Method for obtaining sandalwood extract, composition containing the same and cosmetic use thereof

Abstract

The present invention relates to a process for obtaining a sandalwood (Santalum album) extract by extraction using a supercritical fluid and a cosolvent selected from primary or secondary alcohols. The present invention also relates to a crude sandalwood extract comprising 10 to 70% of volatile compounds and 30 to 90% of semi-volatile to non-volatile compounds. The present invention also relates to a cosmetic composition comprising said extract in solubilized form and the cosmetic use of said composition for skin care, scalp care and skin appendage care.

Description

Technical Field

[0001] The present invention relates to the field of cosmetics and more specifically to the field of active ingredients used in the formulation of skin care compositions. The present invention relates to an extract of sandalwood (Santalum album) comprising from 10 to 70% of volatile compounds and from 30 to 90% of semi-volatile to non-volatile compounds, to a process for obtaining such an extract, to a cosmetic composition comprising it and finally to the cosmetic use of said composition for the care of the skin, scalp and appendages. Background Art

[0002] Santalum alum L., also known as Indian sandalwood, is a shrub of the Sandalaceae family. It is native to the Indian Peninsula and Southeast Asia and has been cultivated in Australia for a long time.

[0003] Sandalwood is used primarily in perfumery and cosmetics as an essential oil produced by distillation of the trunk and other parts of the tree (twigs, roots) or by more sophisticated extraction techniques.

[0004] The main compounds in sandalwood essential oil are sesquiterpene alcohols (content exceeding 90%), especially (Z)-α-santalol (not more than 50%) and (Z)-β-santalol (not more than 30%). (Z)-α-santalol has a woody cedarwood-like odor, while (Z)-β-santalol gives the characteristic fragrance of sandalwood (Brocke, C.; Eh, M.; Finke, A., Recent Developments in the Chemistry of Sandalwood Odorants, Chem. Biodiv. 2008, 5, 1000-1010).

[0005] Sandalwood essential oil contains other sesquiterpene alcohols, such as (E)-β-curcumin-12-ol (2%), (Z)-γ-bisabolol-12-ol (2%), (6R,7S)-iso-β-bisabolol (1%), (6S,7S)-iso-β-bisabolol, epi-α-bisabolol and (Z)-α-trans-bergamotol (6%) (Baldovini, N.; Delasalle, C.; Joulain, D., Phytochemistry of the heartwood from fragrant Santalum species: a review, Flavour Fragr. J. 2011, 26, 7-26).

[0006] Sandalwood is known and used in Ayurvedic medicine and aromatherapy to treat and prevent a variety of diseases, especially to relieve anxiety, stress and depression (Kumar, R.; Anjum, N.; Tripathi, JC, Phytochemistry and Pharmacology of Santalum album L.: A Review, World J. Pharm. Res. 2015, 4, 1842-1876; Setzer, WN, Essential Oils and Anxiolytic Aromatherapy, Nat. Prod. Commun. 2009, 4, 1305-1316).

[0007] Sandalwood - especially through the presence of the sesquiterpene α-santalol - is also known not only for its anti-inflammatory, antioxidant, antiviral and antibacterial activities, but also for its chemopreventive and anticancer properties (Santha, S.; Dwivedi, C. Anticancer effects of sandalwood (Santalum album), Anticancer Res. 2015, 35, 3137-3146).

[0008] The method of extracting sandalwood using supercritical CO2 is described in several documents. Document CN104232308 describes a method for extracting white sandalwood roots. After freeze drying and pulverization and grinding, the plant material is extracted with supercritical CO2 at a pressure of 30 to 35 MPa and a temperature of 50 to 65°C. Only the first two fractions are considered and combined to obtain the sandalwood extract.

[0009] It is also known from the work of Falconieri D. and colleagues that the recommended conditions for the extraction of plant materials by supercritical CO2 are: pressure 90 bar, temperature 50°C (density CO2 = 0.287 g / cm 3 ), CO2 flow rate is 0.6 to 1.5 Kg / h (Extraction of Essential Oils from Natural Matrices, Acta Hortic. 2010 4 229-240).

[0010] Marongiu B. et al. describe a method for obtaining essential oil from coarsely ground white sandalwood by supercritical CO2, using the following parameters: extraction temperature and pressure were 45°C and 120 bar (CO2 density = 0.658 g / cm 3), the separator temperature and pressure were 15°C and 20 bar respectively, and the flow rate of CO2 was fixed at 1.5 Kg / h. These conditions made it possible to obtain a sandalwood essential oil with a yield of 1.9%, which was mainly composed of α-santalol (46.1%), β-santalol (20.4%), epi-β-santalol (6.8%) and trans-α-bergamotol (5.4%). It was compared with the composition of the essential oil obtained by simple steam distillation (Extraction of Santalum album and Boswellia carteriiBirdw.volatile oil by supercritical carbon dioxide: influence of some process parameters Flavour Fragr.J.2006 21 718-724).

[0011] However, the sandalwood extracts described in the prior art have a characteristic olfactory note and contain a non-negligible amount of high-risk, volatile compounds, such as allergens. Since the raw materials used to produce the extract of the present invention have been consumed due to steam distillation, the developed extract does not contain volatile compounds and therefore allergens, and has an olfactory note of much lower intensity.

[0012] The extract of the present invention differs from the classically described sandalwood essential oil in that it contains primarily semi-volatile to non-volatile compounds as detailed below:

[0013] - phenolic compounds represented by mixtures of phenolic acids and aldehydes,

[0014] - lignans,

[0015] - and more non-polar compounds represented mainly by mixtures of acetylenic acids and fatty acids,

[0016] - sesquiterpene derivatives of sandalwood, such as dimers and aliphatic esters of sandalwood, and

[0017] -Saponifiable compounds.

[0018] Sandalwood extract is also known to work on the skin, particularly in preventing the appearance of wrinkles and fine lines (KR101220903B1). The essential oil is also recommended for the care of dry or irritated skin, skin inflammation or itchy scalp.

[0019] The skin is an organ composed of several layers (dermis, epidermis and stratum corneum) that covers the entire human body surface and ensures protective functions against external aggressions, sensory, immune, metabolic, thermoregulatory or even barrier functions, thereby limiting dehydration. In particular, when the skin is subjected to external aggressions, it has the ability to regenerate its barrier function, as well as antioxidant and detoxification defense mechanisms that are induced in response to these aggressions. Maintaining the barrier function of the skin is essential to maintaining its integrity.

[0020] The appearance of the skin can be altered by internal changes (intrinsic aging, disease, and hormonal changes such as pregnancy) or external factors (environmental factors such as pollution, sunlight, pathogens, temperature changes, etc.). All of these changes affect not only the skin, but also the keratinous appendages such as hair, eyelashes, eyebrows, nails and hair.

[0021] The olfactory receptor system in the skin (including OR2AT4) has an epidermal cell regeneration effect, including stimulating epidermal cell renewal and migration (Denda M. Newly discovered olfactory receptors in epidermal keratinocytes are associated with proliferation, migration, and re-epithelialisation of keratinocytes. J Invest Dermatol. 2014 Nov; 134(11): 2677-2679).

[0022] The skin and its appendages (hair follicles, sebaceous glands) have olfactory receptors, including the OR2AT4 receptor associated with hair growth (Chéret J. et al. Olfactory receptor OR2AT4 regulates human hair growth. Nat Commun 9, 3624, 2018).

[0023] New molecules that act on olfactory receptors are thought to play a role in the skin's ability to regenerate and detect external pollutants.

[0024] The inventors have thus demonstrated that a specific novel sandalwood extract, obtained from depleted sandalwood by an extraction process allowing the extraction of both volatile compounds and semi-volatile to non-volatile compounds of variable polarity (phenolic and lipid compounds), has beneficial effects on the skin and hair.

[0025] It has now been shown that, due to its specific composition, sandalwood extract has a higher biological activity than sandalwood essential oil. Summary of the invention

[0026] The present invention firstly relates to a method for obtaining a sandalwood (white sandalwood) extract, the method comprising the following steps:

[0027] a) adding 30 to 50% water to the consumed sandalwood;

[0028] b) optionally, mixing the wet sandalwood with 1 to 20% of an inert compound to increase the diffusion rate of the extraction solvent,

[0029] c) extracting using a supercritical fluid selected from carbon dioxide (CO2) and xenon in the presence of a polar cosolvent selected from primary or secondary alcohols or any mixture thereof,

[0030] d) evaporating the extract obtained in step c) to remove the co-solvent and obtain a crude extract in the form of a paste.

[0031] The invention secondly relates to a crude extract of spent sandalwood obtainable by the process according to the invention, characterised in that it comprises on the one hand 10 to 70%, preferably 12 to 23%, more preferably 14 to 21%, even more preferably 16 to 19% of volatile compounds and on the other hand 30 to 90%, preferably 77 to 88%, more preferably 79 to 86%, even more preferably 81 to 84% of semi-volatile to non-volatile compounds.

[0032] The present invention also relates to a solubilized extract of spent sandalwood obtainable by the process of the present invention, comprising 0.5 to 1.5% of a crude extract solubilized in a solvent selected from: a saturated or unsaturated, linear or branched fatty alcohol type solvent or a glyceride type solvent containing 8 to 30 carbon atoms or any mixture thereof.

[0033] The present invention thirdly relates to a cosmetic composition comprising: as active agent, a solubilized extract of spent sandalwood obtained according to the process of the invention, in a concentration ranging from 0.001 to 1%, and a physiologically acceptable medium.

[0034] The present invention fourthly relates to a cosmetic use of a composition comprising an extract of consumed sandalwood according to the invention for the care of the skin, scalp and appendages. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] The accompanying drawings illustrate by way of example the advantages and non-limiting embodiments resulting from the invention given in the description:

[0036] [ Figure 1 ] Analytical comparison of the DEDL chromatograms of the sandalwood essential oil described in Example 3 and the crude sandalwood extract according to Example 1.

[0037] [ Figure 2 ] Analytical comparison of the UV chromatograms at 300 nm of the sandalwood essential oil described in Example 3 and the crude sandalwood extract according to Example 1.

[0038] [ Figure 3 ] Quantification of OR2AT4 immunostaining in ex vivo skin biopsies. DETAILED DESCRIPTION

[0039] definition

[0040] Unless otherwise specified, all terms used in this specification have the most widely known meanings. For the purpose of the present invention, the following terms are defined as follows:

[0041] The term "spent sandalwood" is defined as sandalwood (Santalum album) trunks, twigs and chips that are recovered after extraction of the essential oil by steam distillation and then dried.

[0042] "Polar co-solvent" refers to a solvent having a polarity higher than that of CO2 in the supercritical state, such as a primary or secondary alcohol, or any mixture thereof.

[0043] "Glyceride solvents" refer to esters of fatty acids and glycerol.

[0044] When describing a range of values, the boundaries of the range should be understood to explicitly include the upper and lower limits of the range, as well as all intermediate values ​​within the range. For example, a range of values ​​from 1% to 10% should be understood to include 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% and 10%, as well as all decimal values ​​from 1% to 10%.

[0045] Unless otherwise indicated, numerical percentage values ​​are weight percentages, ie, the weight of the compound relative to the total weight of the subject mixture.

[0046] The compositions described herein may "comprise," "consist of," or "consist essentially of" essential compounds or optional ingredients.

[0047] The expression "consisting essentially of" means that the composition or component may include additional ingredients, but only if the additional ingredients do not alter the basic or novel characteristics of the composition or use described in the present application.

[0048] The term "cosmetically compatible solvent" means a solvent suitable for contact with the outer layers of the skin, scalp or appendages, which is not toxic, irritating, causes excessive allergic or similar reactions, or intolerance reactions, and which is proportional to a reasonable benefit / risk ratio at the concentrations used.

[0049] The term "active agent" refers to a compound capable of acting on the skin's functions with the aim of maintaining its integrity or capable of restoring it in the case of disorders caused by external factors (ultraviolet rays, pollution, detergents, etc.) or during aging with the aim of improving its appearance.

[0050] Extraction method

[0051] The sandalwood extracts described in the prior art have a strong olfactory aroma, characteristic of this wood, and contain non-negligible amounts of high-risk, volatile compounds, such as allergens.

[0052] The inventors wanted to develop an extract that did not have these characteristics. To do this, they chose to use a different raw material than natural sandalwood (trunk, branches or roots).

[0053] To produce the extract of the invention, the inventors chose to use sandalwood (Santalum album) chips that were recovered and then dried after the extraction of the essential oil by distillation. Therefore, the extract obtained does not contain volatile compounds, since the sandalwood has been consumed by extracting the essential oil by steam distillation. Therefore, it has a much less intense olfactory aroma and the allergen content is greatly reduced. In addition, the use of this perfume byproduct limits the impact on the environment and biodiversity and allows better management of plant resources.

[0054] The extraction method of the present invention has been extensively developed to optimize the extraction parameters. In this case, the effects of extraction temperature, percentage of inert compound used to increase the diffusivity of the extraction solvent, percentage of wetted material and co-solvent flow rate have been studied.

[0055] The present invention therefore firstly relates to a method for obtaining a sandalwood (Santalum album) extract, the method comprising the following steps:

[0056] a) adding 30 to 50% water to the consumed sandalwood;

[0057] b) optionally, mixing the spent sandalwood obtained in step a) with 1 to 20% of an inert compound to increase the diffusion rate of the extraction solvent;

[0058] c) extracting using a supercritical fluid selected from carbon dioxide (CO2) and xenon in the presence of a polar cosolvent selected from primary or secondary alcohols or any mixture thereof;

[0059] d) evaporating the extract obtained in step c) to remove the co-solvent and thereby obtain a crude extract in the form of a paste.

[0060] To carry out step a), water is added to the spent sandalwood to facilitate the penetration of the solvent into the interstitial phase of the wood to extract the extractable compounds. This step consists in extemporaneously adding 30 to 50% of water to the spent sandalwood.

[0061] The chips are very dry and porous and readily absorb the water added at this stage, leaving behind moist raw material.

[0062] Advantageously, dried, peeled sandalwood (Santalum album) trunk chips are used.

[0063] Advantageously, the consumed sandalwood may be from Santalum album (Santalum alba) grown in Australia.

[0064] Step b) is not mandatory but can improve the extraction process. In this step, the moistened spent sandalwood obtained in step a) is mixed with 1 to 20% of an inert compound to improve the diffusion rate of the extraction solvent, thereby avoiding preferential paths within the plant material. The inert compound is selected from organic or inorganic compounds, and preferably the inert compound is powdered cellulose.

[0065] Preferably, the inert compound is used in a concentration of 5 to 15%.

[0066] In the course of this description, the presence of inert compounds is not taken into account in the calculation of mass ratios and yields. The values ​​given take into account only the plant raw material.

[0067] To implement step c), a supercritical fluid selected from carbon dioxide (CO2) and xenon can be used. However, carbon dioxide is preferred.

[0068] For this step, a polar co-solvent is also used to increase the polarity of supercritical CO2, thereby facilitating the extraction of polar compounds.

[0069] The co-solvent is selected from a supercritical solvent having a higher polarity than CO2, which is selected from a primary alcohol or a secondary alcohol or any mixture thereof.

[0070] Preferably, the co-solvent is ethanol in water at a concentration of 80 to 100% (v / v), preferably at a concentration of 90 to 100% (v / v), and even more preferably at a concentration of 96% (v / v).

[0071] The mass ratio of the supercritical fluid (when said fluid is carbon dioxide) to the amount of raw material used (consumed sandalwood) is from 10 to 50, advantageously from 20 to 40 and preferably from 25 to 35.

[0072] The mass ratio of the supercritical fluid (carbon dioxide) to the co-solvent is from 0.050 to 0.080, advantageously from 0.055 to 0.075, and preferably from 0.060 to 0.070.

[0073] In step c), the extraction temperature is between 35 and 85°C, advantageously between 45 and 75°C and preferably between 55 and 65°C.

[0074] The pressure in the extractor is from 90 to 1000 bar, preferably from 150 to 700 bar, and even more preferably from 250 to 400 bar.

[0075] To implement step c), the mixture obtained in step a) or step b) is advantageously placed in a stainless steel cartridge and the cartridge is introduced into a supercritical fluid extractor. The solvent used for the extraction is carbon dioxide.

[0076] In step d), the first crude extract of the spent sandalwood obtained in step c) is recovered and evaporated under vacuum to completely remove the co-solvent. The evaporation temperature is up to 65°C and the pressure is below 90 mbar to allow good evaporation of the co-solvent.

[0077] At the end of step d), the extraction rate is preferably between 0.5 and 2%.

[0078] At this stage, a crude extract of consumed sandalwood in the sense of the present invention is obtained, which is in the form of a paste and contains:

[0079] -10 to 70% volatile compounds;

[0080] - 30 to 90% of semi-volatile to non-volatile compounds; said semi-volatile to non-volatile compounds comprising 0.35 to 3.5% of syringaldehyde.

[0081] The extraction can then continue with step e), wherein, in step e), the crude extract obtained in step d) is solubilized in a linear or branched saturated or unsaturated fatty alcohol solvent or a glyceride solvent containing 8 to 30 carbon atoms or any mixture thereof, preferably to obtain a crude extract at a concentration of 0.5 to 1.5 weight % based on the total weight of the solubilized extract.

[0082] Preferably, the fatty alcohol or glyceride solvent is selected from octyldodecanol, 2-hexyldecanol, oleyl alcohol and saturated vegetable fatty acid triglycerides (miglyol) (triglyceride mixture), or any mixture thereof.

[0083] Even more preferably, the solvent is octyldodecanol.

[0084] Advantageously, the extraction can be continued by an optional step f) in which the extract obtained in step d) or e) is purified. This step can be carried out by any technique known to a person skilled in the art, in particular by chromatography or by molecular distillation. This step can, for example, make it possible to standardize the extract.

[0085] Extracts

[0086] The extract of the present invention differs from the sandalwood extract obtained by supercritical CO2 in the prior art in that the raw material used consists of a by-product of distillation of white sandalwood chips, so that the material does not contain volatile compounds. The above method makes it possible to obtain an extract rich in polar and non-polar compounds. This extract has a wide chemical diversity, since it contains both residual portions of volatile compounds that were not extracted during steam distillation and semi-volatile to non-volatile compounds of varying polarity.

[0087] The present invention secondly relates to a crude extract of spent sandalwood obtainable by the process according to the invention.

[0088] The present invention also relates to a crude extract of spent sandalwood directly obtained by the process according to the invention.

[0089] "Raw extract" of spent sandalwood refers to the paste extract obtained in step d) of the process.

[0090] The crude extract of sandalwood consumed by itself contains:

[0091] - 10 to 70%, preferably 12 to 23%, more preferably 14 to 21%, even more preferably 16 to 19% of volatile compounds (mainly sesquiterpene alcohols, mainly (Z)-α-santalol, (Z)-β-santalol and derivatives of these compounds);

[0092] - 30% to 90%, preferably 77 to 88%, more preferably 79 to 86%, even more preferably 81 to 84% of semi-volatile to non-volatile compounds. This fraction consists mainly of lipid compounds represented by a mixture of acetylenic acids, fatty acids, and sesquiterpene derivatives of sandalwood (including dimers and aliphatic esters). The presence of smaller amounts of phenolic acids, aldehydes and lignans is also observed.

[0093] In a very advantageous embodiment, the crude extract comprises 16% of volatile compounds and 84% of semi-volatile to non-volatile compounds including 0.6% of syringaldehyde.

[0094] The term "volatile compounds" refers to organic molecules that can easily enter the gas phase at atmospheric pressure and room temperature. These substances are compounds with very low boiling points. They evaporate or sublime easily from their solid or liquid form. Gas chromatography is therefore the preferred technique for analyzing and detecting them in extracts. They are mainly sesquiterpene alcohols, mainly (Z)-α-santalenol and (Z)-β-santalenol.

[0095] The term "semivolatile to nonvolatile compounds" refers to molecules that do not readily enter the gas phase at atmospheric pressure and room temperature due to their higher boiling points than the volatile molecules. In the case of the extracts considered, these substances are medium polar molecules belonging to the family of phenolic compounds (phenolic acids and aldehydes, lignans), and nonpolar molecules belonging to chemical families such as fatty acids, acetylenic acids, sesquiterpenes obtained from sandalwood, and saponifiable compounds detected in the extracts.

[0096] Volatile compounds are analyzed by gas chromatography (GC) coupled to a mass spectrometer (MS) and / or a flame ionization detector (FID). Identifications are confirmed by comparing linear retention indices with mass spectra contained in a library. GC / FID quantification is accomplished by internal calibration using predicted and calculated response factors.

[0097] Semi-volatile to non-volatile compounds were monitored by high performance liquid chromatography (HPLC) coupled with a diode array UV detector (DAD) and a light scattering evaporation detector (LSED). The structural identification results were confirmed by nuclear magnetic resonance (NMR) examination and high resolution mass spectrometry (HRMS) analysis. If the standard is commercially available, the identification results can also be confirmed by injection of the standard. The total content of semi-volatile to non-volatile compounds was estimated by deducting the content of volatile compounds. Syringaldehyde constituted the majority of the medium polar compounds, which was considered as a marker for the crude extract and was quantified by external calibration with a diode array detector.

[0098] The main molecules identified were:

[0099] - In the volatile part, compounds of the sesquiterpene alcohol type, mainly (Z)-α-santalenol and (Z)-β-santalenol.

[0100] - In the semi-volatile to non-volatile fraction, fatty acids, acetylenic acids, saponifiable compounds, sesquiterpene derivatives of sandalwood as well as aldehydes and phenolic acids and lignans.

[0101] A non-exhaustive list of compounds present in these two fractions is given in Table 1 below:

[0102] [Table 1]

[0103]

[0104] Some markers of the crude extract can be quantified. They include, among others:

[0105] - 0.35 to 3.5%, advantageously 0.4 to 3%, preferably 0.45 to 2.5% of syringaldehyde;

[0106] - 1.3 to 2.3%, advantageously 1.4 to 2.2%, preferably 1.6 to 2.0% of (Z)-α-santalol;

[0107] - 0.7 to 1.3%, advantageously 0.8 to 1.2%, preferably 0.9 to 1.1% of (Z)-β-santalenol.

[0108] The present invention also relates to a solubilized extract of spent sandalwood obtainable by the process according to the present invention.

[0109] The present invention also relates to a solubilized extract of spent sandalwood directly obtained by the process according to the invention.

[0110] The term "extract of spent sandalwood" or "solubilized extract of spent sandalwood" in the sense of the present invention refers to the liquid extract obtained after solubilization in step e) of the process.

[0111] The solvent used in the solubilization step is selected from saturated or unsaturated, linear or branched fatty alcohol solvents containing 8 to 30 carbon atoms, or glyceride solvents, or any mixture thereof.

[0112] Preferably, the fatty alcohol or glyceride solvent is selected from octyldodecanol, 2-hexyldecanol, oleyl alcohol and saturated vegetable fatty acid triglycerides (triglyceride mixtures), or any mixtures thereof.

[0113] Even more preferably, the solvent is octyldodecanol.

[0114] The solubilized extract of spent sandalwood advantageously consists of 0.5 to 1.5% by weight of crude extract relative to the total weight of the extract solubilized in octyldodecanol.

[0115] Preferably, the consumed extract of sandalwood consists of 1.0% by weight of crude extract relative to the total weight of the extract solubilized in octyldodecanol.

[0116] The solubilized extract of the consumed sandalwood is in liquid form and contains a mixture of molecules having a wide range of polarities. Chromatographic analysis of the solubilized extract showed that the syringaldehyde content was 0.0035% to 0.035%.

[0117] Cosmetic composition

[0118] The invention thirdly relates to a cosmetic composition comprising, as active agent, an extract of consumed sandalwood according to the invention and a physiologically acceptable medium.

[0119] "Physiologically acceptable medium" means an excipient suitable for contact with the outer layers of the skin, scalp or appendages, which is non-toxic, irritating, unduly allergic or similar reactions or intolerance reactions and is proportional to a reasonable benefit / risk ratio.

[0120] Examples of physiologically acceptable media commonly used in the target application areas are formulation auxiliaries, such as solvents, thickeners, diluents, antioxidants, colorants, sunscreens, self-tanning agents, pigments, fillers, preservatives, fragrances, odor absorbers, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0121] Preferably, the composition according to the invention comprises an extract of consumed sandalwood obtainable by the process according to the invention in a concentration ranging from 0.001 to 1% by weight, preferably from 0.1% to 1%, relative to the total weight of the composition, and a physiologically acceptable medium.

[0122] The compositions that can be used according to the invention can be administered by any suitable route, in particular by external topical administration, and the formulation of the compositions will be adapted by those skilled in the art.

[0123] Preferably, the composition according to the invention is in a form suitable for topical administration.

[0124] The expression "topical application" refers to applying or spreading the composition comprising the extract of consumed sandalwood of the present invention on the surface of the skin, scalp, mucous membranes or appendages.

[0125] The term "skin" refers to the skin of the face (including the eyes and mouth, nose, forehead), neck, hands, and also the skin of the entire body (including the scalp).

[0126] The term "scalp" refers to the skin covering the skull, including the hair follicles and the interfollicular skin spaces.

[0127] The term "appendages" refers to the keratinized skin appendages present in humans and animals, which are rich in keratin, more particularly the hair, eyelashes, eyebrows and nails.

[0128] The compositions according to the invention are particularly suitable for topical application to healthy skin.

[0129] For the purposes of the present invention, healthy skin is defined as skin that is free of skin pathology.

[0130] The topical compositions used to implement the invention may especially be in the form of aqueous, aqueous-alcoholic or oily solutions, oil-in-water emulsions, water-in-oil emulsions, multiple emulsions, microemulsions, nanoemulsions or any colloidal system usable in cosmetics; they may also be in the form of suspensions or powders suitable for application to the skin, mucous membranes, lips and / or hair.

[0131] These compositions can be more or less fluid and can also be in the form of a cream, lotion, milk, serum, ointment, gel, paste or foam. They can also be in solid form, such as a stick, or be formulated to be compatible with aerosol delivery.

[0132] In all cases, the skilled person will ensure that the adjuvants and their proportions are chosen in a way that does not impair the advantageous properties sought for the composition according to the invention. These adjuvants may, for example, correspond to 0.01 to 20% of the total weight of the composition. When the composition according to the invention is an emulsion, the fatty phase may be 5 to 80% by weight, preferably 5 to 50% by weight, relative to the total weight of the composition. The emulsifiers and co-emulsifiers used in the composition are selected from those conventionally used in the field under consideration. For example, they may be used in a proportion of 0.3 to 30% by weight relative to the total weight of the composition.

[0133] In particular embodiments, the compositions may contain one or more additional active agents to enhance the effects of the sandalwood extract used in the present invention.

[0134] The INCI Dictionary & Handbook ("International Nomenclature of Cosmetic Ingredients 13th Edition 2010" published by the Personal Care Products Council, Inc., Washington, D.C.) describes a wide variety of non-limiting cosmetic ingredients commonly used in the skin care industry that are suitable for use as additional active agents in the compositions according to the present invention.

[0135] Non-limiting examples of these categories of additional active agents include: anti-aging agents, anti-wrinkle agents, moisturizers, softeners, keratolytic agents, or desquamating agents. agents, anti-seborrheic agents, anti-dandruff agents, agents that regulate skin cell differentiation or proliferation, agents that regulate skin pigmentation, self-tanning agents, anti-atmospheric pollution agents, anti-glycation agents, firming agents, aquaporin synthesis stimulators, lipid and stratum corneum component (ceramide, fatty acid) synthesis stimulators, adipocyte proliferation stimulators, glycosaminoglycan synthesis stimulators, DNA repair agents, DNA protective agents, sensitive skin treatment and / or care agents, firming agents, anti-stretch mark agents, astringents, dermo-relaxing agents, cytokine growth factors, agents acting on capillary circulation and / or microcirculation, agents inhibiting vascular permeability, agents acting on cell metabolism, agents improving the dermal-epidermal junction, agents inducing head hair and / or other hair growth, lipolysis stimulators, slimming agents, anti-cellulite agents agents), sunscreens, agents capable of reducing or treating eye bags, and mixtures thereof, as long as they are physically and chemically compatible with the other ingredients of the composition, especially with the active ingredients of the present invention.

[0136] Furthermore, the nature of these additional active agents must not alter the benefits of the active ingredients of the invention in an unacceptable way.These additional active agents may be synthetic or natural, such as plant extracts, or may be derived from a biological fermentation process.

[0137] Such additional active agents can also be selected according to their chemical composition from the following: amino sugars, glucosamine, D-glucosamine, N-acetylglucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetylmannosamine, galactosamine, N-acetylgalactosamine, vitamin B3 and its derivatives, nicotinamide, sodium dehydroacetate, dehydroacetic acid and its salts, phytosterols, salicylic acid compounds, hexamidine, dialkanoyl dihydroxyproline compounds, soybean extracts and derivatives, equol, isoflavones, flavonoids, phytosamine. triol (phytantriol), farnesol, geraniol, bisabolol, peptides and their derivatives, dipeptides, tripeptides, tetrapeptides, pentapeptides and hexapeptides and their derivatives, lysine-threonine-threonine-lysine-serine (lys-thr-thr-lys-ser), palmitoyl-lysine-thr-lysine-serine (palmitoyl-lys-thr-lys-ser), carnosine, N-acyl amino acid compounds, retinoids (retinoid ), retinyl propionate, retinol, retinyl palmitate, retinyl acetate, retinal, retinoic acid, water-soluble vitamins, ascorbic acid esters, vitamin C, ascorbyl glucoside, ascorbyl palmitate, magnesium ascorbyl phosphate, sodium ascorbyl phosphate, vitamins and their salts and derivatives, provitamins and their salts and derivatives, ethyl panthenol, vitamin A and its derivatives, vitamin B and its derivatives, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin F, vitamin Vitamin K and its derivatives, pantothenic acid pantothenyl ether, panthenol and its derivatives, ethyl panthenol, d-panthenol, biotin, amino acids and their salts and derivatives, water-soluble amino acids, asparagine, alanine, indole, glutamic acid, water-insoluble vitamins, beta-ionol, cedarwood alcohol, and their derivatives, water-insoluble amino acids, tyrosine, tryptamine, granular materials, butylated hydroxytoluene, butylated hydroxyanisole, allantoin, tocopherol nicotinate nicotinate), tocopherol, tocopherol ester, palmitoyl-gly-his-lysine, phytosterol, hydroxy acid, glycolic acid, lactic acid, lactobionic acid, keto acid, pyruvic acid, phytic acid, lysophosphatidic acid, stilbene, cinnamate, resveratrol, kinetinzeatin, dimethylaminoethanol, natural peptide, soy peptide, acid sugar salt, manganese gluconate, zinc gluconate, piroctone olamine, 3,4,4'-Trichlorocarbanilide, triclocarban, zinc pyrithione, hydroquinone, kojic acid ascorbyl, magnesium ascorbyl phosphate, ascorbyl glucoside, pyridoxine, aloe, terpene alcohols, allantoin, bisabolol, dipotassium glycyrrhizinate, glyceric acid, sorbitol, pentaerythritol, pyrrolidone and its salts, dihydroxyacetone, erythrulose, glyceraldehyde, tartaraldehyde, clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, witch hazel distillate, copolymer of eicosene and vinyl pyrrolidone, iodopropyl butylcarbamate, polysaccharides, essential fatty acids, salicylates, glycyrrhetinic acid, carotenoids, ceramides and pseudoceramides, complex lipids, oils of substantially natural origin (e.g., avocado oil, apricot oil, evening primrose oil, prune oil) oil), palm oil, monooiil, kahai oil), hydroquinone, HEPES, procysteine, O-octanoyl-6-D-maltose, disodium salt of methylglycine diacetic acid, steroids (e.g. dioscin and DHEA derivatives, DHEA dehydroepiandrosterone and / or chemical or biological precursors or derivatives), N-ethylcarbonyl-4-p-aminophenol, alpha-hydroxy acids, beta-hydroxy acids, moisturizers, epidermal hydrolases, plant extracts, plant hormones, yeast extracts, metalloproteinase inhibitors, enzymes, enzyme inhibitors, enzyme inducers, coenzymes, chelating agents, plant extracts and plant derivatives, essential oils, marine extracts, agents from biofermentation and / or biotechnological processes, mineral salts, cell extracts.

[0138] The following substances may be mentioned as further examples:

[0139] - Peptides known commercially as: CHRONOGEN TM LAMINIXYL IS TM PEPTIDE Q10 TM 、COLLAXYL TM (Patent FR2827170, )、PEPTIDEVINCI01 TM (Patent FR2837098, )、PEPTIDE VINCI02 TM (Patent FR2841781, ), ATPeptide TM (Patent FR2846883, ); or a synthetic peptide of sequence Arg-Gly-Ser-NH2, which is named ATPeptide TM Depend on Sale;

[0140] - Artemia salina extract, which is known under the name GP4G TM (FR2817748, )Sale;

[0141] - Plant peptide extracts, such as flax extract (Lipigenin TM , Patent FR2956818, ), extracts of soy, spelt, vine, rapeseed, flax, rice, corn, pea, and cocoa;

[0142] -Yeast extract, such as Dynagen TM (Patent FR2951946, ) or Actopontine TM (Patent FR2944526, );

[0143] use

[0144] The present invention also relates to a cosmetic use of a composition comprising an extract of consumed sandalwood according to the invention for the care of the skin, scalp and appendages.

[0145] The cosmetic use according to the invention relates to a method of cosmetic treatment by topical application to healthy skin.

[0146] The invention also relates to the cosmetic use of a composition according to the invention for improving the appearance of the skin, combating signs of skin aging, increasing the skin's ability to detect external pollutants and restoring the lipid barrier function of the skin.

[0147] The invention also relates to the cosmetic use of a composition according to the invention for increasing the expression of the olfactory receptor (OR2AT4) in the skin.

[0148] The invention also relates to the cosmetic use of a composition according to the invention for lightening the skin tone.

[0149] The invention also relates to the cosmetic use of a composition according to the invention for promoting hair growth.

[0150] The expression "detecting external pollutants" refers to the ability of the skin to trigger its defense mechanisms to reduce the effects of exposure to any type of atmospheric pollutants, including in particular odorous molecules of the volatile organic compound type.

[0151] The expression "improving the appearance of skin" means reducing damage caused by environmental stressors in order to restore skin function and limit visible damage to the skin, such as signs of aging or skin sensitivity.

[0152] The term "skin sensitivity" refers to the reaction of healthy skin to various insults with signs of discomfort (such as tingling, warmth or tightness of the skin of the face or scalp) and possible visible reactions (such as redness).

[0153] The term "signs of skin aging" refers to changes in the appearance of the skin that occur due to aging, selected from wrinkles and fine lines, creases, eye bags, dark circles, withering, loss of skin elasticity, firmness and / or tone, irregularities in skin texture or skin tone; and all internal changes in the skin that do not systematically lead to changes in appearance, such as thinning of the skin, or all internal degeneration of the skin after environmental stress (or external aggression).

[0154] The expression "improving lipid barrier function or skin barrier function" means that the protective performance of the skin against external aggressions is improved to maintain the integrity of the skin.

[0155] The consumed sandalwood extracts of the present invention were tested for olfactory receptor (OR2AT4) expression in the skin.

[0156] Odor perception involves olfactory (or smell) receptors on the nasal epithelium. Activation of these receptors by odor molecules results in a specific sensation, which can act as an alarm or communication signal. Olfactory receptors belong to the family of G protein-coupled receptors. They are activated by odor molecules, inducing calcium signals, which indicates their function. In addition to the nasal epithelium, ectopic expression of certain types of olfactory receptors has been reported in different organs, including the skin (Denda M. Newly discovered olfactory receptors in epidermal keratinocytes are associated with proliferation, migration, and re-epithelialisation of keratinocytes. J Invest Dermatol. 2014 Nov; 134 (11): 2677-2679). In fact, during evolution, some species have retained olfactory receptors expressed on the surface of their skin. Generally, they are involved in the recognition of environmental signals. Among these receptors, the role of OR2AT4 type receptors in human skin has been studied in more detail. These receptors have the synthetic molecule sandalwood 210 (Sandalore) as their ligand and play a role in skin regeneration processes by interfering with the proliferation and migration of keratinocytes (Busse D. et al., A synthetic sandalwood odorant induces wound-healing processes in human keratinocytes via the olfactory receptor OR2AT4. J Invest Dermatol. 2014 Nov; 134 (11): 2823-2832). The expression of OR2AT4-like receptors in hair follicles has also been reported, where they are associated with the hair growth phase (Chéret J. et al., Olfactory receptor OR2AT4regulates human hair growth. Nat Commun 9, 3624, 2018). Other receptor types have been described in the skin, including keratinocytes and melanocytes (Wojcik S, Weidinger D, S, Luger T, Hatt H, Jovancevic N. Functional characterization of the extranasal OR2A4 / 7expressed in human melanocytes. Exp Dermatol. 2018;27(11):1216-1223).

[0157] Example

[0158] The present invention is illustrated by the following non-limiting examples:

[0159] Example 1 : Preparation of an extract of consumed sandalwood

[0160] For the purposes of the examples, the Sandalwood (Santalum album) tree is grown in Australia.

[0161] As white sandalwood is not native to Australia and the raw material comes from private plantations on non-Commonwealth landholdings, no permit is required under Part 8A of the EPBC Act (Environment Protection and Biodiversity Conservation Act 1999), which governs, inter alia, access to genetic resources and benefit sharing (ABS).

[0162] The plant parts used consisted of spent bark-free chips (i.e. recovered after steam distillation) of the trunk of the white sandalwood tree. During the development of the method, several parameters were studied, namely the wettability of the plant material, the percentage of added cellulose, the extraction temperature and the co-solvent flow rate.

[0163] Table 2 shows some simulation results obtained from the experimental design performed to optimize the extraction parameters:

[0164] [Table 2]

[0165]

[0166]

[0167] The extraction conditions are selected to obtain a satisfactory extraction yield while ensuring a beneficial chemical diversity of the extract, in particular a correct content of phenolic compounds. The preferred extraction parameters are as follows:

[0168] The cuttings are moistened with 40% (wt%) water, mixed with 10% powdered cellulose and then placed in a stainless steel cartridge. This cartridge is introduced into a supercritical fluid extractor, such as a Separex SFE 5 extractor. The extraction solvent used is supercritical carbon dioxide at a flow rate of 15 Kg / h, supplemented with 96% (volume / volume) ethanol in water as a polar cosolvent at a flow rate of 20 ml / min. The mass ratio of carbon dioxide to the moistened plant is 30, and the mass ratio of carbon dioxide to cosolvent is 0.065. The pressure and temperature in the extractor are 300 bar and 60° C., respectively. The pressure and temperature in the separator are 55 bar and 35° C., respectively. The extract thus obtained is evaporated under vacuum until the ethanol is completely evaporated (pressure is less than 90 mbar at a water bath temperature of 65° C.). The extraction yield is 1.2%.

[0169] The crude extract obtained was in the form of a paste.

[0170] The crude extract was solubilized in agro-sourced octyldodecanol to obtain a solubilized extract which appeared to be a clear fluid solution containing 1.0% of the crude extract of Santalum Albizia Sinensis chips.

[0171] Example 2 : Characterization of the crude extract of consumed sandalwood obtained according to Example 1

[0172] A non-exhaustive list of the compounds present in each fraction is detailed in Table 3 below:

[0173] [Table 3]

[0174]

[0175]

[0176] Some markers of the crude extract can be quantified. The extract contains in particular:

[0177] - 0.6% syringaldehyde;

[0178] -1.8% (Z)-α-santalol;

[0179] -1.0% (Z)-β-santalol;

[0180] -16% volatile fraction;

[0181] -84% semi-volatile to non-volatile fraction.

[0182] Embodiment 3: Preparation of sandalwood essential oil extract

[0183] For comparative analysis, the essential oil extraction of (unconsumed) sandalwood was carried out in a conventional manner by steam distillation. The bark-free trunks, branches and roots of white sandalwood are dried and then coarsely ground into chips, which are placed in a separate distiller and then passed through a stream of steam; this steam releases volatile molecules or essential oils, which are carried away by the steam and condensed in a condenser. Since the essential oils have a lower density than water and are not water-soluble or only slightly soluble in water, the essential oils are collected at the outlet in an oil-water separator called an essencier. The water that still contains traces of essential oils is called hydrolate.

[0184] The essential oil is obtained by making a communal mixture from the three oils made previously. It is a pale yellow to yellow liquid.

[0185] Embodiment 4: Identification of the main phytochemical differences between the essential oil and the extract obtained in Example 1

[0186] The crude extract of spent sandalwood obtained in Example 1 and the essential oil of white sandalwood prepared in Example 3 were compared by subjecting solutions of these samples at the same concentration to high performance liquid chromatography (HPLC) coupled with a diode array detector and a DEDL detector. Chromatography was performed on a core-shell C18 column with an elution gradient using an acidified mobile phase consisting of a water / CAN / IPA mixture for lane A and ACN / IPA / MeOH for lane B. Figure 1 and 2 DEDL and UV chromatograms at 300 nm of the sandalwood essential oil described in Example 3 and the crude extract of the depleted sandalwood according to Example 1 are shown, which were analyzed on a core-shell C18 column in HPLC / UV-DEDL with gradient elution (0 to 5 minutes 100% A, 5 to 22 minutes from 100% A to 100% B, then 22 to 32 minutes 100% B - A: H2O / ACN / IPA / HCO2H 95 / 2.5 / 2.5 / 0.1 (v / v / v / v) and B: IPA / ACN / MeOH / HCO2H 40 / 40 / 20 / 0.1 (v / v / v / v)). The y-axis shows the detector response in mV for the DEDL graph and in mAU for the diode array graph. In both cases, the abscissa represents the analysis time in minutes.

[0187] like Figure 1 and 2 As shown in , lipids and phenolic compounds (semi-volatile to non-volatile compounds) were not detected or were only weakly detected in sandalwood essential oil.

[0188] Embodiment 5: Evaluation of the Effect of Consumed Sandalwood Extract of Example 1 on OR2AT4 Olfactory Receptor Expression in Human Skin Biopsies

[0189] The aim of this study was to demonstrate the effect of sandalwood extract on the synthesis of the olfactory receptor OR2AT4 in cultured human skin biopsies.

[0190] plan:

[0191] The expression of OR2AT4 was assessed by indirect immunofluorescence on skin biopsies pretreated by topical application of 0.1%, 0.5% and 1% (volume / volume) of the solubilized extract of consumed sandalwood of Example 1 for 48 hours (twice a day). For this purpose, the solubilized extract of consumed sandalwood from Example 1 was diluted to 0.1%, 0.5% and 1% (volume / volume) with octyldodecanol. Similar dilutions of sandalwood essential oil from Example 3 or the synthetic analogue Santalum 210 were provided to other biopsies. Control biopsies incubated in parallel under the same conditions received a placebo (octyldodecanol). After incubation, the biopsies were fixed and embedded in paraffin for tissue sectioning. Detection of the OR2AT4 receptor was performed by incubation with an anti-OR2AT4 antibody (Novus). After one and a half hours of incubation, the sections were rinsed and then the sections were incubated with a fluorophore (Alexa Fluor). The sections were then incubated in the presence of an anti-rabbit secondary antibody coupled to 488 (Invitrogen). The sections were then examined under an epifluorescence microscope (Zeiss Axiovert 200M microscope). The expression of OR2AT4 was then observed and analyzed by image analysis ( Image analysis software (Improvision) was used for quantification.

[0192] result:

[0193] like Figure 3 As shown in Figure 1, when biopsies were treated with 0.1%, 0.5% and 1% sandalwood extract, OR2AT4 expression increased by 95%, 81% and 73%, respectively. Sandalwood essential oil tested under the same conditions did not cause any increase in OR2AT4 expression, and even induced a decrease at 0.5% and 1%. For Santalum 210, a known ligand for this receptor, less than 1% application did not seem to have any effect on receptor expression. However, after applying 1%, 3% and 5% sandalwood 210, OR2AT4 expression increased by 28%, 72% and 71%, respectively.

[0194] in conclusion:

[0195] Sandalwood extract showed a positive effect on the expression of olfactory receptor OR2AT4.

[0196] Embodiment 6: Evaluation of the effect of the consumed sandalwood extract of Example 1 on the homeostasis of the epidermal barrier by studying filaggrin:

[0197] The aim of this study was to demonstrate the effect of sandalwood extract on epidermal barrier homeostasis by studying filaggrin, a protein involved in the maintenance of cohesion and hydration of the upper epidermal layers, which is essential for maintaining the homeostasis of the skin barrier.

[0198] plan:

[0199] The expression of filaggrin was assessed by indirect immunofluorescence on skin biopsies pretreated by topical application of a solubilized extract of consumed sandalwood from Example 1 diluted to 0.1% (vol / vol) with octyldodecanol for 48 hours (twice a day). In parallel, the biopsies were treated with sandalwood essential oil from Example 3 from the same geographical location and tested under the same conditions. A placebo (octyldodecanol) was administered to control biopsies incubated in parallel under the same conditions. After incubation, the biopsies were fixed and embedded in paraffin for tissue sectioning. Detection of filaggrin was performed by incubation with an anti-filaggrin antibody (Santa Cruz). After one and a half hours of incubation, the sections were rinsed and then the sections were incubated with a fluorophore (Alexa Fluor). The sections were then incubated in the presence of an anti-mouse secondary antibody coupled to 488 (Invitrogen). The sections were then examined under an epifluorescence microscope (Zeiss Axiovert 200M microscope). The expression of filaggrin was then observed and analyzed by image analysis ( Image analysis software (Improvision) was used for quantification.

[0200] result:

[0201] When the biopsy tissue was treated with 0.1% sandalwood extract, the expression of filaggrin increased by 28%. Sandalwood essential oil neither caused any increase nor a decrease in filaggrin expression. In parallel, the biopsy tissue treated with EGCG (epigallocatechin gallate), a positive control for filaggrin, showed an increase of +27% in filaggrin.

[0202] in conclusion:

[0203] Sandalwood extract showed a positive effect on filaggrin expression, suggesting an effect on enhancing skin barrier function.

[0204] Embodiment 7: Evaluation of the effect of the consumed sandalwood extract of Example 1 on lipid barrier function disrupted by air pollution stress by studying ceramide synthase 3:

[0205] The aim of this experiment was to demonstrate the effect of sandalwood extract on the homeostasis of the epidermal barrier disrupted by pollution stress, by studying ceramide synthase 3 in cultured human skin biopsies. Ceramide synthase 3 allows the formation of ceramides from sphingosine. Ceramides are members of the sphingolipid family that allow the establishment of barrier functions but are also bioactive metabolites involved in epidermal renewal. The pressure applied in this experiment was achieved with ultrafine particles from a diesel engine.

[0206] plan:

[0207] The expression of the enzyme ceramide synthase 3 was assessed by indirect immunofluorescence on skin biopsies, which were stressed by applying ultrafine particles from diesel emission at 500 μg / ml for 16 hours and then treated by topical application of the solubilized extract of consumed sandalwood from Example 1 further diluted with octyldodecanol to 0.1%, 0.5% and 1% (volume / volume percentage) for 48 hours (twice a day). Control biopsies incubated in parallel under the same conditions received a placebo (octyldodecanol). At the end of the incubation, the biopsies were fixed and embedded in paraffin for tissue sections. The detection of the enzyme ceramide synthase 3 was performed by incubation with an anti-ceramide synthase 3 antibody (Novus). After one and a half hours of incubation, the sections were rinsed and then the sections were incubated with a fluorophore (Alexa Fluor). The sections were then incubated in the presence of an anti-rabbit secondary antibody coupled to 488, Invitrogen). The sections were then examined under an epifluorescence microscope (Zeiss Axiovert 200M microscope). The expression of ceramide synthase 3 was then observed and analyzed by image analysis ( Image analysis software (Improvision) was used for quantification.

[0208] result:

[0209] After applying ultrafine diesel particles, the expression of ceramide synthase 3 decreased by -39%. When the biopsies were stressed with ultrafine diesel particles and additionally treated with 0.1% and 0.5% sandalwood extract, the expression of ceramide synthase 3 decreased by only -29% and -18%, respectively. After applying 1% sandalwood extract, the expression level of ceramide synthase 3 even returned to the basal state without stress, indicating that sandalwood has a positive effect on this lipid component of the skin barrier.

[0210] in conclusion:

[0211] Ultrafine particle pollution in the atmosphere negatively affects the expression of ceramide synthase 3. The reduction in the expression of this enzyme was reduced after the application of sandalwood extract. Sandalwood extract helps to strengthen the lipid barrier function in the presence of this type of pollutant stress.

[0212] Embodiment 8: Evaluation of the lightening potential of the consumed sandalwood extract of Example 1 on ex vivo skin biopsies

[0213] principle:

[0214] The aim of this study was to evaluate the lightening potential of sandalwood extract on ex vivo skin biopsies using Fontana-Masson histological melanin staining, based on the reduction of an ammoniacal silver nitrate solution to metallic silver. The obtained staining revealed the melanin content, which was quantified by image analysis.

[0215] plan:

[0216] Ex vivo human skin biopsies were cultured and treated for 48 hours with the solubilized extract of the consumed sandalwood of Example 1 or the essential oil of sandalwood of Example 3 diluted to 0.1% and 1% (vol / vol) in octyldodecanol. After treatment, the biopsies were fixed for histological analysis and embedded in paraffin. After dewaxing, the sections were incubated with an ammoniacal silver nitrate solution at 60° C. for 10 minutes. After rinsing, they were treated with 5% sodium thiosulfate for 2 minutes, rinsed again, and mounted for examination under an Eclipse E600 microscope (Nikon). The photographs were taken with a QImaging Retiga2000R Fast1394 camera and analyzed with Q-Capture Pro 7 software (QImaging).

[0217] result:

[0218] On ex vivo skin biopsies, a decrease of -50% and -46% in melanin content was observed following application of sandalwood extract at 0.1% and 1%, respectively (highly significant by Student's t-test compared to placebo biopsies), while sandalwood essential oil showed smaller decreases of -25% and -21% at 0.1% and 1%, respectively.

[0219] in conclusion:

[0220] This test therefore concluded that sandalwood extract has a potential lightening effect on ex vivo skin biopsies.

[0221] Embodiment 9: Evaluation of the effect of the consumed sandalwood extract of Example 1 on the activity of the dermal papilla cells of the hair follicles:

[0222] principle:

[0223] Stimulation of the OR2AT4 olfactory receptor in the hair follicle is associated with the hair growth phase, known as the anagen phase. A marker for this phase is the expression of IGF-1 (insulin-like growth factor 1) in dermal papilla cells. In this example, human dermal papilla cells were treated with the consumed sandalwood extract of Example 1 and then analyzed for the expression of IGF-1 by immunostaining. An increase in this marker indicates that the hair follicles are still in the anagen phase.

[0224] plan:

[0225] Human dermal papilla cells (HDPC) were cultured in the presence of the solubilized extract of consumed sandalwood from Example 1 diluted to 0.001 and 0.005% with DMSO. Control cells received an equivalent dilution of DMSO. Treatment was performed once a day for 48 hours. The cells were then fixed on their supports for immunolabeling. They were incubated overnight in the presence of an anti-IGF-1 antibody (mouse monoclonal, SantaCruz). The cells were then rinsed and incubated for 1 hour with a secondary anti-mouse antibody coupled to the fluorescent marker AlexaFluor 488 (Invitrogen). After another washing step, the cells were observed under an epifluorescence microscope (Zeiss Axiovert 200M microscope) and analyzed by image analysis ( The fluorescence intensity was quantified using image analysis software, Improvision.

[0226] Results and Conclusions:

[0227] Human dermal papilla cells showed an increase of +27% and +10% in IGF-1 expression at 0.001% and 0.005% concentrations of sandalwood extract, respectively. This result indicates that sandalwood extract is active in maintaining the hair growth phase.

[0228] Embodiment 10: Formulation of cream for topical application

[0229] [Table 4]

[0230]

[0231]

Claims

1. A method for obtaining a crude sandalwood extract, the method comprising the following steps: a) adding 30 to 50% water to spent sandalwood, wherein the spent sandalwood is sandalwood trunks, branches and root chips recovered and dried after extracting essential oil by steam distillation; b) mixing the moistened spent sandalwood with 1 to 20% of an inert compound; c) extracting using a supercritical carbon dioxide fluid in the presence of 80 to 100% ethanol as a polar co-solvent in a concentration of 80 to 100% by volume / water volume percentage, wherein the weight ratio of the supercritical carbon dioxide fluid to the spent sandalwood used is 10 to 5, the extraction temperature is 35 to 85°C, and the pressure in the extractor is 9 0 to 1000 bar, the weight ratio of the supercritical carbon dioxide fluid to the polar co-solvent is 0.050 to 0.080; d) evaporating the extract obtained in step c) to remove the polar co-solvent and thereby recovering a crude extract in a paste state, wherein the crude extract obtained comprises 10 to 70% of volatile compounds and 30 to 90% of semi-volatile to non-volatile compounds, and the semi-volatile to non-volatile part comprises 0.45 to 2.5% of syringaldehyde, and the volatile part comprises 1.6 to 2.0% of (Z)-α-santalol and 0.9 to 1.1% of (Z)-β-santalol.

2. The process according to claim 1, wherein in step b) the moistened spent sandalwood is optionally mixed with 5 to 15% of an inert compound.

3. The method according to claim 1, wherein in step c) the polar co-solvent is ethanol having a concentration of 90 to 100% in terms of volume / water volume percentage.

4. The method according to claim 1, wherein in step c), the polar co-solvent is ethanol having a concentration of 96% by volume / water volume percentage.

5. The method according to claim 1, wherein in step c) the weight ratio of the supercritical carbon dioxide fluid to the polar co-solvent is 0.055 to 0.

075.

6. The method according to claim 1, wherein in step c) the weight ratio of the carbon dioxide fluid in a supercritical state to the polar co-solvent is 0.060 to 0.

070.

7. The method according to claim 1, wherein in step c), the extraction temperature is 45 to 75°C.

8. The method of claim 1, wherein in step c), the extraction temperature is 55 to 65°C.

9. The process according to claim 1, wherein in step c), the pressure in the extractor is 150 to 700 bar.

10. The process according to claim 1, wherein in step c), the pressure in the extractor is 250 to 400 bar.

11. The process according to claim 1, wherein the crude extract obtained in step d) is dissolved in a saturated or unsaturated, linear or branched fatty alcohol solvent or a glyceride solvent containing 8 to 30 carbon atoms or any mixture thereof to obtain 0.5 to 1.5 wt% of a soluble extract relative to the total weight of the crude extract.

12. The process according to claim 11, wherein the crude extract obtained in step d) is dissolved in a solvent selected from octyldodecanol, 2-hexyldecanol, oleyl alcohol and a mixture of triglycerides to obtain 0.5 to 1.5 wt% of soluble extract relative to the total weight of the crude extract.

13. A crude sandalwood extract obtainable by the process according to claim 1, characterized in that It contains: 10 to 70% volatile compounds, and 30 to 90% semi-volatile to non-volatile compounds.

14. The sandalwood crude extract according to claim 13, characterized in that It contains: 12 to 23% volatile compounds.

15. The sandalwood crude extract according to claim 13, characterized in that It contains: 14 to 21% volatile compounds.

16. The sandalwood crude extract according to claim 13, characterized in that It contains: 16 to 19% volatile compounds.

17. The sandalwood crude extract according to claim 13, characterized in that It contains: 77 to 88% semi-volatile to non-volatile compounds.

18. The sandalwood crude extract according to claim 13, characterized in that It contains: 79 to 86% semi-volatile to non-volatile compounds.

19. The sandalwood crude extract according to claim 13, characterized in that It contains: 81 to 84% semi-volatile to non-volatile compounds.

20. A soluble extract obtainable by the process according to claim 11, characterized in that It contains 0.0035% to 0.035% of syringaldehyde in semi-volatile to non-volatile compounds, and the solvent is octyldodecanol.

21. A composition comprising as active agent a soluble extract according to claim 20 in a concentration of 0.001 to 1% and a physiologically acceptable medium.

22. A composition comprising as active agent a soluble extract according to claim 20 in a concentration of 0.1 to 1% and a physiologically acceptable medium.

23. Use of the composition according to claim 21 in preparing cosmetics for caring for skin and appendages, wherein the use is selected from improving skin appearance, restoring the lipid barrier function of the skin or promoting hair growth, wherein the appendages are eyelashes, eyebrows and nails.

24. Use of the composition according to claim 21 in preparing cosmetics for caring for the scalp, wherein the use is selected from improving skin appearance, restoring the lipid barrier function of the skin or promoting hair growth.

25. The use according to claim 23, wherein the cosmetic is used to increase the expression of olfactory receptor OR2AT4.

26. The use according to claim 23, wherein the cosmetic is used to combat signs of skin aging or to brighten the skin tone.

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