Cosmetic use of the oily extract of the solid residue of the immortelle as a skin lightening agent

Abstract

The present invention relates to the cosmetic use of an oily extract of the solid residues of preserved flowers as a skin lightening agent. It also relates to a cosmetic composition comprising, in a physiologically acceptable medium, an oily extract of the residues of preserved flowers and hexylresorcinol; and the use of this composition.

Description

Technical Field

[0001] This invention relates to the cosmetic use of an oily extract of preserved flower distillation residues as a skin brightening agent. The invention also relates to cosmetic compositions comprising an oily extract of preserved flower distillation residues and hexylresorcinol in a physiologically acceptable medium, and further relates to the use of such compositions. Background Technology

[0002] The mechanism of skin pigmentation involves a complex process of melanin production by melanocytes, specialized cells located in the basal layer of the epidermis. Illustrated, this mechanism involves the following main steps:

[0003] Tyrosine -> Dopa -> Dopaquinone -> Dopachrome -> Melanin

[0004] Catalyzed by tyrosinase, a key enzyme involved in the reaction sequence.

[0005] Skin color is primarily determined by the type and concentration of melanin produced by melanocytes. Melanocytes can be activated by internal or external stimuli to produce melanin in specialized lysosomal-like organelles called melanosomes. Upon maturation, perinuclear melanosomes are transported along microtubules and the actin cytoskeleton to the periphery of melanocyte dendrites. Melanin-containing melanosomes are then transferred to adjacent keratinocytes.

[0006] Damage to any step in this complex process can lead to skin pigmentation disorders. In particular, an increase in the number of active melanocytes and / or an increase in melanin production and / or an increase in the transfer of melanosomes from melanocytes to keratinocytes may lead to hyperpigmentation of the skin.

[0007] Therefore, environmental, genetic, and / or hormonal factors can regulate the amount, type, and distribution of melanin in the skin and / or hair, with significant cosmetic and psychological consequences. Thus, darker and / or more pigmentation may appear on the skin under the sun, more specifically on the hands of some individuals. Other types of regional hyperpigmentation due to excessive melanocyte activity are idiopathic melasma that occur during pregnancy (“pregnancy mask” or melasma) or during estrogen-progesterone contraception. Conversely, hypopigmentation can be observed in cases of scarring after sun exposure, or in certain vitiligo lesions such as vitiligo. In these cases, if it is not possible to re-pigment the damaged skin, the remaining areas of normal skin are depigmented to give the skin a uniform white tone as a whole.

[0008] There is a particular need to use harmless, topical depigmenting substances with good efficacy to treat all these hyperpigmentation issues. A substance is considered bleaching or depigmenting if it acts directly on the activity of epidermal melanocytes where melanin production occurs, and / or if it interferes with one of the steps in melanin biosynthesis, by inhibiting one of the enzymes involved in melanin production, particularly tyrosinase, or by a structural analog of a chemical compound embedded in the melanin synthesis chain, thus blocking that chain and ensuring depigmentation and therefore skin brightening.

[0009] Various cosmetic active ingredients, such as vitamin C and its derivatives, arbutin, azelaic acid, glycolic acid, butylresorcinol (or rutin), hexylresorcinol, and glutathione and its derivatives, are currently used to treat these hyperpigmentation problems. These compounds have different modes of action, including inhibiting tyrosinase activity, reducing the amount of melanin formed, and even stimulating melanin elimination in keratinocytes. However, some of these compounds may cause side effects such as burns, erythema, and dry skin.

[0010] Therefore, there is still a need for a new type of human skin bleach that is as effective as those known skin bleachs, but without their drawbacks, namely, that it is non-irritating, non-toxic, and / or non-allergenic to the skin.

[0011] In this context, the use of various preserved flower extracts as bleaching agents has been proposed, particularly extracts obtained from the aerial parts of *Helichrysum gymnocephalum (DC) Humbert* using solvents such as ethanol. These extracts are characterized by the presence of monoterpene derivatives of chalcone or dihydrochalcone, particularly gymnochalcone (FR2970416). Reference can also be made to WO 2007 / 015232, which suggests the use of an (undefined) extract of *Helichrysum arenarium* in the treatment of scars and skin pigmentation, and CN111973544, which discloses the depigmenting effects of a water-diol extract of *Helichrysum bracteatum*. Bleaching properties have also been found in the essential oil of *Helichrysum angustifolium* (CN108670926). In addition, combinations of several different preserved flower extracts have been proposed for their anti-aging properties and for their ability to reduce pigmentation spots (FR3003167). Finally, patent application WO 2019 / 086602 indicates that a subcritical water extract of the flowering top of Helichrysum italicum containing caffeoylquinic acid may have depigmenting properties and be used as an anti-blemish cosmetic active agent.

[0012] The applicant has now demonstrated, quite surprisingly, that another preserved flower extract known for its properties of stimulating keratinocyte differentiation and its ability to restore the skin barrier (WO 2019 / 180368) exhibits good depigmenting activity even at low concentrations, and is also well tolerated. Summary of the Invention

[0013] To be precise, the subject of this invention is the cosmetic use of an oily extract of the distillation residue of the Italian immortelle species, applied topically to the skin, as an agent for bleaching and / or brightening skin tone and / or homogenizing skin color and / or reducing or preventing the appearance of pigmentation spots.

[0014] Another subject of the invention is an agent for bleaching and / or brightening skin tone and / or for homogenizing skin color and / or reducing or preventing the appearance of pigmentation spots, said agent consisting of an oily extract of the distillation residue of the immortal flower of the Italian immortelle species.

[0015] Another subject of the invention relates to a cosmetic composition comprising, in a physiologically acceptable medium, 0.01 wt% to 1 wt% of an oily extract of preserved flower distillation residue as defined above, and 0.001 wt% to 5 wt% of hexylresorcinol, relative to the total weight of the composition.

[0016] The subject of this invention is also the cosmetic use of the above-mentioned cosmetic composition for bleaching skin and / or brightening skin tone and / or homogenizing skin color and / or reducing or preventing the appearance of pigmentation spots, including topical application of the composition to the skin.

[0017] The subject of this invention is also a cosmetic method for bleaching and / or brightening skin tone and / or homogenizing skin color and / or reducing or preventing the appearance of pigmentation spots, comprising applying the above composition topically to the skin. Detailed Implementation

[0018] The present invention uses an oily extract of the distillation residue of the preserved flowers of the Italian immortelle species, preferably the Italian immortelle species from Corsica.

[0019] The term "distillation residue" refers to the solid residue remaining after water distillation or steam distillation of any part of a preserved flower plant and separation of the resulting essential oils and hydrosols. Therefore, the preserved flower distillation residue used according to the invention is obtained by a method comprising the steps of water distillation or steam distillation of all or part of the plant, preferably the above-ground parts of the plant, particularly its flowers or flowering tops, followed by the recovery of the distillation or steam distillation residue.

[0020] An oily extract from the distillation residue can then be obtained by any method used for extracting oil from the distillation residue, and more specifically by using a supercritical fluid (e.g., carbon dioxide, nitric oxide, dimethyl ether, propane, ethylene, or methanol, this list is not limiting). Supercritical carbon dioxide can therefore be used. Those skilled in the art know how to adjust extraction parameters, particularly temperature, pressure, and fluid flow rate, according to the desired extraction rate and yield. For example, extraction using supercritical CO2 can be carried out at a pressure of 250 to 300 bar and a temperature of 40 to 80°C, preferably 50 to 70°C. After extraction, the supercritical fluid is returned to the gaseous state by depressurization and is usually recycled. The resulting plant extract is advantageously subjected to a molecular distillation step, for example at a temperature of 180-250°C, preferably 200-220°C, to concentrate unsaponifiable matter present in the extract. Prior to this molecular distillation step, the plant extract can optionally be diluted in an oily solvent, particularly an oily solvent based on vegetable oils such as sunflower oil. This oily solvent is removed during the molecular distillation step.

[0021] According to a preferred embodiment, the oily extract of the distillation residue of preserved flowers used in this invention is obtained by a method comprising the steps of: extracting the distillation residue with a supercritical fluid, preferably carbon dioxide in a supercritical state, and optionally a subsequent molecular distillation step.

[0022] The resulting distillate contains, in particular, 30% to 50% by weight, more particularly 35% to 45% by weight, of unsaponifiable matter and 1% to 10% by weight, more particularly 3% to 7% by weight, of sterols, primarily β-sitosterol.

[0023] Extracts obtained using supercritical fluids or distillates produced by molecular distillation steps may optionally be diluted in an oily solvent to a concentration of 1% to 10% by weight, for example, particularly 3% to 5% by weight, for ease of handling. Any oily solvent suitable for cosmetic purposes may be used for this purpose, particularly fatty acid triglycerides, such as caprylic / capric triglycerides.

[0024] The applicant characterized the oily extract of the distillation residue of preserved flowers by gas chromatography coupled to mass spectrometry. This demonstrates that, as determined by gas chromatography, the oily extract of the distillation residue of preserved flowers contains less than 10% volatile fractions.

[0025] It is also shown that the oily extract of the preserved flower distillation residue used according to the present invention differs in composition from the extract of the preserved flower flower top obtained by supercritical CO2 in that it substantially does not contain (i.e., less than 10% by weight, or even less than 5% by weight, or even less than 2% by weight) palmitoleic acid, linalool, nerol, 2-decenal, 2-tetane, heptadecane, 8-heptadecene, isobutyric acid, isovaleric acid, hexadecenol, lauric acid, pentadecanoic acid, decanoic acid, and isostearic acid. More generally, the extract does not contain saturated fatty acids containing more than 10 carbon atoms. In contrast, the oily extract of the preserved flower distillation residue used according to the present invention contains components not yet identified in the extract of the preserved flower flower top obtained by supercritical CO2, namely 2(Z)-decenoic acid and 4(Z)-decenoic acid.

[0026] Furthermore, compared to preserved flower essential oil, the oily extract of preserved flower distillation residue used according to the present invention contains sterols, essentially β-sitosterol, straight-chain carboxylic acids, and C7-C... 10 Straight-chain saturated or unsaturated carboxylic acids (especially heptanoic acid, octenic acid, and decenoic acid), benzoic acid, and C8-C... 18 Ethyl esters of straight-chain saturated or unsaturated fatty acids (especially caprylate, decanoate and linoleate).

[0027] It also differs from the preserved flower oil extract obtained by using sunflower oil extraction in that it does not contain any limonene and contains fatty acids and / or their ethyl esters (heptanoic acid, 2-octenic acid, 4-octenic acid, nonanoic acid, 2-decenoic acid, 4-decenoic acid, linoleic acid) and benzoic acid, which are not present in the above-mentioned oil extracts.

[0028] Finally, the oily extract of the preserved flower distillation residue used according to the present invention is low in polyphenols and, in particular, free of dicaffeoylquinic acid and chalcone.

[0029] The oily extract of the preserved flower distillation residue advantageously accounts for 0.01% to 1% relative to the total weight of the composition according to the invention.

[0030] The oily extract of the distillation residue of preserved flowers according to the invention is contained in a cosmetic composition containing a physiologically acceptable medium, particularly a cosmetically acceptable medium, i.e., a medium that does not cause any stinging or redness incompatible with cosmetic use. This medium preferably contains an aqueous phase and / or a fatty phase. Preferably, the composition is in emulsion form, particularly an oil-in-water emulsion or a water-in-oil emulsion, or an anhydrous oily composition. The term "anhydrous composition" refers to a composition containing less than 5% by weight of water, preferably less than 2% by weight of water, or even no water at all.

[0031] When present, the aqueous phase comprises water and optionally at least one component selected from polyols and aqueous gelling agents. Advantageously, water constitutes 40% to 80% of the total weight of the composition, for example, 50% to 70%. The polyol may be particularly selected from glycerol, propylene glycol, butylene glycol, pentanediol, and mixtures thereof, and may constitute 5% to 30% of the total weight of the composition, preferably 15% to 25%.

[0032] The term "aqueous gelling agent" refers to a polymeric compound capable of fixing water molecules through hydration and thus increasing the viscosity of the aqueous phase. Such gelling agents may be selected from: polysaccharides, such as cellulose and its derivatives, modified starch, carrageenan, agar, xanthan gum, and plant gums, such as guar gum or locust bean gum; synthetic polymers, particularly optionally crosslinked sodium acrylate homopolymers, and acrylic copolymers, particularly sodium acrylate and / or (meth)acrylate alkyl esters and / or (meth)acrylate hydroxyalkyl esters and / or (meth)acrylate (polyethoxy)alkyl ester copolymers, optionally having at least one other monomer, advantageously 2-acrylamido-2-methylpropanesulfonic acid (AMPS), these copolymers optionally crosslinked; and mixtures thereof.

[0033] For a portion thereof, when present, the aliphatic phase may contain one or more volatile and / or non-volatile oils. Examples of volatile oils are branched alkanes, such as isododecane, and straight-chain C64... 10 -C 13 Alkanes. Non-volatile oils that may be specifically mentioned include:

[0034] -Acids and monohydric alcohol esters, selected from: monoesters and polyesters of saturated linear C2-C10 (preferably C6-C10) acids and saturated linear C10-C18 (preferably C10-C14) monohydric alcohols; monoesters and polyesters of saturated linear C10-C20 acids and branched or unsaturated C3-C20 (preferably C3-C10) monohydric alcohols; monoesters and polyesters of branched or unsaturated C5-C20 acids and branched or unsaturated C5-C20 monohydric alcohols; and monoesters and polyesters of branched or unsaturated C5-C20 acids and straight-chain C2-C4 monohydric alcohols.

[0035] Triglycerides of -C6-C12 fatty acids, such as caprylic and capric triglycerides and triheptanoic triglycerides;

[0036] - Branched and / or unsaturated C10-C20 fatty acids (such as oleic acid and linoleic acid);

[0037] - Branched and / or unsaturated C10-C20 fatty alcohols (e.g., octyldodecanool and oleyl alcohol);

[0038] - Hydrocarbons, such as squalane (C30), especially plant-derived squalane extracted from olive oil, and hemisqualane (C15);

[0039] - Dialkyl carbonates, such as dioctyl carbonate and diethylhexyl carbonate;

[0040] -Dialkyl ethers, such as dioctyl ether; and

[0041] - Its mixture.

[0042] Vegetable oils containing one or more of the above-mentioned ingredients may also be mentioned.

[0043] Esters of acids and monohydric alcohols, particularly monoesters, may be mentioned, such as mixtures of cocoyl decanoate / caprylate, ethyl maleate, ethyl shea butter, isostearyl isostearate, isononyl isononanoate, ethylhexyl isononanoate, hexyl neopentanoate, ethylhexyl neopentanoate, isostearyl neopentanoate, isodecanyl neopentanoate, isopropyl myristate, octyl dodecyl myristate, isopropyl palmitate, ethylhexyl palmitate, hexyl laurate, isoamyl laurate, cetearyl nonanoate, propyl heptaethyl octanoate, and mixtures thereof. Other esters that may be used include diesters of acids and monohydric alcohols, such as diisopropyl adipate, diethylhexyl adipate, diisopropyl sebacate, and diisoamyl sebacate.

[0044] Examples of vegetable oils include, in particular, wheat germ oil, sunflower oil, argan oil, hibiscus oil, coriander oil, grapeseed oil, sesame oil, corn oil, almond oil, castor oil, shea butter, avocado oil, olive oil, soybean oil, sweet almond oil, palm oil, rapeseed oil, cottonseed oil, hazelnut oil, macadamia nut oil, jojoba oil, alfalfa oil, rice oil, pumpkin oil, squash oil, blackcurrant oil, evening primrose oil, lavender oil, borage oil, millet oil, barley oil, quinoa oil, rye oil, safflower oil, candelilla oil, passion fruit oil, rosehip oil, blue thistle oil, flaxseed oil, and camellia oil.

[0045] The fatty phase may also contain at least one fatty phase structuring agent. The term "fatty phase structuring agent" refers to a compound capable of thickening oils contained in the composition, particularly selected from waxes, fatty phase gelling agents, and pasty fatty substances, and mixtures thereof.

[0046] According to a preferred embodiment, the composition further comprises at least one depigmenting agent in addition to an oily extract of the distillation residue of immortal flowers, which may be particularly selected from: plant extracts, especially licorice extract; vitamin C and its derivatives, particularly ascorbic acid, ascorbate glucoside, ascorbate ethyl ether, ascorbate tetraisopalmitate and magnesium ascorbate phosphate; arbutin; ferulic acid; kojic acid; resorcinol and its derivatives, such as hexylresorcinol and 4-(1-phenylethyl)-1,3-benzenediol, particularly by Symrise under the trade name Symwhite. Sold; salicylic acid, its salts and esters; meadowsweet and iris flower (Iris florentina) extracts; dipotassium glycyrrhizate; tranexamic acid; ellagic acid; nicotinic acid derivatives, such as nicotinamide; kiwi (Actinidia chinensis) extract sold by Gattefossé; by Ichimaru Pharcos under the trade name Liquid sells peony root (Paeonia suffruticosa) extract; marketed by SEPPIC under the brand name... The sale of lipid amino acids (phenylalanine modified with undecenoic acid); Italian immortelle essential oil; and mixtures thereof.

[0047] In a preferred embodiment of the invention, the cosmetic composition contains, in a physiologically acceptable medium, 0.01% to 1% by weight, preferably 0.05% to 1% by weight, more preferably 0.05% to 0.3% by weight of an oily extract of preserved flower distillation residue as defined above, and 0.001% to 5% by weight of hexylresorcinol, relative to the total weight of the composition.

[0048] Hexylresorcinol or 4-hexyl-1,3-benzenediol refers to compounds with the following chemical structures:

[0049]

[0050] Hexylresorcinol is known for its depigmenting and skin-care properties. Preferably, the hexylresorcinol introduced into the cosmetic compositions of the present invention is purified and virtually free of resorcinol (the latter compound is unstable and irritating to the skin). For this purpose, products obtained via the method described in Sythéon patent EP 2152 685 can be used, or commercially available products known to contain more than 99.5% hexylresorcinol can be used directly. HR, to inhibit melanin production in vitro (Funasaka AH et al., J.Lipid Res. 1999) and has bleaching activity on human volunteer skin (EP 2152685).

[0051] As shown in the examples below, the oily extract of the preserved flower distillation residue according to the present invention has been shown to enhance the depigmenting effect of hexylresorcinol.

[0052] As a variant or additional, the cosmetic composition may contain at least one active agent selected from: free radical scavengers, moisturizers, agents for stimulating the differentiation and / or proliferation of keratinocytes and / or fibroblasts; agents for stimulating the synthesis of glycosaminoglycans and / or collagen and / or dermal anchoring fibrils and / or elastic fibers; agents for preventing the degradation of collagen and / or glycosaminoglycans and / or dermal anchoring fibrils and / or elastic fibers; anti-glycation agents; and mixtures thereof.

[0053] Examples of such anti-aging active agents include, in particular: ascorbic acid, and its salts, ethers, and esters, especially ascorbate glucoside; adenosine; ribose; honey extracts; proteins and glycoproteins, especially those extracted from sweet almonds; hydrolyzed plant proteins, especially those derived from rice, hibiscus seeds, or lupins; polypeptides and pseudo-dipeptides, such as decarboxylated carnosine hydrochloride, respectively, under trade names. 3000 and Synthe's '6 sells palmitoyl pentapeptide-4 (Pal-Lys-Thr-Thr-Lys-Ser) and palmitoyl tripeptide-38, as well as products marketed by Lucas Meyer under the trade name... Palmitoyl tripeptide-8 is sold by Lipotec under the brand name... The pentapeptide-18 sold by Solution is marketed by Sandream under the brand name... And by Infinitec under the trade name X50 Powder sells palmitoyl hexapeptide-52; silanes, such as methylsilanol mannouronic acid; particularly arabinoxylan extracted from rye flour and particularly arabinogalactan from larch; hyaluronic acid and its salts; polyphenols, particularly those extracted from mimosa; α-hydroxy acids, including those extracted from lemon and those produced by Naturex under the trade name Hibiscus. Those sold; plant extracts (usually aqueous), such as alfalfa, wild pansy, horsetail, purslane (Acmella oleracea), large-winged thistle (Onopordum acanthium), yarrow (Achillea millefolium), especially included in products from BASF. (Chinese), *Embelia concinna* (sold by SEPPIC), and *Opuntia ficus indica* (specially marketed by Mibelle AG Biochemistry under the trade name) Sage (Salvia officinalis, especially sold by Provital Group), Vitex negundo (by Laboratoires Expanscience under the trade name) (sales), chestnuts, papayas, argan, oats, sunflowers, daisies, peonies, or dill; aquatic algae extracts, especially coral, slender-beaded coralweed, wakame, alaria esculenta, or nannochlorosis oculata; essential oils, especially myrtle essential oil; zinc and / or copper gluconate; and mixtures thereof.

[0054] The compositions according to the invention may also contain various components that are dispersible in the fatty phase and / or in the aqueous phase, provided that these components are compatible with topical application to the skin.

[0055] Therefore, it may contain at least one typically nonionic oil-in-water or water-in-oil emulsifier, such as polyoxyethylene ester, optionally polyethoxylated sorbitol ester, optionally polyethoxylated fatty acid ester of glycerol, fatty alcohol ether of sugar, such as alkyl glucoside, and mixtures thereof. The emulsifier may comprise 2% to 10% of the total weight of the composition, preferably 4% to 6%.

[0056] The compositions according to the invention may also comprise one or more powdered fillers, which are advantageously porous or hollow, preferably in the form of porous particles. These particles are, in principle, substantially spherical. These fillers may be particularly selected from:

[0057] - Organic fillers, such as: powders of polysaccharides, especially natural starch, modified starch or cellulose; powders of acrylic polymers such as poly(methyl methacrylate), polyamide or polyolefin; powders of dried algae such as coral algae (Corallina officinalis);

[0058] - Mineral fillers such as silica, clay, perlite and talc;

[0059] - and its mixtures.

[0060] Silica is preferred as a mineral filler.

[0061] These fillers can comprise 1% to 5% by weight of the total composition.

[0062] The compositions according to the invention may further comprise additives, particularly selected from organic and / or inorganic photoprotectants, which are active in blue light and / or UVA and / or UVB; and polysaccharide-based film-forming polymers capable of forming antifouling protective films, such as those produced by Solabia under the trade name... and Products for sale; fragrances; antioxidants; multivalent chelating agents; pH adjusters; preservatives; pigments; colorings; and mixtures thereof.

[0063] The composition can be in any form suitable for topical application to the skin, particularly in the form of lotion, cream, fluid, lotion, gel, paste, oil, or film. It is typically a no-rinse composition, especially a skin care composition.

[0064] As a variation, the composition according to the invention can be a wash-off composition for skin care, particularly for the face and possibly the body. In this case, it can be used as, for example, a face mask or an exfoliating paste.

[0065] The compositions according to the invention can be applied to the skin of individuals with skin discoloration mediated by environmental factors, particularly pigmentation spots, melasma, and / or pigmentation defects due to scars or acne marks. They are typically applied to at least one area of ​​the relevant body, and more specifically, to the skin of the face, collarbone, neck, arms, hands, and / or legs, with the aim of brightening these areas and / or unifying skin color. The compositions can be applied once or more daily, for example, in the morning and / or evening.

[0066] In any case, apply an oily extract of the distillation residue of immortal flower to the skin in an amount sufficient to inhibit melanin production. For example, 0.5 to 5 mg / cm³ can be used. 2 A cosmetic composition is applied daily to the relevant skin area, the cosmetic composition containing 0.01% to 1% by weight of an oily extract of the preserved flower distillation residue according to the invention. Attached Figure Description

[0067] Figure 1 This demonstrates the inhibitory effect of the extract according to the present invention on melanin production in mouse B16 melanocytes compared to two positive controls.

[0068] Figure 2 This demonstrates the inhibitory effect of the oily extract of Helichrysum itinerary on melanin production in mouse B16 melanocytes compared to two positive controls.

[0069] Figure 3 This demonstrates the inhibitory effect of the extract according to the invention in combination with hexylresorcinol on melanin production in mouse B16 melanocytes compared to each of the two active agents and two positive controls.

[0070] Example

[0071] The invention will be more clearly understood based on the following embodiments, which are given by way of illustration only and are not intended to limit the scope of the invention as defined by the claims.

[0072] Example 1 Preparation and characterization of extracts - comparison with oily extracts from flower tips

[0073] 1.1 Preparation of Extracts

[0074] Essential oils and hydrosols were produced by water distillation of the flowering tops of Italian immortelle. The water distillation residue (distillation residue) was then separated and extracted using supercritical CO2 at 60°C and 285 bar. The resulting unsaponifiable matter was diluted in sunflower oil and then subjected to molecular distillation at 200°C. The distillate was normalized by diluting the resulting fraction with caprylic / capric triglycerides to 3-5% by weight.

[0075] The polyphenol composition of this extract was compared with that of the following polyphenol composition obtained from the oily extract of Helichrysum italicum: 9 wt% of dried and ground plant powder (flower tops) was mixed with 91% ultrasonically deodorized sunflower oil in a container cooled to 10°C. Extraction was carried out for 30 minutes under ultrasonication with stirring at 500 rpm. Microwave extraction was continued under a nitrogen flow of 0.4 L / min. The obtained solid and liquid oily phases were separated by decantation, and the oily phase was then filtered and stored under an inert atmosphere, protected from light.

[0076] Characterization of 1,2-polyphenols

[0077] Material

[0078] -Ultra-high performance liquid chromatograph (Agilent Technologies, USA)

[0079] -1290 tandem automatic injector

[0080] -1260 Series Diode Array Detector (DAD)

[0081] - Mass spectrometer: Equire 6000 (Brüker Daltonics, Bremen) equipped with an electroinjection ionization (ESI) source.

[0082] Sample preparation

[0083] The extract according to the invention was prepared by dissolving 2 g of the distillation residue extract in 1 mL of hexane. The oily solution was extracted three times with 2 mL of a MeOH / H2O mixture (60 / 40). Then the MeOH / H2O phases were combined and diluted 1 / 2 in a MeOH / H2O mixture (60 / 40).

[0084] Analytical method

[0085] The solution obtained after dilution / extraction was filtered through a PTFE filter (0.45 μm) and then injected (1 μL) onto an Agilent C18 column (2.1 mm x 100 mm; 1.8 μm) at a temperature maintained at 25 °C with a flow rate of 0.4 mL / min. The solvents used were: A = H2O / HCOOH (0.1%) and B = ACN / HCOOH (0.1%). The polyphenol elution gradient was as follows:

[0086] [Table 1]

[0087]

[0088]

[0089] After leaving the diode array detector, the eluate was injected into the mass spectrometer. The analysis was carried out in negative or positive mode.

[0090] LC-MS spectra were acquired over the entire mass range (m / z) from 100 to 1400. Then all data were collected and processed using Hystar version 3.0 software.

[0091] Results

[0092] [Table 2]

[0093]

[0094]

[0095] n.d: not detected; < LQ: detected but below the limit of quantification

[0096] Characterization of 1,3-chalcone

[0097] The presence of four chalcones (linderatin, linderachalcone, methyl-linderatin, gymnochalcone) in the distillation residue extract described above was evaluated by using 2D NMR method with 1 H / 13 C HSQC spectra.

[0098] Materials and Methods: Aliquots (15 mg) of the distillation residue extract were dissolved in 700 μL DMSO-d6 for analysis by 2D NMR at 298 Kelvin.

[0099] Use equipped with 2DNMR analysis was performed using a Brüker Avance AVIII-600 spectrometer with an NMR probe. The obtained NMR values ​​were compared with those calculated using ACD Labs software.

[0100] Result: Chalcone was not detected in the sample.

[0101] 1.4 Characterization of other components

[0102] In parallel, the presence of various organic compounds was evaluated in both of the above-mentioned extracts, and also in extracts obtained in a manner similar to that of the extracts according to the invention, but using a process performed on the flower heads of preserved flowers rather than on their water distillation residues.

[0103] Material

[0104] - Micro-GC Ultragas Chromatograph (ThermoElectron SAS)

[0105] - Mass spectrometer: El LT Large T ISQ mass spectrometer (ThermoElectron SAS) used in E1 mode.

[0106] Sample preparation

[0107] The sample was prepared by diluting 0.3 g of the extract in 6 mL of isooctane. 0.6 mL of methyl methacrylate (MDMA) was added, and the mixture was vortexed for 30 seconds.

[0108] Add 1g NaHSO4 and vortex the mixture for 30 seconds.

[0109] The mixture is then separated by sedimentation.

[0110] The supernatant was collected for chromatographic analysis.

[0111] Analytical methods

[0112] Analyze the sample under the following conditions:

[0113] TR WAX 60m x 0.25μm x 0.25mm polar chromatographic column

[0114] H2 flow rate: 1 mL / min

[0115] Oven program: 50℃ (2 minutes), then 10℃ / minute until 250℃ (18 minutes).

[0116] Separation ratio: 100

[0117] Injection volume: 1 μL

[0118] Syringe temperature: 250℃

[0119] Quality range: 50-650 amu

[0120] result

[0121] It is worth noting that the extract of the preserved flower distillation residue according to the present invention differs from the oily extract of the flower top in that it does not contain limonene, contains significantly more β-eudesmol, and contains fatty acids and / or their ethyl esters (heptanoic acid, 2-octenic acid, 4-octenic acid, nonanoic acid, 2-decenoic acid, 4-decenoic acid, and linoleic acid) and benzoic acid, which are not present in the oily extract of the preserved flower flower top.

[0122] Furthermore, the distillation residue extract according to the invention differs significantly from the supercritical CO2 extract from the flower apex because it does not contain saturated fatty acids containing more than 10 carbon atoms, and differs in that it contains far fewer (if any) terpenoids (especially monoterpenes and sesquiterpenes), particularly nerol and linalool, and far fewer (if any) heavy alkanes (containing more than 16 carbon atoms), and is free of palmitoleic acid, isobutyric acid and isovaleric acid, 2-decenal and hexadecenol. In addition, the distillation residue extract according to the invention contains characteristic molecules not present in the supercritical CO2 extract from the flower apex, namely (2)-(Z)-decenoic acid and 4-(Z)-decenoic acid.

[0123] Example 2 The effect of the preserved flower distillation residue extract on melanin production according to the present invention

[0124] The preserved flower distillation residue extract and the oily extract from the flower tops described in Example 1 were tested to evaluate their ability to inhibit melanin production.

[0125] 2,1-Materials and Methods

[0126] Melanocytes (mouse B16 cell line) were seeded at 2000 cells / well in phenol red-free DMEM medium containing 10% fetal bovine serum, 2 mM glutamine, 1 mM sodium pyruvate, and 100 μg / ml anti-viscosin in 96-well plates. The next day, cells were treated with 0.1 μM NDP-MSH to induce pigmentation. Under the test conditions, 3 mM kojic acid and 0.5% vitamin C (ascorbic acid glucoside) were used as positive controls for pigmentation inhibition in the presence of NDP-MSH, and the extract to be evaluated was tested at several concentrations. All conditions were tested in triplicate. Cells were incubated at 37°C and 5% CO2 for 72 h in the presence of the extract and the positive control. Cell viability was then assessed using the XTT method, and only non-cytotoxic conditions with melanin levels (whose viability was greater than 85% of that of the untreated control) were analyzed.

[0127] For melanin assay, culture medium was aspirated from each well, and cells were lysed by adding 50 μl of 1M NaOH to the cell population. The plate was shaken at room temperature for 30 minutes, and then the lysate was transferred to a glass flask and incubated overnight at 95°C. After incubation, the lysate was centrifuged at 2000 rpm to remove cell debris. The absorbance of the supernatant was read at 405 nm, reflecting the amount of melanin in each well, and the average of three replicates was determined for each condition. Wells containing cells not treated with NDP-MSH served as controls, and their OD values ​​were subtracted from all OD values ​​of the other wells. The percentage of melanin inhibition was then calculated according to the following formula:

[0128] 100 - ((Melanin amount in test conditions / Melanin amount in untreated control) x 100)

[0129] The significance of the results was assessed using a one-way Anova test (for distillation residue extracts and oily extracts from flower tips) or a Student's test (for kojic acid and vitamin C).

[0130] 2.2 Results

[0131] like Figure 1 and 2 As shown, the efficacy of kojic acid and vitamin C was verified by testing (***p<0.001; ****p<0.0001).

[0132] Figure 1It is also shown that the preserved flower distillation residue extract according to the invention significantly inhibits melanocyte pigmentation in a dose-dependent manner at concentrations of 0.05% or higher (0.1%, p < 0.0001; 0.05%, p < 0.05). At a concentration of 0.1%, the distillation residue extract according to the invention exhibits significantly higher efficacy than the highest concentration of vitamin C, which constitutes a bleaching active agent commonly used in cosmetics. Its efficacy is close to that of kojic acid as a benchmark depigmenting agent, but kojic acid is known to involve serious risks of allergies and irritation.

[0133] Therefore, this embodiment demonstrates that the preserved flower distillation residue extract according to the present invention constitutes a selection of cosmetic active agents for skin brightening.

[0134] In comparison, Figure 2 The study showed that the effect of the oily extract from the top of the preserved flower flower on pigmentation was not significantly different from that of the control.

[0135] Example 3 The effect of the combination of hexylresorcinol and the preserved flower distillation residue extract according to the present invention on melanin production.

[0136] The tests described in Example 2 were repeated, with hexylresorcinol added to the preserved flower distillation residue extract according to the invention. Preliminary cytotoxicity studies defined the maximum non-cytotoxic dose that could be administered to cells as 10 μg / mL of hexylresorcinol and 1% of the preserved flower distillation residue extract according to the invention. These two active agents were tested at concentrations of 0.035% preserved flower distillation residue extract and 5 μg / mL of hexylresorcinol.

[0137] The results were compared with those obtained by using the extract according to the invention alone and by using hexylresorcinol alone, at the same dosage.

[0138] like Figure 3 As shown, the efficacy of kojic acid and vitamin C (****p<0.0001) and the depigmenting efficacy of hexylresorcinol were verified by the test.

[0139] Furthermore, the tested combination inhibited pigmentation more strongly than when either of the two surfactants was used alone. This efficacy was not only significantly different from the untreated control, but also significantly different from each surfactant used alone at the same concentration (“one-way Anova test”, $$p<0.01 vs. distillate residue extract; Δp<0.05 vs. hexylresorcinol).

[0140] Therefore, this test demonstrates that the effect of hexylresorcinol on melanin production is significantly enhanced by the addition of the extract according to the present invention, which is unexpected.

[0141] Example 4 Cosmetic Compositions

[0142] The following composition is prepared in a manner conventional to those skilled in the art by mixing the following ingredients in the indicated weight proportions.

[0143] hand cream

[0144]

[0145]

[0146] Face oil and body oil Oil

Claims

1. A cosmetic method for bleaching skin and / or brightening skin tone and / or homogenizing skin color and / or reducing the appearance of pigmentation spots, said method comprising applying Italian immortelle ( Helichrysum italicum An oily extract of the distillation residue of the aerial parts of a species of immortal flower is applied topically to the skin, wherein the oily extract of the distillation residue is obtained according to a method comprising the following steps: extracting the distillation residue with carbon dioxide in a supercritical state.

2. The cosmetic method according to claim 1, characterized in that, As determined by gas chromatography, the oily extract of the preserved flower distillation residue contains less than 10% volatile fraction.

3. The cosmetic method according to claim 1 or 2, characterized in that, The oily extract of the preserved flower distillation residue contains linear saturated or unsaturated C7-C chains. 10 Carboxylic acids; and straight-chain saturated or unsaturated C8-C... 18 Ethyl esters of fatty acids.

4. The cosmetic method according to claim 3, characterized in that, The straight-chain saturated or unsaturated C7-C 10 Carboxylic acids include 2-(Z)-decenoic acid and 4-(Z)-decenoic acid.

5. The cosmetic method according to claim 3, characterized in that, The straight-chain saturated or unsaturated C7-C 10 Carboxylic acids include heptanoic acid, octenic acid, and decenoic acid; and the straight-chain saturated or unsaturated C8-C... 18 Ethyl esters of fatty acids include ethyl octanoate, ethyl decanoate, and ethyl linoleate.

6. The cosmetic method according to claim 1 or 2, characterized in that, The oily extract of the preserved flower distillation residue contains sterols.

7. The cosmetic method according to claim 6, characterized in that, The sterols are composed of β-sitosterol.

8. The cosmetic method according to claim 1 or 2, characterized in that, The preserved flowers of the Italian immortelle species are of Corsican origin.

9. The cosmetic method according to claim 1 or 2, characterized in that, The step of extracting the distillation residue with supercritical carbon dioxide is followed by a molecular distillation step.

10. The cosmetic method according to claim 9, characterized in that, The oily extract of the preserved flower distillation residue is diluted in an oily solvent.

11. The cosmetic method according to claim 10, characterized in that, The oily solvent is caprylic / capric triglyceride.

12. The cosmetic method according to claim 9, characterized in that, The preserved flower distillation residue is obtained according to a method comprising the following steps: a step of water distillation or steam distillation of the flower or the top of the flower, followed by a step of recovering the distillation or steam distillation residue.

13. The cosmetic method according to claim 1 or 2, characterized in that, The oily extract of the preserved flower distillation residue is applied to the skin of the face, collar, neck, arms, hands and / or legs.

14. The cosmetic method according to claim 1 or 2, characterized in that, The oily extract of the preserved flower distillation residue is applied to the skin of people with skin discoloration mediated by environmental factors.

15. A cosmetic composition comprising, in a physiologically acceptable medium, 0.01% to 1% by weight of an oily extract of an immortal flower distillation residue as defined in any one of claims 1 to 12, and 0.001% to 5% by weight of hexylresorcinol, relative to the total weight of the composition.

16. An agent for bleaching and / or brightening skin tone and / or homogenizing skin color and / or reducing the appearance of pigmentation spots, said agent comprising an oily extract of the distillation residue of the immortal flower of the Helichrysum tectorum species as defined in any one of claims 1 to 12.

17. A cosmetic method for bleaching and / or brightening skin tone and / or homogenizing skin color and / or reducing the appearance of pigmentation spots, said method comprising applying the composition of claim 15 topically to the skin.

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