer chains ceramide and promotes healthy scalp and hair barrier structures.

[0110] The hair composition may also comprise an anti-dandruff active agent selected from one of the following: nystatin, cuprimyxin, tolnaftate, candicidin, haloprogin, iodochlorohydroxyquine, clotrimazole, undecylenic acid, propionic acid, caprylic acid, benzoic acid, salicylic acid, griseofulvin, amphotericin B, ketoconazole, miconazole, filipin, hamycin, natamycin, rimocidin, bifonazole, butoconazole, econazole, fenticonazole, isoconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, albaconazole, fluconazole, isavuconazole, itraconazole, posaconazole, ravuconazole, terconazole, voriconazole, abafungin, amorolfine, butenafine, naftifine, terbinafine, anidulafungin, caspofungin, micafungin, iclopirox olamine, flucytosine, crystal violet, piroctone olamine, zinc pyrithione, selenium sulfide, tar and tea tree oil, preferably selected from ketoconazole, zinc pyrithione (ZPT), piroctone olamine, octopirox, salicylic acid, selenium sulfide, coal tar, azelaic acid, grimpazole, salicylic acid, undecylenic acid, and mixtures thereof.

[0111] The present invention thus covers a method of cosmetic, non-therapeutic topical treatment for improving the appearance and general condition of the skin and its appendages, as well as its sensory comfort, comprising the topical application to the skin of a subject in need thereof of an effective amount of pycnidione, the treatment being as defined above and in the examples given below.

[0112] DETAILED DESCRIPTION

[0113] The present invention will be better understood in light of the following detailed description of exemplary embodiments.

[0114] 1- Example of preparation of an extract containing pycnidione

[0115] Microorganism used: Neosetophoma cerealis strain available from the Central Bureau for Fungal Biodiversity (CBS).

[0116] A pre-inoculum step is performed using cryopreserved spores or mycelium inoculated into a specific pre-inoculum culture medium (medium containing a gelling agent, sugar, and nitrogen in the form of peptone or yeast extract). Several pre-inoculum steps are performed to increase the biomass.

[0117] When the pre-inoculum has grown sufficiently, the biomass produced is used to inoculate the bioreactor with a culture medium based on rice and micronutrients. In the bioreactor, also called a producer, the biomass is amplified with the concomitant production of pycnidione.

[0118] After 7 days of culture in the producer, pycnidione production is maximal.

[0119] The biomass is then harvested using solid / liquid separation techniques such as centrifugation, decantation by centrifugation, or filtration through a filter press. The supernatant is discarded.

[0120] The harvested biomass is mixed with a solvent intended to form a physiologically acceptable excipient, consisting of a mixture (20 / 80 w / w) of pentylene glycol (PTG) and butylene glycol (BG), with stirring for 1 hour at 60°C to extract the pycnidione into this excipient. After extraction, the cellular debris are removed by clarification using a known solid / liquid separation technique, as mentioned above.

[0121] Analysis of the dry extract determines a final concentration of total tropolones in the extract of 1200 ppm, including approximately 1100 ppm of pycnidione, the target active molecule according to the invention, and the remaining tropolones including, in particular, Epolone A and Epolone B.

[0122] 2- Example of preparation of an active ingredient for the treatment according to the invention

[0123] The PTG / BG extract prepared in point 1 below, comprising a physiologically acceptable excipient, constitutes an active ingredient that can be used in a cosmetic formula at a defined weight percentage depending on the desired effect, whether more or less pronounced, as explained above in the description. Various cosmetic formulas using this cosmetic ingredient are described further in the description in point 4.

[0124] 3- In-vitro efficacy tests

[0125] The tests were performed with either the dry extract of the biomass prepared in point 1 above diluted in DMSO, or pure pycnidione (approximately 82%) also diluted in DMSO. The ppm values indicated in the tables below correspond to the ppm values of the pycnidione molecule.

[0126] All statistical analyses were performed using a Student's t-test to determine the significance of the results of pairwise comparisons (control versus treated). 3.1- Stimulation of hyaluronic acid synthesis

[0127] Hyaluronic acid is an essential component of the skin. It is present in the dermis and epidermis, and thanks to its unusual three-dimensional structure, it is capable of capturing a thousand times its weight in water and thus occupying a very large volume relative to its molecular weight. The intracellular spaces of the extracellular matrix are filled with water-swollen hyaluronic acid, which gives the skin good viscoelastic properties.

[0128] Thus, by stimulating hyaluronic acid synthesis, hydration is maintained or increased, resulting in more resilient and elastic skin, with a smoother texture and a more comfortable feel, without tingling or tightness.

[0129] Protocol:

[0130] Human keratinocytes (HK) are cultured to subconfluence and then placed in contact with the active ingredient according to the invention. Following this contact, hyaluronic acid synthesis is assessed using an ELISA assay. Estimating cell counts using the Hoechst method allows to homogenise the results.

[0131] Results

[0132] Variation in hyaluronic acid production by keratinocytes compared to the control.

[0133] [Table 1]

[0134] The results show that pycnidione significantly increases hyaluronic acid synthesis in keratinocytes.

[0135] 3.2- Strengthening the dermo-epidermal junction (DEJ)

[0136] The DEJ ensures cohesion between the epidermis and dermis. During skin aging, whether normal or accelerated by external factors, there is a decrease in the synthesis of DEJ components, particularly laminins and collagen VII, leading to disorganization with repercussions on the skin's elasticity and resilience, and a loss of its vitality.

[0137] Protocol

[0138] KH cells are cultured to sub-confluence and then placed in contact with the active ingredient according to the invention (or not, for control cases). Following this contact, collagen VII and laminin synthesis are assessed using ELISA kits. Estimating cell counts using the Hoechst method normalizes the results.

[0139] Results

[0140] Variation in laminin and collagen VII production by keratinocytes compared to the control:

[0141] [Table 2] The results show that pycnidione significantly increases the production of laminins and collagen VII in keratinocytes.

[0142] 3.3- Anti-glycation:

[0143] The human body needs sugar to produce energy. However, the sugar consumed is never fully utilized by the body for this beneficial purpose. A residual portion of this absorbed sugar will react non-enzymatically with the amino groups of proteins, nucleic acids, or lipids to create advanced glycation end products (AGEs), called glycotoxins, or AGEs, which accumulate over time in tissues. Glycated molecules have their functional, enzymatic, and structural properties altered, which impact the proper functioning of cells and the body.

[0144] In the skin, AGEs are found not only in the dermis but also in the epidermis, all the way to the stratum corneum. Tissues are particularly affected when their proteins, such as collagen, elastin, fibronectin, and laminins, are long- lived. This results in altering the mechanical and elastic properties of the extracellular matrix (ECM) of the dermis, which becomes less flexible, more rigid, but also more flaccid and less reactive.

[0145] Furthermore, within skin cells, mitochondrial proteins are also glycated, causing decreased efficiency in the production of adenosine triphosphate (ATP) and impaired energy production.

[0146] AGEs therefore represent a major source of cellular dysfunction, and their accumulation tends to alter the properties and performance of tissues and cells: reduced flexibility, reduced mobility, reduced responsiveness, and reduced energy production. Glycation leaves its mark on the face, for example, with dark circles under the eyes, puffy bags, a dull complexion and drawn features, and less responsive, tired, and flaccid skin. A study has also shown that the quantity of AGEs and hair quality are linked. A reduction in protein content and an increase in AGEs result in poorer hair quality with reduced resistance to breakage.

[0147] Glycation therefore visually translates on the skin by the appearance of wrinkles and fine lines, dull, flaccid, lackluster, tired-looking skin, skin lacking tone and suppleness, and on the hair as poorer quality hair, split ends and brittleness.

[0148] Protocol

[0149] The study of non-enzymatic glycation is carried out between a model protein, serum albumin, which serves as a target, and an edible reducing sugar derived from fruit. The protein is progressively glycated (bound to the sugar) irreversibly, in the presence or absence of the product according to the invention. This modification is monitored by fluorescence.

[0150] Results

[0151] Variation in AGE production compared to the control:

[0152] [Table 3]

[0153] The results show the strong anti-glycating potential of pycnidione.

[0154] 3.4- Decreased synthesis of pro-inflammatory molecules:

[0155] The skin is subjected to constant stress (exposure to UV rays, smoke, pollutants, etc.), some of which trigger a direct or indirect inflammatory response. The uncontrolled or constant inflammatory response, although low- intensity, leads to the production of cytokines designed to attract or stimulate other cells, triggering a cascade of reactions. The resulting pro-inflammatory microenvironment alters skin homeostasis and gradually leads to the modification or even destruction of cell and tissue biomolecules. It also disrupts the integrity of the skin barrier. Thus, inflammatory mediators IL-6 and IL-8 are known to induce premature aging through micro-inflammation. Furthermore, sensitive and irritated skin is characterized by an abnormally high secretion of these cytokines. By reducing the production of these cytokines following stress, the invention demonstrates a preventive antiinflammatory effect. The cells are better protected. From a cosmetic point of view, the treatment according to the invention makes it possible to prevent discomfort, such as tightness, and unsightliness, such as micro-redness of the skin, for example due to external aggressions such as pollution, radiation or cold.

[0156] Protocol

[0157] Normal human keratinocytes (NHK) were seeded and placed in contact or not with the active ingredient to be tested at different concentrations for 48 hours and irradiated with UVB. Non-irradiated plates were treated in parallel. The amounts of IL-6 and IL-8 synthesized were measured in the culture supernatants using ELISA assays. Cell viability was estimated at the end of the culture using Hoechst staining on fixed cell lawns (cell number estimation).

[0158] Results

[0159] Variation in IL-6 and IL-8 synthesis after keratinocyte stress compared to the control:

[0160] [Table 4]

[0161] The results show that pycnidione induces a decrease in IL-6 and IL-8 synthesis after stress (UVB conditions).

[0162] 3.5- Decrease in intracellular production of reactive oxygen species (ROS):

[0163] It is now recognized that it is essentially the accumulation of oxygenated free radicals that is at the origin of the biochemical disorders themselves responsible for aging. Whether generated by UV rays (via a complex cascade of reactions), by the various pollutants that interact with the skin chronically or by natural aging, these free radicals will cause skin inflammation punctually and over a short period of time, and throughout the body and over a longer period of time skin aging. Under so-called normal conditions, on young, healthy skin that is relatively protected from external aggressions, there is a balance between the production of oxygenated free radicals in the body and the antioxidant defenses responsible for neutralizing them. These defense systems are made up of either enzymes (SOD, catalane, glutathione peroxidase), or classic antioxidants (vitamin C, vitamin E, glutathione, carotene, etc.). When an imbalance occurs between production and neutralization, there is an overabundance of oxygenated free radicals causing oxidative stress. This will result in damage to DNA, proteins and cellular lipids, which will undergo oxidation, causing alterations in their structure and function. This results in the appearance of signs of premature aging, damage to the dermis and epidermis with loss of their mechanical and viscoelastic properties, thinning of the skin, sagging of the skin, fine lines, wrinkles, spots, etc.

[0164] Protocol

[0165] Normal human fibroblasts (NHF) are seeded and placed in contact with the product for 24 hours. After removal of the active ingredient, the cells are rinsed and loaded with the DCFH-DA probe, which becomes fluorescent after metabolism by cellular enzymes in the presence of ROS. This fluorescence is further enhanced when a stressor (H2O2) is applied to the cells. An active ingredient will reduce both basal and induced fluorescence (ROS production).

[0166] Cell viability is assessed in parallel using Hoechst staining (DNA intercalating reagent). Results

[0167] Variation in ROS synthesis by fibroblasts compared to the control, with and without oxidative stress:

[0168] [Table 5]

[0169] Pycnidione significantly reduces the generation of reactive oxygen species (ROS) following H2O2 stress, as well as basally (unstressed conditions), without dose-dependent effects.

[0170] Pycnidione helps combat the appearance of intracellular ROS, known for their role in skin aging.

[0171] 3.6- Anti-Lipoperoxidation:

[0172] Lipoperoxidation, or lipid peroxidation, is the peroxidation of the lipid membranes surrounding cells and mitochondria, leading to their weakening and the generation of oxygen reactive species (free radicals) through autocatalysis, amplifying the damage.

[0173] Protocol

[0174] Lipid peroxidation is assessed using a test that monitors the occurrence of peroxidation (measuring the quantity of conjugated dienes formed) of lipid membranes in the form of liposomes after oxidative UVA radiation. Reduction by an antioxidant is sought. The product is then used in this test after irradiation.

[0175] Results

[0176] Variation in lipoperoxidation compared to the control:

[0177] [Table 6]

[0178] The results show the strong potential of pycnidione to fight against lipoperoxidation and therefore its harmful effects on cell membranes and their dynamism.

[0179] 3.7- Slowing the proliferation of C. acnes:

[0180] The skin microbiota, a highly complex ecosystem composed of a variety of living microorganisms (bacteria, yeasts, viruses, and parasites), has several functions: defence, skin barrier, and immune system regulation. It is important to maintain its balance by preventing certain species from causing damage to the skin by overgrowth. This is the case, for example, with C. acnes (Cutibacterium acnes), the acne bacterium. Although part of the skin's normal microflora, if it multiplies too quickly, it promotes the proliferation and migration of keratinocytes, participates in the formation of free radical species such as superoxide anion, and triggers a cascade of reactions that results in the production of pro-inflammatory molecules, thus contributing to the development of acne.

[0181] By slowing down or inhibiting the growth of C.acnes, the epidermis is protected against acne. This effect is particularly interesting for cosmetic treatment of so-called oily and acne-prone skin. This type of skin often corresponds to oily skin in adolescence due to excess sebum for hormonal reasons. By preventing or limiting the proliferation of the C. acnes bacteria, the occurrence of waste in the hair sheath in the hair follicle is prevented. It is important because it is in the hair sheath that the bacteria develop anaerobically by feeding on this sebum and by producing waste, including dead cells, and then leading to obstruction of the hair follicle sheath (blackheads), inflammation and finally acne pimples, the treatment of which is a matter of dermatology. Reducing the quantity of C. Acnes therefore helps prevent the development of a pro-acne state.

[0182] Protocol

[0183] Suspensions of C. acnes of equivalent density are cultured in a suitable medium in the presence (test case) or absence (control case) of the test product, under anaerobic conditions, at 37°C. After 48 to 72 hours of incubation, the growth of the bacterial population is measured by reading the optical density (OD) at 600 nm.

[0184] Results

[0185] Variation in C. acnes proliferation compared to the control:

[0186] [Table 8]

[0187] The results show that pycnidione induces a significant slowdown in the proliferation of C. acnes bacteria responsible for a pro-acne state.

[0188] 4- Examples of cosmetic formulas

[0189] Various cosmetic formulas are described below. Additional active ingredients, where appropriate supporting and / or complementing 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 may be of any category depending on their function, the place of application (body, face, neck, bust, hands, hair, eyelashes, eyebrows, body hair, etc.), the desired final effect and the targeted consumer, for example an antioxidant, moisturizing, nourishing, protective, smoothing, remodelling, volumizing agent, an agent acting on the radiance of the complexion, anti-spot, anti-dark circles, anti-glycation, slimming, soothing, muscle relaxant, anti-redness, antistretch mark agent, etc. They are mentioned above in the description.

[0190] Ingredient according to the invention: ingredient prepared in paragraph 2) above (extract prepared in-vitro containing approximately 1100 ppm of pycnidione in a mixture of butylene glycol and pentylene glycol).

[0191] Recommended effective amount of ingredient: for example, between 1% and 15%, preferably between 1% and 10%, more preferably between approximately 1% and 5%, which corresponds to between approximately 10 ppm and 60 ppm of pycnidione, indicated at 3% (approximately 35 ppm) in the examples of formulas below. 4.1- Light cream formula, adapted for the face

[0192] [Table 9]

[0193] Preparation:

[0194] Weigh part A. Weigh and mix part B. Slowly add part B to part A while stirring and mix well for 30 min. Heat part A+B to 75°C in a bain-marie. Weigh and heat part C to 75°C in a bain-marie. Add part C to part A+B while stirring vigorously. Add part D to part A+B+C until about 45°C. Add part E to the previous part while stirring. Where applicable, add part F to the previous part, mix well.

[0195] A light, non-sticky cream is obtained that can be used, for example, as a facial cream for a relaxing massage application.

[0196] 4.2- Cream formula adapted for a day cream

[0197] [Table 10]

[0198] Preparation:

[0199] Weigh part A, mix well with normal stirring. Weigh and mix part B. Pour part B into part A with normal stirring for one hour. Heat part A+B to 75°C in a bain-marie. Weigh and heat part C to 75°C in a bain-marie. Add part C to part A+B, stirring quickly. Weigh and add part D to the previous part at about 45°C, mix well. Weigh and add part E to the previous part, mix well. If necessary, weigh and add part F to the previous part, mix well.

[0200] A white opaque cream with a light and soft touch is obtained that can be used for day care. The active ingredient Crystalide® can be used to support the activity to help repair the skin and give a radiant complexion.

[0201] 4.3- Cream formula, suitable for oily, acne-prone skin

[0202] [Table 11]

[0203] Preparation:

[0204] Weigh part A and mix. Weigh and mix part B. Add part B to part A while stirring for 1 hour. Heat part A+B to 75°C in a bain-marie. Weigh and heat part C to 75°C in a bain-marie. Add part C to part A+B while stirring vigorously. Weigh and mix part D. Add part D to the previous part, mix well. Add part E, mix well. Where applicable, add part F, mix well.

[0205] An opaque, off-white viscous cream is obtained, suitable for oily skin thanks to its dry feel and rapid absorption. With the combination with the ingredient Fruitbio™ a mild chemical exfoliant treatment is added to furthermore smooth the texture of the skin.

[0206] 4.4- Cream formula, suitable for hands

[0207] [Table 12]

[0208] Preparation:

[0209] Weigh Part A and mix under normal stirring. Weigh and mix Part B. Pour Part B into Part A under normal stirring for one hour. Activate Part A+B by stirring vigorously for 1 min. Add Part C into Part A+B, stir well. Heat Part A+B+C to 75°C in a bain-marie. Weigh and heat Part D to 75°C in a bain-marie. Weigh and add Part E to Part D well before creating the emulsion. Add Part D+E to the previous part, mix well under rapid stirring. Weigh and mix Part F. Add Part F to the previous part at about 45°C, mix well. Weigh and add Part G, to the previous part, mix well. Weigh and add Part H, to the previous part, mix well.

[0210] A cream is obtained which is perfectly suited to the treatment of dry hands thanks to the ingredient according to the invention and the additional association with a ceramide, a natural lipid constituent of the skin.

[0211] 4.5- Cream formula, suitable for face and neck

[0212] [Table 13] Preparation:

[0213] Weigh part A. Weigh and mix part B. Slowly add part B to part A while stirring and mix well for 30 min. Heat part A+B to 75°C in a bain-marie. Weigh and heat part C to 75°C in a bain-marie. Add part C to part A+B while stirring vigorously. Add part D to part A+B+C while stirring. If necessary, add part E to the previous part while stirring. Mix well during the cooling phase.

[0214] A pale-yellow cream is obtained, ideal for the face and neck, which is quickly absorbed by the skin.

[0215] 4.6- Cream formula, suitable for a day cream for the neckline

[0216] [Table 14]

[0217] Preparation:

[0218] Weigh part A and mix under normal stirring. Weigh and mix part B. Pour part B into part A under normal stirring for one hour. Activate part A+B by stirring vigorously for 1 min. Weigh part C. Add part C to part A+B, stir well. Heat part A+B+C to 75°C in a bain-marie. Weigh and heat part D to 75°C in a bain-marie. Weigh and add part E to part D well before creating the emulsion. Add part D+E to the previous part, mix well under rapid stirring. Weigh and add part F to the previous part, mix well. Weigh and add part G to the previous part, mix well.

[0219] An opaque cream emulsion with a soft and light texture is obtained, quickly absorbed by the skin, suitable for a large area such as the decollete. 4.7- Cream lotion formula

[0220] [Table 15]

[0221] Preparation:

[0222] Weigh part A, mix well under normal agitation. Weigh and mix part B. Add part C to part B, stir well. Weigh and add part D to B+C, mix well. Weigh part E, mix well. Adjust pH of B+C+D to pH + / -5.40 with part E, mix well. Slowly add part B+C+D+E into part A, stirring rapidly.

[0223] A white opaque fluid emulsion is obtained, forming a super concentrated lotion comparable to an W / O emulsion with a rich cream feel.

[0224] 4.8- Hydrogel formula

[0225] [Table 16]

[0226] Preparation:

[0227] Heat part A to 80°C in a bain-marie with stirring for 1 hour. Weigh part B and mix well. Slowly add part B to part A with stirring at 80°C in a bain-marie. Allow part A+B to heat to 80°C for 1 hour in a bain-marie with stirring. Add part C to part A+B in a bain-marie at 80°C. Add part D to part A+B+C and mix well. Add part E where applicable and mix well. Pour immediately into the molds, at a warm temperature.

[0228] A hydrogel is obtained capable of providing an instant refreshing and soothing sensation while the active ingredient(s) act in depth to soothe and protect the skin in the long term. The addition of Crodateric CAB 30™ as a mild surfactant allows the skin to be gently cleansed with water at the end of use.

[0229] 4.9- Concentrated serum formula

[0230] [Table 17] Preparation:

[0231] Weigh part A, mix well with normal stirring. Weigh and mix part B. Pour part B into part A with normal stirring for one hour. Weigh and add part C into part A+B, with rapid stirring. Weigh part D, mix well. Add part D to the previous part, stir well. Weigh and add part E to the previous part, mix well. If applicable, weigh and add part F to the previous part, mix well.

[0232] An emulsion is obtained forming a super concentrated serum that resembles a gel and has the feel of a rich cream. In combination, the ingredient according to the invention and the Synchrolife™ ingredient provide a complete regenerating night care.

[0233] 4.10- Paste formula

[0234] [Table 18]

[0235] Preparation:

[0236] Weigh part A and heat to about 65°C. Weigh part B. Add it to part A and mix well while stirring. Weigh part C and add it to part A+B while stirring. Weigh part C and add it to the previous part at 50°C. Weigh and add part D. Mix well. If applicable, weigh and add part E, mix well. Pour immediately into the pot.

[0237] An opaque beige paste is obtained that can be used as a concealer and as a screen against urban pollution. 4.11- Cream formula, suitable for day care around the eyes

[0238] [Table 19]

[0239] Preparation:

[0240] Weigh and mix part A while stirring. Weigh part B. Add part B to part A while stirring normally. If applicable, weigh part C. Add part C to part A+B and mix well. Weigh part D. Weigh part D and mix well while stirring normally. Add part A+B+(C) to part D while stirring quickly and mixing well.

[0241] An opaque, off-white viscous cream is obtained, perfectly suited to eye contour care, firming and illuminating this area. The Feminage™ active ingredient reinforces the treatment by acting on sagging skin.

[0242] 4.12- Spray formula

[0243] [Table 20]

[0244] Preparation:

[0245] Weigh part A. Weigh and mix part B. Pour part B into part A under normal stirring. Weigh and add part C into part A+B, stir well. Weigh and add part D to the previous part, mix well. Weigh and mix part E. Add part E to the previous part, mix well. If applicable, weigh and add part F to the previous part, mix well.

[0246] A clear, pale-yellow liquid is obtained that can be used as a spray with a light texture. The addition of the BB- Biont™ ingredient also helps regulate the microbiome and smooth out acne marks. The formula is particularly suitable for application to treat skin worn a mask.

[0247] 4.13- Lotion formula, adapted for the body

[0248] [Table 21]

[0249] Preparation:

[0250] Weigh part A, mix well. Heat part A to 75°C in a bain-marie. Weigh part B, mix well. Add part B to part A while stirring rapidly for one hour at 75°C in a bain-marie. Mix part A+B while stirring vigorously for 1 minute. Weigh and heat part C to 75°C in a bain-marie. Add part C to part A+B while stirring vigorously. Weigh part D, mix well. Adjust the pH =5.80+ / -0.10 with Part D, at approximately 45°C, mix well. Add part E to the previous part, mix well. If applicable, add part F to the previous part, mix well.

[0251] An opaque fluid cream with a fresh and silky texture is obtained. It can be used as an after-hair removal lotion, for example in combination with the anti-hair regrowth active ingredient Kelisoft™.

[0252] 4.14- Solid stick formula

[0253] [Table 22]

[0254] Preparation:

[0255] Weigh and heat part A to 75°C in a bain-marie. Weigh part B and add to part A. Weigh and mix part C. Heat part C to 75°C in a bain-marie. Add part C to part A+B while stirring quickly. Weigh part D and mix. Add part D to the previous part at about 50°C while stirring quickly. Pour to form the stick in a mold immediately, at about 45°C which becomes solid at room temperature.

[0256] A solid, opaque, light-yellow stick is obtained. It can be used for a quick and practical touch of care. The combination of the ingredient according to the invention with the PoreTect™ ingredient makes it possible to couple a moisturizing, anti-aging and specific care action for distended pores. In day care, the Solaveil™ CT-300 ingredient also protects against damage that can be caused by UV rays.

[0257] 4.15- Rich cream formula with a whipped texture

[0258] [Table 23]

[0259] Preparation:

[0260] Weigh part A, mix well. Weigh part B, mix well. Add part B to part A, stirring rapidly for one hour. Heat part A+B to 75°C in a bain-marie. Weigh and heat part C to 75°C in a bain-marie. Add part C to part A+B, stirring vigorously. Weigh part D, mix well. Add part D to the previous part, at about 45°C, stir well. Add part E to the previous part, stir well. Add part F, if applicable, to the previous part, stir well.

[0261] After 24 hours of rest, stir the emulsion with a U-shaped paddle for one hour.

[0262] A pale yellow opaque viscous cream with a whipped mousse texture is obtained. The cream is rich but leaves the skin with a comfortable non-greasy sensation. 4.16- Roll-on Lip formula

[0263] [Table 24]

[0264] Preparation:

[0265] Weigh part A and heat to about 75 °C. Add part B and mix well for 30 minutes while stirring. Weigh part C and heat to about 75 °C. Add part C to part A+B while stirring at high speed. Weigh part D and mix well. Adjust the pH to 6.00-6.20 with part D when the temperature is below 45 °C, mix well. Weigh and add part E to the previous part, mix well. Weigh part F and add it to the previous part, mix well.

[0266] A dark red fluid emulsion is obtained, compatible with a roll-on type of application that offers a restorative, moisturizing and protective cosmetic treatment thanks to its active ingredients, and an aesthetic visual effect of volume and shine to the lips. 4.17- Shampoo formula

[0267] [Table 25]

[0268] Preparation:

[0269] Weigh part A. Mix well. Add the ingredients of part B, one after the other, to part A+B while stirring. Mix well.

[0270] Weigh part C, mix well. Add part C to part A+B with normal stirring. Add part D to the previous part with normal stirring. Add part E to the previous part while stirring normally.

[0271] A clear pale-yellow fluid gel is obtained, forming a shampoo to gently cleanse and treat the hair and scalp.

[0272] 4.18- Leave-on spray formula for hair application

[0273] [Table 26] Preparation:

[0274] Weigh part A and mix. Weigh part B and mix well. Weigh part C and mix well. Mix part B into part C. Pour part B+C into part A with normal helical agitation. Add part D into the previous part with normal helical agitation.

[0275] An opaque off-white fluid emulsion with a texture suitable for application by spraying is obtained to be deposited lightly on the hair and scalp.

[0276] 4.19- Mask formula for hair application

[0277] [Table 27] Preparation:

[0278] Weigh and heat part A to 85°C in a bain-marie. Weigh and heat part B to 90°C in a bain-marie. Weigh part C and mix well. Pour part C into part A with normal stirring. Disperse part B into part A + C with vigorous stirring, mix well. If necessary, pour part D, at approximately 40°C, into the previous part while stirring. If necessary, add part E, mix well. If necessary, add part F to the previous part, mix well. Add part G, mix well. Add part H, mix well.

[0279] A yellow opaque viscous emulsion is obtained forming a thick cream suitable for a silicone-free hair and scalp mask that can include a range of active ingredients such as the active ingredient according to the invention for a complete cosmetic treatment.

[0280] 4.20- Serum formula for hair application

[0281] [Table 28]

[0282] Preparation:

[0283] Sprinkle the carbomer in the water and let it swell for 30 minutes. Weigh part B. Add part B to part A while stirring. Weigh and mix part C. Neutralize with part C to part A + B. Homogenize well with stirring. Weigh and mix part D. Pour part D into part A + B + C while stirring. Add part E and mix well. Add part F and mix well.

[0284] A colorless fluid gel forming a serum with a light, non-greasy texture is obtained.

Claims

CLAIMS1 . Use of pycnidione for a non-therapeutic cosmetic treatment of the skin and its appendages to improve their general condition and / or sensory comfort.

2. Use according to claim 1 , characterized in that the pycnidione is provided in the form of an extract of the biomass of an in-vitro culture of a microorganism capable of producing pycnidione as a secondary metabolite, said extract being optionally purified to increase the pycnidione content.

3. Use according to claim 2, characterized in that the extract is derived from the biomass of an in-vitro culture of a strain of Coniothyrium cerealis.

4. Use according to any one of the preceding claims, characterized in that the pycnidione is added to a physiologically acceptable medium.

5. Use according to claim 4, characterized in that the physiologically acceptable medium corresponds to the biomass extraction solvent.

6. Use according to any one of the preceding claims, characterized in that the treatment is topical.

7. Use according to any one of the preceding claims, characterized in that the treatment is chosen from:- a treatment for the signs of chronological or premature aging,- a moisturizing treatment,- a treatment for dull and / or tired skin,- a treatment for sensory discomfort of the skin,- a treatment for oily and / or acne-prone skin, and / or- a hair treatment.

8. Use according to any one of the preceding claims, characterized in that the treatment is suitable for maintaining or improving the mechanical and viscoelastic properties of the skin, including the scalp.

9. Use according to any one of the preceding claims, characterized in that the treatment is suitable for preventing or reducing roughness, fine lines, and wrinkles of the skin.

10. Use according to any one of the preceding claims, characterized in that the treatment is suitable for preventing or reducing skin sagging.11 . Use according to any one of the preceding claims, characterized in that the treatment is suitable for stimulating the production of hyaluronic acid.

12. Use according to any one of the preceding claims, characterized in that the treatment is suitable for strengthening the dermis-epidermis junction (DEJ).

13. Use according to any one of the preceding claims, characterized in that the treatment is suitable for combating the harmful effects of glycation.

14. Use according to any one of the preceding claims, characterized in that the treatment is suitable for combating the harmful effects of free radicals.

15. Use according to any one of the preceding claims, characterized in that the hair treatment is suitable for treating brittle or split ends, and / or preventing and / or treating pigmentation defects.

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