Composition comprising a myrtle extract and a Tripterygium wilfordii extract for combating acne in non-lesional areas of acne-prone skin

Abstract

The present invention relates to a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically acceptable or dermatologically acceptable excipient, for its use in the treatment and / or prevention of acne in non-lesional areas of acne-prone skin.

Description

Title of the invention: Composition comprising a myrtle extract and a Tripterygium wilfordiï extract for combating acne in non-lesional areas of acne-prone skin TECHNICAL FIELD OF THE INVENTION

[0001] The present invention relates to cosmetic or dermatological compositions comprising a myrtle extract and a Tripterygium wilfordii extract, for their use in the field of acne, and more particularly in the treatment and / or prevention of acne in non-lesional areas of acne-prone skin (i.e. outside lesional areas with visible inflammation: papules, pustules and nodules), in particular by acting directly on the phylotypes of C. acnes. STATE OF THE ART

[0002] Acne is a chronic inflammatory skin condition of the pilosebaceous unit leading to the formation of comedones, affecting the face, scapular region, arms and intertriginous regions. It is the main cause of the most common dermatoses. It is important not to trivialize this condition and to treat it correctly because it can have disabling psychosocial consequences, particularly due to the formation of scars.

[0003] Acne is a multifactorial pathology. Indeed, various risk factors can lead to the development of acne or accentuate it. These factors concern hormonal impregnation and genetic heritage but also external environmental factors, such as pollution and UV rays. Finally, lifestyle can also influence the appearance or amplification of acne, namely diet and stress. There are several forms of acne. It can first manifest itself by the appearance of simple comedones (blackheads and whiteheads). This is called retentional acne. But these retentional lesions can also develop into more severe inflammatory lesions: papules, pustules and nodules. In this case, we speak of inflammatory acne. However, it is now established that inflammation is present at all stages of acne, even the earliest, i.e., the subclinical stages: this is called micro-inflammation.Classically, these two forms coexist: this is mixed acne, affecting nearly 60% of acne sufferers.

[0004] Common acne or acne vulgaris, also called juvenile polymorphic acne, is the most common and includes four stages:

[0005] - Stage 1 corresponds to comedonal or retentional acne and is characterized by a large number of open and / or closed comedones and microcysts; - Stage 2, or papulopustular acne, is mild to moderate in severity and is characterized by the presence of open and / or closed comedones, microcysts, but also red papules and pustules. It mainly affects the face and leaves some scars; - Stage 3, or papulo-comedonian acne, is more severe and extends to the back, chest and shoulders. It is accompanied by a large number of scars; - Stage 4, or nodulocystic acne, presents with nodules and also large, painful purple pustules. It is accompanied by numerous scars.

[0006] In its mildest form, acne affects almost every human being. Its frequency is highest at puberty, but it can appear for the first time as early as 7 to 9 years old and up to ages exceeding 40. It is common to suffer from acne even after the age of 25. Acne also affects both men and women. During puberty, under the influence of hormonal secretions and in particular androgens, but also associated with various external factors, an overproduction of sebum called hyperseborrhea is observed. In subjects predisposed to acne, this environment is conducive to the development of the key acne bacterium, Cutibacterium acnes (C. acnes) (formerly called Propionibacterium acnes).This bacterium metabolizes skin triglycerides into irritating fatty acids via lipases that attack the follicle wall and the surrounding dermis, also produces various enzymes and chemoattractants of phagocytic immune cells, and stimulates the production of pro-inflammatory cytokines by different types of cells (sebocytes, keratinocytes, monocytes, in particular) that aggravate inflammation. This bacterium plays a pivotal role in acne, notably by stimulating the local inflammatory response (Dagnelie et al., Journal of the European Academy of Dermatology 2019, 33(12), 2340-2348). It has long been thought that the fight against this bacterial species was a priority in the treatment of acne. The use of various topical antimicrobials is still widely used today (benzoyl peroxide, erythromycin, triclosan). But recently, researchers have realized that the goal is not to eradicate C.acnes which is a commensal bacterium, necessary for tissue homeostasis, but to restore a balance because acne is associated with a loss of diversity of C. acnes phylotypes with a predominance of the pro-pathogenic phylotype IA1. This phylotype is pro-pathogenic and has a strong capacity to organize itself into a biofilm conferring greater virulence to this bacterium. This virulence can be measured by quantifying the virulence factors produced by C. acnes. Contrary to what has long been thought, acne is not associated with a multiplication of C. acnes but with a modification of the ratio of phylotypes and a loss of richness in these phylotypes.

[0007] Similarly, more or less prolonged systemic antibiotic therapy was sometimes associated with the treatment depending on the severity of the condition (tetracyclines, doxycycline). Dermatologists are now turning more towards local anti-inflammatories, sebum regulators.

[0008] Frequent failures of all these treatments have been observed, often due to a high proportion of resistant strains of C. acnes. This resistance may be the consequence of an organization of bacterial populations into biofilms. Biofilms are bacterial cell communities integrated into an extracellular matrix excreted by microorganisms, composed of sugar polymers and called glycocalyx. Sessile (biofilm-associated) bacteria are phenotypically and physiologically different from planktonic (free-living) bacteria. Studies have confirmed the ability of C. acnes to form biofilms both in vitro (Holmberg et al., Clin. Microbiol. Infect. 2009, 15, 787-795) and in vivo on medical devices (Craig et al., J. Am. Acad. Dermatol. 2007, 722-724).

[0009] Thus, there is still a need to provide more effective treatments for acne that do not present adverse effects in the patient. In particular, there are currently no agents that act both on the lesion areas and also on the subclinical areas, i.e. areas where acne is not yet visible. The aim of future treatments is therefore no longer to kill the C. acnes bacteria but to rebalance the skin microbiota, which is essential for skin homeostasis. Summary of the invention

[0010] The object of the present invention is to meet these needs. Indeed, the inventors have demonstrated, quite unexpectedly, that cosmetic or dermatological compositions comprising a myrtle extract and a Tripterygium wilfordii extract had the capacity to reduce the pro-pathogenic IC phylotype, found only in acne-prone skin, and to increase the IB phylotype associated with healthy skin. These compositions are therefore capable of rebalancing the C. acnes phylotypes in non-lesional areas (i.e. excluding papules, pustules and nodules) of acne-prone skin, in particular in subjects with mild to moderate acne.

[0011] The present invention therefore relates to a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient, for its use in the treatment and / or prevention of acne in non-lesional areas of acne-prone skin.

[0012] The present invention also relates to the use of a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically suitable excipient. logically acceptable, for the treatment and / or prevention of acne in non-lesional areas of acne-prone skin.

[0013] The present invention also relates to the use of a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient, for the preparation of a medicament intended for the treatment and / or prevention of acne in non-lesional areas of acne-prone skin.

[0014] The present invention also relates to a method for treating and / or preventing acne in non-lesional areas of acne-prone skin comprising the administration, in particular the topical application to the non-lesional areas, to a person in need thereof of an effective amount of a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient.

[0015] The present invention also relates to a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient, for its use in the prevention of acne.

[0016] The present invention also relates to the use of a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient, for the prevention of acne.

[0017] The present invention also relates to the use of a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient, for the preparation of a medicament intended for the prevention of acne.

[0018] The present invention also relates to a method for preventing acne comprising the administration, in particular the topical application, to a person in need thereof of an effective amount of a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient. DETAILED DESCRIPTION OF THE INVENTION

[0019] The invention relates to a cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, in particular with at least one cosmetically or dermatologically acceptable excipient, for its use in the treatment and / or prevention of acne in non-lesional areas of skin. acne, or in the prevention of acne, acne being induced by C. acnes. Myrtle extract

[0020] Common myrtle, Myrtus communis L., is a shrub of the Myrtaceae family. The myrtle extract according to the present invention is produced more particularly from the leaves of Myrtus communis. Preferably, it is an apolar fraction of aerial parts, and more particularly the leaves, of myrtle.

[0021] By "apolar fraction" is meant a fraction of an apolar extract, typically an apolar extract treated with activated carbon so as to eliminate chlorophylls.

[0022] By "apolar extract" is meant an extract capable of being obtained using an apolar extraction solvent, such as for example ethyl acetate, isopropyl acetate or a mixture thereof.

[0023] By "aerial parts" according to the invention is meant the parts of the plant located above the ground, for example, the leaves, the stems, the petioles and / or the inflorescences, in particular the leaves.

[0024] The extract according to the present invention can be obtained by extraction using a solvent or mixture of solvents (called extraction solvent) chosen from:

[0025] - alcohols such as ethanol, methanol, isopropanol, - ketones including acetone and methylethylacetone, - hexane, - methylene chloride, - isopropyl ether, - ethyl or isopropyl acetate, - and their mixtures;

[0026] or by extraction using supercritical CO2.

[0027] According to one embodiment, the myrtle extract can be obtained by extraction using isopropyl acetate, preferably from the aerial parts, and in particular the leaves, of Myrtus communis.

[0028] Thus, the plant (or any part thereof) is brought into contact with the extraction solvent. Once the extraction is carried out, the plant-extraction solvent mixture is filtered so as to separate the solvent phase from the plant residues (called marc). The marc thus obtained is rinsed, typically with the same solvent as the extraction solvent, and the rinsing solvent thus obtained is mixed with the solvent phase so as to obtain an extraction juice.

[0029] Advantageously, the extraction juices, obtained after filtration and rinsing of the marc, are decolorized by the addition of activated carbon, which allows the elimination of chlorophylls.

[0030] The extract can also be stabilized by adding an antioxidant such as butylhydroxytoluene or alpha tocopherol, in particular in quantities between 0.05 and 1% by weight of dry extract.

[0031] The extract according to the present invention has in particular the characteristic of comprising, and in particular of being rich in, myrtucommulones and ursolic acid. Myrtucommulones A, B', D, B, isos (isosemimyrtucommulmone) and S (semimyrtucommulone) are advantageously present in the extract and are in particular the main myrtucommulones present.

[0032] Advantageously, the content of total myrtucommulones in the extract is between 3 and 10% by weight of dry extract.

[0033] The ursolic acid content is between 10 and 30%, preferably > 15% by weight of dry extract.

[0034] These molecules carry at least in part the activity claimed within the framework of the present invention.

[0035] In a particular embodiment of the invention, the myrtle extract will preferably be as described in patent application EP 1 112 079, the teaching of which is incorporated into the present application, or according to example 1 of the present application. This document EP 1 112 079 describes the antibacterial properties of the Myrtus communis extract and its applications in cosmetic or dermatological compositions.

[0036] The myrtle extract as used in the present invention has also been the subject of patent FR 2 992 862, its anti-biofilm activity against C. acnes being described therein. Extract of Tripterygium wilfordii

[0037] Tripterygium wilfordii is a medicinal plant belonging to the Ce-lastraceae family. Terpenes are among the most active components of the plant and are located mainly in the roots of the plant. These include pentacyclic triterpenes, such as Tingenin A (also called Tingenone or Maytenine), Tingenin B (also called 22beta-hydroxy-tingenone), Celastrol, Pristimerine and Tripterygone. These molecules are described in patent application EP 3 454 875, the teaching of which is incorporated into the present application.

[0038] In one embodiment, the extract of Tripterygium wilfordii may be an extract comprising, in particular enriched in, pentacyclic triterpene(s), such as Tingenin A, Tingenin B and / or Celastrol, and in particular a mixture of Tingenin A, Tingenin B and Celastrol. The extract may optionally comprise components (including active components) derived from the plant other than pentacyclic triterpenes.

[0039] By "pentacyclic triterpene" according to the invention is meant a pentacyclic triterpene naturally produced by the cells of the plant Tripterygium wilfordii and in particular Celastrol of formula Chem. I, Tingenin A (also called Tingenone or Maytenin) of formula Chem. II, Tingenin B (also called 22beta-Hydroxy-tingenone) of formula Chem. III, Pristimerine of formula Chem. IV, and / or Tripterygone of formula Chem. V, preferably Celastrol, Tingenin

[0040] A and / or Tingenin B, in particular Celastrol or a mixture of Tingenin A, Tingenin B and Celastrol. [Chem.I]

[0041]

[0042]

[0043] CH3 [Chem. II] [Chem. III] [Chem. IV] O HSC HV f 1 0¾

[0044] [Chem.V] ^COOH

[0045] By “crude extract” is meant an extract directly obtained from a plant, in this case Tripterygium wilfordii.

[0046] By "enriched extract" of Tripterygium wilfordii is meant an extract of Tripterygium wilfordii in which the quantity of pentacyclic triterpene(s), in particular Tingenin A, Tingenin B and / or Celastrol, is greater than 30% by weight, in particular greater than 50% by weight relative to the quantity of pentacyclic triterpenes in a dry crude extract.

[0047] In one embodiment, the extract, in particular the enriched extract, of Tripterygium wilfordii for use according to the invention comprises between 90% and 100% of pentacyclic triterpene(s) by weight relative to the total weight of the dry extract, in particular the dry enriched extract.

[0048] The extracts, in particular the crude or enriched extracts, can be obtained from any part of the Tripterygium wilfordii plant, in particular the roots, the seeds or the aerial parts.

[0049] Alternatively, the extract of Tripterygium wilfordii can be obtained by plant cell cultures of these plants. In such a case, the extract may in particular be obtained from the supernatant, suspension or biomass of said cell cultures, as described in particular by Coppede et al., Plant Cell Tiss Organ Cuit, 2017, 118,33-43.

[0050] In a preferred embodiment, the extract of Tripterygium wilfordii is an extract comprising, in particular enriched in, pentacyclic triterpene(s), and capable of being obtained according to the following process:

[0051] i. A proliferation phase of Tripterygium wilfordii cells in a proliferation medium, ii. An elicitation phase by adding an elicitation cocktail to the cell culture obtained in step (i), said elicitation cocktail comprising at least one monocarboxylic compound type elicitor and at least one biotic elicitor, and iii. The preparation of an extract comprising, in particular enriched in, pentacyclic triterpene(s) from the cell culture obtained in step (ii).

[0052] By “Tripterygium wilfordii cells” according to the invention, we mean the cells of any part of the plant: seeds, roots, aerial parts, and especially aerial parts, and more particularly leaves.

[0053] By "Tripterygium wilfordii cell proliferation phase" is meant according to the invention a phase in which the Tripterygium wilfordii cells are suspended in a proliferation medium and in conditions adapted to their proliferation. These cells may in particular be obtained from calluses before being suspended. If necessary, the cell suspensions may be regularly re-seeded in order to maintain them in proliferation conditions.

[0054] By "callus" according to the invention, we mean a cluster of dedifferentiated cells, also called stem cells or meristematic cells.

[0055] By "about", it is meant in the present description that the concerned value may be lower or higher by 10%, especially 5%, particularly 1%, than the indicated value.

[0056] Callus induction can be obtained by any method known to those skilled in the art. The calli according to the invention can in particular be obtained in the manner described below. Callus induction from an explant of plant tissue, especially an aerial part such as a leaf, of Tripterygium wilfordii, is well known to those skilled in the art.

[0057] Callus induction can in particular be carried out by:

[0058] - obtaining an explant of plant tissue, for example a piece of leaf about 1 cm2 in size, - culturing the explant on a solidified proliferation medium (for example by adding 4 to 12 g / L of agar, for example about 8 g / L of agar, to the proliferation medium according to the invention) - incubation, especially in the dark, at a temperature of approximately 25-30°C, for example at approximately 27 to 28°C.

[0059] The following steps: cell proliferation phase, elicitation phase and phase of preparation of the extract comprising, in particular enriched in, pentacyclic triterpenes are described in patent EP 3 454 875, the teaching of which relating to the implementation of these steps is incorporated into the present application. Cosmetic or dermatological compositions

[0060] The invention relates to cosmetic or dermatological compositions useful in the treatment and / or prevention of acne in non-lesional areas of acne-prone skin, or in the prevention of acne.

[0061] For the purposes of the present invention, the term "non-lesional areas" means areas of the skin that do not have papules, pustules or nodules. This is also referred to as the subclinical stage of local inflammation.

[0062] The invention preferably relates to cosmetic or dermatological compositions according to the invention being in a form suitable for topical application.

[0063] The cosmetic or dermatological compositions according to the invention can thus be presented in the forms which are usually known for topical administration, that is to say in particular lotions, shampoos, balms, mousses, gels, dispersions, emulsions, sprays, serums, masks, creams or sticks, in particular with excipients allowing in particular, for some, penetration in order to improve the properties and accessibility of the active ingredients.

[0064] The cosmetic or dermatological composition according to the invention will advantageously comprise a myrtle extract which is an apolar fraction comprising myrtu-commulones and ursolic acid. In particular, the myrtucommulones comprise myrtucommulones A, B', D, B, isosemimyrtucommulone and semimyrtu-commulone.

[0065] The cosmetic or dermatological composition according to the invention will advantageously comprise an extract of Tripterygium wilfordii which comprises at least one pentacyclic triterpene as defined above, and in particular chosen from Tingenin A, Tingenin B, Celastrol, Pristimerine, Tripterygone and mixtures thereof, and in particular chosen from Tingenin A, Tingenin B, Celastrol and mixtures thereof, and in particular a mixture of Tingenin A, Tingenin B and Celastrol. The extract of Tripterygium wilfordii will advantageously be an extract of Tripterygium wilfordii comprising, in particular enriched in, pentacyclic triterpene(s), such as Tingenin A, Tingenin B and / or Celastrol, and more particularly an extract of Tripterygium wilfordii comprising, in particular enriched in, pentacyclic triterpene(s) capable of being obtained by the process described above and which comprises the following steps:

[0066] i. A proliferation phase of Tripterygium wilfordii cells in a proliferation medium, ii. An elicitation phase by adding an elicitation cocktail to the cell culture obtained in step (i), said elicitation cocktail comprising at least one monocarboxylic compound type elicitor and at least one biotic elicitor, and iii. The preparation of an extract comprising, in particular enriched in, pentacyclic triterpenes from the cell culture obtained in step (ii).

[0067] The cosmetic or dermatological composition according to the invention will comprise in particular between 0.01 and 1% of an extract of Tripterygium wilfordii by weight of dry extract relative to the weight of the total composition, in particular between 0.05 and 0.8%, in particular between 0.1 and 0.5%, in particular between 0.2 and 0.4% by weight of dry extract relative to the weight of the total composition. According to a preferred embodiment, the cosmetic or dermatological composition according to the invention comprises approximately 0.3% of an extract of Tripterygium wilfordii by weight of dry extract relative to the weight of the total composition.

[0068] The extract of Tripterygium wilfordii will advantageously comprise 90% or more of pentacyclic triterpene(s) by weight relative to the total weight of the dry extract. In an advantageous embodiment, the extract of Tripterygium wilfordii will comprise between 1 and 20% of Tingenin A by weight relative to the total weight of the dry extract, between 1 and 20% of Tingenin B by weight relative to the total weight of the extract and at least 60% of Celastrol by weight relative to the total weight of the dry extract.

[0069] The cosmetic or dermatological composition according to the invention will advantageously comprise between 0.01 and 1% of a myrtle extract by weight of dry extract relative to the weight of the total composition, in particular between 0.05 and 0.8%, in particular between 0.08 and 0.4%, in particular between 0.08 and 0.2% by weight of dry extract relative to the weight of the total composition. According to a preferred embodiment, the cosmetic or dermatological composition according to the invention comprises approximately 0.1% of a myrtle extract by weight of dry extract relative to the weight of the total composition.

[0070] The myrtle extract will advantageously comprise between 3 and 10% of myrtu-commulones by weight relative to the total weight of the dry extract and / or between 10 and 30% of ursolic acid by weight relative to the total weight of the dry extract.

[0071] The following examples illustrate the invention without limiting its scope. DESCRIPTION OF FIGURES

[0072] [Fig.l] represents the HPLC chromatogram obtained for the extract “CCV” of Tripterygium wilfordii according to example 2 containing the following pentacyclic triterpenes: Celastrol, Tingenin A and Tingenin B. EXAMPLES Example 1: Preparation of a myrtle extract

[0073] 1 kg of crushed myrtle leaves are extracted with 5 volumes of isopropyl acetate under reflux stirring for 1 hour. After filtration and rinsing of the marc, the extraction juices are decolorized by adding activated carbon. After filtration, the decolorized filtrate is concentrated to 2 liters then dried over ethanol until the isopropyl acetate is removed. The aqueous phase obtained is then deodorized by heat treatment, then dried by freeze-drying.

[0074] 1 kg of myrtle leaves provides approximately 25 g of dry myrtle extract. This It contains 7% myrtucommulones and 25% ursolic acid.

[0075] Example 2: Obtaining a plant cell culture extract (CCV) from Tripterygium wilfordii enriched with pentacyclic triterpenes

[0076] A culture is carried out in a Wave type reactor (volume 5 L) from Sartorius Stedim Biotech (Germany). The reactor is inoculated with a suspension of cells from Tripterygium wilfordii from an Erlenmeyer flask. The proliferation medium has, for example, the composition indicated below:

[0077] Macroelements: NH4NO3 at 1650 mg / L, KN03 at 2500 mg / L, CaCl2.2H2O at 440 mg / L, MgSO4.7H2O at 370 mg / L, KH2PO4 at 130 mg / L;

[0078] Microelements: Kl at 0.41 mg / L, H3BO3 at 6.2 mg / L, MnSO4.4H2O at 22.3 mg / L, ZnSO4.H2O at 7.5 mg / L, Na2MoO4.2H2O at 0.25 mg / L, CuSO4.5H2O at 0.025 mg / L, CoC12.6H2O at 0.025 mg / L, FeSO4.7H2O at 19.85 mg / L, Na2EDTA.2H2O at 26.64 mg / L;

[0079] Vitamins: myo-inositol 50 mg / L, nicotinic acid 0.25 mg / L, pyridoxine-HCl 0.25 mg / L, thiamine-HCl 0.25 mg / L;

[0080] Carbon source: sucrose at 30 g / L;

[0081] Plant hormones: NAA acid at 0.35 mg / L, 2,4 D acid at 0.575 mg / L, kinetin at 0.083 mg / L.

[0082] The pH of the medium is adjusted to pH 6 ± 0.5 (by adding KOH, IM) before an appropriate sterilization treatment, for example autoclave at 121°C for a minimum duration of 20 minutes or by sterilizing filtration on 0.2 µm.

[0083] With continuous stirring, after reaching the maximum biomass approximately 17 days later, elicitation is carried out. The elicitation cocktail is then added to the Erlenmeyer flask in the proliferation medium using stock solutions made in dimethyl sulfoxide. The composition of the elicitor cocktail makes it possible to obtain the following concentrations in the elicitation medium (+ cells): sodium pyruvate 1.5 g / L, potassium pyrophosphate 0.44 g / L, 2iP 0.0004 g / L, methyl jasmonate 0.036 g / L and chitin 2 g / L. The culture is stopped after 15 days of elicitation. The majority of the biomass is recovered by filtration of the cell suspension with a nylon filter (20-50 pm). From 5 L of suspension, approximately 1925 g of biomass is recovered. This biomass is extracted with ethyl acetate (or isopropyl acetate) in a proportion of 2:1 (Volume: Weight) relative to the weight of biomass (here 3850 mL of solvent for 1925 g of biomass).The biomass / solvent mixture is then subjected to physical extraction, by sonication or grinding. The organic phase is then recovered after maceration with stirring. The addition of the solvent (followed by maceration with stirring and recovery of the organic phase) is repeated twice. The solvent was concentrated under vacuum, then the concentrate was solubilized in pentylene glycol, and the mixture was placed under vacuum to remove the residual organic solvent. A solution called "CCV extract" is then obtained containing the following pentacyclic triterpenes: Celastrol, Tingenin A and Tingenin B, as determined by the HPLC chromatogram shown in [Fig.l].

[0084] HPLC chromatography conditions: Alliance liquid chromatography apparatus (Waters 2695 version 2.03); Sunfire C18 column, 100Â, 5 pm (4.6 mm X 150 mm); water / acetonitrile solvent gradient, flow rate 3 ml / min; triterpene detection at / . 460 nm.

[0085] Example 3: Formulation of a composition useful in the context of the present invention

[0086] Formulation A in cream form has the composition described in Table 1 below.

[0087] [Tables 1] Ingredients Quantity (% by weight) Water 65.47 Glycerin 99.5% 5.00 Pentylene glycol 3.00 Caprylic / Capric Triglycerides 45-55 8.00 Dicaprylyl carbonate 5.00 Benzoic acid 0.20 Cetearyl alcohol / cetearyl mixture 4.00 Polymethyl methacrylate 3.00 Polyacrylate- 13 / Polyisobutene / Polysorbate 20 1.20 Sodium hydroxide 0.03 Stearyl glycyrrhenate 0.10 Glyceryl laurate 0.60 Niacinamide 4.00 Tripterygium wilfordii extract 0.30 Myrtus communis leaf extract 0.10 Example 4: Clinical study

[0088] The pathogenesis of acne involves an interaction of four main factors: overproduction of sebum, hyperproliferation of keratinocytes, modification of the skin microbiota, notably involving the bacterium Cutibacterium acnes (C. acnes), and the induction of local inflammation. C. acnes, the most represented bacterial species in the sebaceous zone of the skin, is thought to play a major role in the pathogenesis of acne (Dréno et al. Am.J. Clin. Dermatol. 2020, 21 (Suppl 1): 18-24). Interestingly, recent studies have demonstrated that acne results from a specific loss of C. acnes phylotype diversity, rather than from hyperproliferation of C. acnes (Pécastaings et al. J. Eur. Acad. Dermatol. Venereol. 2018, 32 (Suppl 2): ​​15-23). ​​Indeed, a predominance of phylotype IA was observed. This phylotype has been described as having more virulent activity in acneic skin than in non-acneic skin, via its greater capacity to form biofilms. A recent study showed that, in inflammatory acne lesions, the proportion of C. acnes phylotype IA increases while phylotypes IB and II decrease (Foumiere et al. Microorganisms 2020, 8(11):1752). Interestingly, phylotypes IB and II, predominantly expressed in healthy skin, do not alter tissue integrity and are not pro-pathogenic in the context of acne (Laclaverie et al Exp. Dermatol. 2021, 30(3): 347-357). Finally, the IC phylotype is specific to acneic skin, no IC phylotype is detectable on normal / healthy skin (McDowell et al PloS One 2013, 13; 8(9) e70897).Based on biofilm formation (directly associated with bacterial virulence), phylotype IA shows the highest biofilm formation, followed directly by phylotypes IC and II, while phylotypes IB and III, phylotypes present only on healthy skin, show the lowest biofilm formation (Kuehnast et al Int. J. Med. Microbiol. 2018, 308(8): 1027-1035).

[0089] The study is divided into two parts:

[0090] - Evaluate the long-term efficacy (56 days) of the application of formulation A according to example 3 in 41 patients with facial acne judged to be mild to moderate; - Estimate the evolution of the microbial flora, in particular of C. acnes, in 29 of these patients (having therefore followed the same protocol). First part of the study:

[0091] 41 patients aged 12 to 35 are recruited, including 22 adolescents (12-17 years). They have mild to moderate acne with a total of 15 to 50 acne lesions, 10 to 30 non-inflammatory lesions (open and closed comedones), and 5 to 20 inflammatory lesions (papules and pustules) on the face, excluding the nasal area. Subjects have at least one post-inflammatory erythema or post-inflammatory hyperpigmentation.

[0092] Two applications per day for 56 days of a formula A (example 3) are carried out.

[0093] A first inclusion visit on D0 allows for an initial assessment. The product application period is at home from D1 to D56. A second visit is made on D8, a window of ± 1 day is authorized for this second assessment. A third visit is made on D29, a window of ± 2 days is authorized for this third assessment. A fourth visit or end-of-study visit is scheduled on D57, a window of ± 2 days is authorized for this last assessment, provided that the patients continue to participate in the study.

[0094] Different criteria are evaluated:

[0095] - Variations in the number of total acne lesions on the face between D0, D8, D29 and D57, the counting being done according to the Lucky method; - The evolution of the number of retentional lesions (open and closed comedones, also called blackheads and microcysts respectively) and inflammatory lesions (papules and pustules) on the face between D0, D8, D29 and D57, the counting being done according to the Lucky method; - Change in acne severity according to the investigator's Global Acne Assessment between D0, D8, D29 and D57; - The overall evolution of acne by the subjects at D8, D29 and D57 on a 6-point scale (-1: worsening; 0: no change; 1: slight improvement; 2: moderate improvement; 3: good improvement; 4: very good improvement); - Clinical evolution (by the investigator) of skin softness, on an 11-point scale (0 to 10) between D0, D8, D29 and D57. Results

[0096] The evolution of acne lesions is summarized in Table 2 below.

[0097] [Tables2] D0 (avg ± wk) D8 (avg + wk) D29 (avg + wk) D57 (avg + wk) Blackheads 7.6+1.1 6.2+1.1 4.1 +0.8*** 4.0 + 0.8*** Microcysts 13.6+1.3 11.8 + 1.3* 10.7 + 1.2* 9.1 + 1.3** LRG 21.1 + 1.1 18.0 + 1.3*** 14.9 + । q*** 13.1 + 1.5*** Papules 5.0 + 0.3 5.1 + 0.6 3.5 + 0.4** 3.1 + 0.5*** Pustules 3.2 + 0.5 2.4 + 0.3 1.7 + 0.3*** 1.7 + 0.3** LIG 8.2 + 0.5 7.4 + 0.6* 5.2 + 0.5*** 4.8 + 0.7*** Total lesions 29.3 ± 1.3 25.4 ± 1.6*** 20.0 ± 1.5*** 17.9 ± 2.0***

[0098] LRG: global retentional lesions; LIG: global inflammatory lesions

[0099] avg ± sem: mean ± standard deviation

[0100] *P<0.05 vs D0; **P<0.01 vs D0; ***P<0.001 vs D0.

[0101] Under these conditions, after 56 days of twice-daily application, formula A exhibits a significant anti-acne effect on the number of retentional and inflammatory lesions (Table 2). This anti-acne effect appears after 8 days of treatment on global lesions. The protective effect appears to progress throughout the treatment.

[0102] This improvement during the study is clearly evidenced by the global acne assessment (EGA), shown in Table 3 below.

[0103] [Tables3] D0 (avg ± wk) D8 (avg ± wk) D29 (avg + wk) D57 (avg + wk) EGA score 2.5 + 0.1 2.4 + 0.1 2.1 +0.1** 1.9 + 0.2***

[0104] **P<0.01 vs D0; ***P<0.001 vs D0

[0105] At the end of the study, 49% of patients showed a visible improvement in acne.

[0106] The assessment of the overall development of acne by the subjects themselves (6-point scale) is presented in Table 4 below, in % of subjects meeting the criterion of development of their acne at D8, D29 and D57.

[0107] [Tables4] EGA D8 D29 D57 -1: worsening 0 3 8 0: no change 38 20 17 1: slight improvement 41 32 26 2: moderate improvement 16 29 31 3: good improvement 5 16 13 4: very good improvement 0 0 5

[0108] After 8 days of application, 62% of subjects felt an overall improvement (stages 1 to 4) and 5% of patients found that their skin had a good improvement. At the end of the study, 75% of subjects felt an overall improvement (stages 1 to 4) and 18% found that their skin had at least a good improvement.

[0109] The results on the estimation of skin softness (scale from 0 to 10, 0: very rough skin; 10: very soft skin) during the study are summarized in Table 5 below.

[0110] [Tables 5] D0 (avg + wk) D8 (avg + wk) D29 (avg + wk) D57 (avg + wk) Softness effect 4.2 + 0.2 4.8 + 0.2*** 5.2 + 0.3*** 6.1 +0.3*** [YES] ***P<0.001 vs. D0

[0112] Formula A therefore has a smoothing effect by improving the clinical score assessed by a dermatologist

[0113] The inventors have clearly demonstrated that formula A exhibits remarkable anti-acne clinical activity. Second part of the study:

[0114] The study is carried out on 29 subjects, 15 adolescents (12-17 years) and 14 adults. Swab samples are taken from the forehead, on non-lesional areas, that is to say that papules and / or pustules are excluded, we speak of subclinical stage of local inflammation. The samples are taken on D1, before the first application of the product, and on the last day of the study (D 57).

[0115] These are the same patients (29 of the 41 patients in the full study), so they have mild to moderate acne and the treatment protocol is therefore the same. Sequencing of the C. acnes phylotype

[0116] After extraction of DNA from the swabs, the analysis of the phylotype of C. acnes is carried out on the GeT Genomics platform (INRAE) using SLST (Singe-Locus Sequence Typing) with Illumina Miseq technology.

[0117] From the extracted DNA, the SLST target gene described by Scholz et al. PLOS 1994, 9(8) el04199, is amplified by PCR using modified oligonucleotides. The sequence of each DNA fragment is then obtained by high-throughput sequencing. Analysis of the sequences obtained and their comparison with the list of reference sequences from the database http: / / medbac.dk / slst / pacnes allows the relative abundances of each C. acnes phylotype to be determined within the different samples. Sequencing is performed on a MiSeq sequencer (Illumina). Sequences showing 100% homology between them are grouped into unique sequences, then into OTUs (operational taxonomic unit: threshold of 100%) which will be identified later. Rare OTUs (present 1 time with 1 reading) are deleted. Bioinformatics analysis of sequencing data allows identification of the phylotype of C. acnes present.Analysis for phylogenetic affiliation down to the phylotype level was performed using bioinformatics tools for processing large amounts of sequence data (Mothur). Three phylotype pairs cannot be discriminated with this method: phylotype DI is indistinguishable from phylotype D2, phylotype E3 is indistinguishable from phylotype E6, and phylotype Fl is indistinguishable from phylotype FIL.

[0118] All the SLST phylotypes obtained are grouped into the following 5 phylotypes:

[0119] - IA: sequences A to F

[0120] - IC: G sequences

[0121] - IB: H sequences

[0122] - II: K Sequences

[0123] - III: L sequences

[0124] The statistical analysis carried out on the sequencing data gives an index of diversity and richness on the phylotypes of C. acnes: IA, IB, IC, II and III. Results

[0125] The distribution of C. acnes phylotypes at the start of the study is shown in the following Table 6.

[0126] [Tableauxô] Phylotypes Abundance (%) AI 83.2 IB 9.0 CI 1.9 II 5.7 III 0.2

[0127] As shown in Table 6, at the start of the study (before the first application), phylotype IA is by far the most represented in the non-lesional areas (i.e. excluding papules and pustules) of the acne-affected skin of the forehead of the volunteers, followed by phylotypes IB, II, IC and III.

[0128] The statistical analysis of the evolution of the phylotypes between D0 and D57 is represented in Table 7 below.

[0129] [Tables7] Evolution of phylotypes between D0 and D57 P value IA 0.69 NS IB 0.03 * IC 0.01 ** II 0.69 NS III 0.78 NS

[0130] *: p<0.05; **p<0.01; NS: not significant

[0131] The application of formulation A does not modify the abundance of phylotypes IA, II and III after 57 days of treatment. On the other hand, as shown in the data in Table 7, the twice-daily application of formulation A for 57 days induces a significant increase in phylotype IB, increasing from an abundance of 8.95 to 9.28%, and a significant decrease in phylotype IC, increasing from an abundance of 1.90 to 1.32%.

[0132] A different abundance of C. acnes phylotypes is observed in patients acne patients compared to healthy controls, with a predominance of the IA phylotype in acne patients (Pécastaings et al. 2018).

[0133] In this study, the inventors show that phylotype IA is the most represented, followed by phylotypes IB, II, IC and III, as has already been described on the inflammatory areas of acne patients. But, they obtain new results on the abundance of C. acnes phylotypes on non-lesional areas of acne skin.

[0134] Indeed, in a completely unexpected and surprising manner, the inventors show that the application of formula A induces a significant increase in the non-pathogenic phylotype IB and significantly reduces the acne-specific phylotype IC. These results therefore suggest that formulation A is effective in an acne context. This is the first time that significant effects on the acne-specific phylotype IC have been demonstrated.

[0135] In this study, the inventors, using a single-locus sequence typing scheme for C. acnes, confirm the link between the presence of certain C. acnes phylotypes and acne, and more particularly in the non-lesional areas of subjects with mild to moderate acne. In addition, the inventors demonstrate that prolonged application of formulation A has a positive impact on the non-lesional areas of acne-prone skin by acting directly on the C. acnes phylotypes, by decreasing the pro-pathogenic IC phylotype and increasing the non-pathogenic IB phylotype. This formulation A is therefore capable of rebalancing the C. acnes phylotypes in non-lesional areas of subjects with mild to moderate acne.

[0136] All of these results demonstrate that prolonged application of formulation A not only allows an improvement in acne on the lesional areas, but also on the non-lesional areas still called subclinical, i.e. areas where acne is not yet visible. Such a formulation therefore has the advantage of being applied in the context of acne prevention in subjects with acne-prone skin.

Claims

Claims

1. Cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, with at least one cosmetically acceptable or dermatologically acceptable excipient, for use in the treatment and / or prevention of acne in non-lesional areas of acne-prone skin.

2. Cosmetic or dermatological composition comprising a myrtle extract and a Tripterygium wilfordii extract, with at least one cosmetically acceptable or dermatologically acceptable excipient, for use in the prevention of acne.

3. Composition for its use according to claim 1 or 2, characterized in that the myrtle extract is an apolar fraction comprising myrtucommulones and ursolic acid.

4. Composition for use according to claim 3, characterized in that the myrtucommulones comprise myrtucommulones A, B', D, B, isosemimyrtucommulone and semimyrtucommulone.

5. Composition for its use according to claim 3 or 4, characterized in that the content of total myrtucommulones is between 3% and 10% by weight, relative to the total weight of the dry myrtle extract.

6. Composition for its use according to any one of claims 3 to 5, characterized in that the ursolic acid content is between 10% and 30% by weight, relative to the total weight of the dry myrtle extract.

7. Composition for its use according to any one of claims 1 to 6, characterized in that the extract of Tripterygium wilfordii comprises at least one pentacyclic triterpene.

8. Composition for its use according to claim 7, characterized in that the at least one pentacyclic triterpene is chosen from Tingenin A, Tingenin B, Celastrol, Pristimerine, Trip-terygone and their mixtures.

9. Composition for its use according to claim 8, characterized in that the at least one pentacyclic triterpene is chosen from Tingenin A, Tingenin B, Celastrol, and their mixtures, and preferably is a mixture of Tingenin A, Tingenin B, and Celastrol.

10. Composition for its use according to any one of claims 7 to 9, characterized in that the extract of Tripterygium wilfordii is capable of being obtained by the following method: (i) A proliferation phase of Tripterygium wilfordii cells in a proliferation medium, (ii) An elicitation phase by adding an elicitation cocktail to the cell culture obtained in step (i), said elicitation cocktail comprising at least one elicitor of monocarboxylic compound type and at least one biotic elicitor, and (iii) The preparation of an extract comprising pentacyclic triterpenes from the cell culture obtained in step (ii).

11. Composition for use according to any one of claims 1 to 10, characterized in that the extract of Tripterygium wilfordii comprises 90% by weight of at least one pentacyclic triterpene relative to the total weight of the dry extract.

12. Composition for use according to any one of claims 1 to 11, characterized in that the extract of Tripterygium wilfordii comprises between 1% and 20% of Tingenin A by weight relative to the total weight of the dry extract, between 1% and 20% of Tingenin B by weight relative to the weight of the dry extract, and at least 60% of Celastrol by weight relative to the total weight of the dry extract.

13. Composition for use according to any one of claims 1 to 12, characterized in that it comprises 0.01% to 1% of dry extract of Tripterygium wilfordii by weight relative to the total weight of the composition.

14. Composition for use according to any one of claims 1 to 13, characterized in that it comprises 0.01% to 1% by weight of a dry extract of myrtle relative to the total weight of the composition.

15. Composition for use according to any one of claims 1 to 14, characterized in that it is in a form suitable for topical application.

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