Method for obtaining a rosewood extract, compositions comprising same and cosmetic uses thereof

Abstract

The invention relates to a rosewood extract obtained from co-products of the distillation of rosewood (Aniba rosaeodora) essential oil by extraction using supercritical CO2 in the presence of a co-solvent. The subject matter of the invention is also a method for obtaining such an extract. The invention further relates to the cosmetic compositions comprising such an extract and finally to the cosmetic uses of such compositions for firming the skin, limiting the appearance of ptosis, redefining the contour of the face, and limiting the appearance of wrinkles and fine lines.

Description

ISP‐BV23‐200PCT Description Description Title of the invention: PROCESS FOR OBTAINING A ROSEWOOD EXTRACT, COMPOSITIONS COMPRISING IT AND ITS COSMETIC USES Technical field

[0001] The present invention relates to the field of skin care ingredients. The invention relates to a rosewood extract (Aniba rosaeodora) composed inter alia of nicotinic acid, cotoine, sesquiterpene derivatives and depleted in volatile compounds and anibine, obtained by a supercritical CO2 extraction process from the co-products of rosewood distillation. The invention also relates to cosmetic compositions comprising such an extract, and finally to the uses of such compositions for preventing or limiting the signs of skin aging. Technical background of the invention

[0002] Rosewood (Aniba rosaeodora Ducke or "rosewood" in English) is a tree found in the tropical and subtropical rainforests of the Amazon basin (Venezuela, Colombia, Ecuador, French Guiana, Guyana, Peru, Suriname and Brazil).

[0003] The trunk, branches or leaves of rosewood are used in perfumery and cosmetics in the form of essential oil whose olfactory particularity is conferred by linalool, a volatile terpene alcohol present in large quantities in the essential oil (70 to 90%).

[0004] According to the literature, the non-volatile part of rosewood contains anibine, 4-methoxyparacotoine, cotoine and pinocembrine, but also (-)-rubranine, a chalcone present in the branches of this species (Mors, WB et al. J. Am. Chem. Soc., 1957, 79, 4507-4511; Gottlieb, OR^and Mors, WB, J. Am. Chem. Soc., 1958, 80, 2263-2265; Galaverna, RS et al., Anal. Methods, 2015, 7, 1984-1990; De Alleluia, IB^et al., Phytochem., 1978, 17, 517-521).

[0005] Several chemical classes of compounds present in rosewood, including aryl phenyl ketones, nicotinic acid, and phytosterols, have shown biological activities such as antioxidant, antibacterial, antimicrobial, antifungal, anti-inflammatory, anticancer, vasodilatory, and hypolipidemic properties (Wu, S.-B.^et al., Nat. Prod. Rep., 2014, 31, 1158-1174). ISP‐BV23‐200PCT Description

[0006] The rosewood extracts described in the prior art are mainly obtained by hydrodistillation. This results in extracts enriched in volatile compounds and mainly in linalool (70 to 90%). However, a significant quantity of these volatile compounds, including linalool and benzyl benzoate, are considered allergenic.

[0007] The inventors wanted to develop an extract that did not have these characteristics in terms of chemical composition and olfactory note. To do this, they chose to use a rosewood raw material (trunk, branches and leaves) that had undergone a first distillation and was therefore exhausted (or depleted) in volatile odorous compounds. This exhausted raw material is again extracted by a supercritical fluid in order to obtain an extract enriched in semi- to non-volatile compounds and possessing a great chemical diversity.

[0008] To date, to the inventors' knowledge, no document describes the parameters of a rosewood extraction by a supercritical fluid. For example, documents FR1000801B1, CN105708740 and CN105640822 describe cosmetic compositions and a massage oil comprising a rosewood essential oil obtained by supercritical CO2 without stating the parameters applied for the extraction.

[0009] The rosewood extract according to the present invention has been found to be useful in preventing or limiting the signs of skin aging and in particular in firming the skin, limiting the appearance of facial ptosis, redefining the contour of the face or limiting the appearance of wrinkles and fine lines. Summary of the invention

[0010] The first object of the invention is to obtain an extract obtained from leaves or shavings of trunks and branches of rosewood (Aniba rosaeodora) previously exhausted, according to the process comprising the following steps:

[0011] a) between 5 and 25% water is added to the exhausted and dried rosewood;

[0012] b) an extraction is carried out using a fluid in a supercritical state such as carbon dioxide (CO2) in the presence of a polar co-solvent, chosen from primary or secondary alcohols, or any mixture thereof;

[0013] c) a support solvent chosen from polyol type solvents, saturated or unsaturated, linear or branched fatty alcohols, comprising from 8 to 30 carbons, glyceride type solvents or long carbon chain ester type solvents, or any mixture thereof;

[0014] d) the extract solubilized in step c) is evaporated to remove all of the co-solvent, ISP‐BV23‐200PCT Description

[0015] e) the extract is filtered to remove the precipitated compounds which do not dissolve in the carrier solvent and to collect the filtrate,

[0016] f) the filtered extract obtained in e) is diluted in the same carrier solvent at a concentration of between 0.005 and 50% of crude extract by weight of the total weight of the final extract.

[0017] The second subject of the invention is a purified extract of exhausted rosewood capable of being obtained by the process according to the invention, characterized in that it is diluted to a concentration of between 0.1 and 1% in the carrier solvent and comprises from 0.3 to 0.5%, advantageously from 0.35 to 0.5% and preferably between 0.4 and 0.5% of semi- to non-volatile compounds.

[0018] The third subject of the invention is a composition comprising, as active agent, an effective amount of a purified extract of rosewood, obtained according to the process of the invention, and a physiologically acceptable medium.

[0019] The fourth subject of the invention is the cosmetic use of a composition comprising a purified extract of rosewood to prevent or limit the signs of skin aging, to firm the skin, limit the appearance of facial ptosis, redefine the contour of the face or even limit the appearance of wrinkles and fine lines.

[0020] The invention also relates to a composition comprising a purified extract of rosewood for use in obtaining an effect similar to that of retinoids, without exhibiting undesirable inflammatory effects. Brief description of the figures

[0021] The following figures illustrate the advantages which arise from the invention and the non-limiting embodiments which are presented in the description:

[0022] [Fig.1] Analytical comparison by HS-GC / MS of the volatile compound composition of spent rosewood dregs and rosewood essential oil as described in Example 3.

[0023] [Fig.2] Analytical comparison by HPLC / UV (300 nm) of the composition in semi-to non-volatile compounds of a rosewood essential oil as described in Example 3 and of the purified rosewood extract according to Example 1.

[0024] [Fig.3] Analytical comparison by HS-GC / MS of the purified rosewood extract according to Example 1 and a rosewood essential oil as described in Example 3. ISP‐BV23‐200PCT Description

[0025] [Fig.4] Analytical comparison by HPLC / DEDL of the crude rosewood extract and the purified rosewood extract according to Example 1 describing the difference in anibine content.

[0026] [Fig.5] Evaluation of 1% purified rosewood extract on the expression of type I, type III collagen and hyaluronic acid in human skin biopsies.

[0027] [Fig.6] Evaluation of 1% purified rosewood extract on CRABP2 expression in human skin biopsies compared to Retinoic acid.

[0028] [Fig.7] Evaluation of 1% purified rosewood extract on the expression of the alpha 3 chain of type V collagen on human skin biopsies in comparison with Retinoic acid.

[0029] [Fig.8] Evaluation of the effect of a 1% purified rosewood extract on the volume of the facial oval (a) and the angle V of the facial oval (b).

[0030] [Fig.9] Evaluation of the effect of a purified 1% rosewood extract on the distance of facial ptosis (a) and on the volume of ptosis (b).

[0031] [Fig.10] Evaluation of the effect of a 1% purified rosewood extract on lip corner volume (a) and lip corner profile roughness Ra (b) and Rz (c).

[0032] [Fig.11] Evaluation of the effect of a purified 1% rosewood extract on the profile roughness Ra (a) and Rz (b) of crow's feet wrinkles.

[0033] [Fig.12] Evaluation of 1% purified rosewood extract on the expression of interleukin 1 receptor type 1 (IL1R1) on human skin biopsies in comparison with Retinoic acid. Detailed description of the invention Definitions

[0034] All terms used in this specification have the most widely known meanings unless otherwise stated. For the purposes of the invention the following terms are defined as follows:

[0035] In this description, the expression "rosewood" or "rosewood (Aniba rosaeodora)" or "exhausted rosewood" or even "co-products or by-products of the distillation of rosewood essential oil" means, without distinction, the dregs of chips from the trunk, branches or leaves of rosewood (Aniba rosaeodora) recovered after ISP‐BV23‐200PCT Description essential oil extraction. The essential oil content in the fresh plant varies between 0.4 and 3.5%, including an average linalool content of between 0.32 and 2.8%. Essential oil is traditionally extracted by hydrodistillation or steam distillation. The term "exhausted" therefore refers to plant material that has undergone a first extraction treatment, such as steam distillation or hydrodistillation, or any solid / liquid extraction processes, aimed at extracting between 80 and 95% of the volatile compounds present in this raw material.

[0036] Or, the term "exhausted rosewood" refers to rosewood containing a maximum of 20% of the volatile compounds present in the initial raw material.

[0037] The term "rosewood trunk and branches" means the parts of the rosewood corresponding to the trunk and branches with bark.

[0038] A "polar co-solvent" means a solvent with a polarity greater than that of CO2 in the supercritical state, such as primary or secondary alcohols, or any mixture thereof.

[0039] “Polyol type” solvent means organic compounds having at least 2 hydroxyl groups, such as diols or triols, or a mixture of these compounds.

[0040] “Fatty alcohol type” solvent means organic compounds, saturated or unsaturated, linear or branched, possessing a hydroxyl group, or a mixture of these compounds.

[0041] A "glyceride-type" solvent is defined as fatty acid esters and glycerol, or a mixture of these compounds.

[0042] “Long carbon chain ester type” solvents are understood to mean organic compounds having an ester function substituted by long carbon chains.

[0043] The term “support solvent” means a solvent chosen from polyol type solvents, saturated or unsaturated fatty alcohols, linear or branched, comprising from 8 to 30 carbons, glyceride type solvents or long carbon chain ester type solvents, or any mixture thereof.

[0044] "Volatile compounds" are organic compounds that can easily pass into the gaseous phase at atmospheric pressure and room temperature.

[0045] “Semi- to non-volatile compounds” are compounds that do not readily pass into the gas phase at atmospheric pressure and room temperature due to: ISP‐BV23‐200PCT Description of a higher boiling point than in the case of volatile molecules. In the context of the invention, these are moderately polar molecules, mainly belonging to the following chemical families: vitamins, aryl phenyl ketones, sesquiterpenes, sesquiterpene derivatives and phytosterols.

[0046] The term "purified rosewood extract" means the extract in liquid form purified by partial precipitation of anibine, after dilution of the crude extract in a solvent of the polyol type, saturated or unsaturated fatty alcohol, linear or branched, comprising from 8 to 30 carbons, solvents of the glyceride or ester type with a long carbon chain, or any mixture thereof, and filtered to remove the precipitate.

[0047] The term “raw extract” of rosewood means the extract in pasty form obtained by steps a), b) and c') of the process by complete evaporation of the co-solvent without adding a carrier solvent.

[0048] When a range of values ​​is described, the bounds of that range should be understood to explicitly include the upper and lower bounds of that range, as well as all intermediate values ​​within the range. For example, a range of values ​​between 1% and 10% should be understood to include 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, as well as all decimal values ​​between 1% and 10%.

[0049] Numerical percentage values ​​are percentages by weight, i.e. the weight of a compound relative to the total weight of the intended mixture, unless otherwise specified.

[0050] The compositions described herein may "comprise", "consist of" or "consist essentially of", the essential compounds or the optional ingredients.

[0051] “Consist essentially of” means that the composition or component may include additional ingredients, but only if the additional ingredients do not alter the basic characteristics or novel characteristics of the composition or use described in this application.

[0052] “Physiologically acceptable medium” means a solvent suitable for contact with the outer layers of the skin, scalp or appendages, without toxicity, irritation, undue allergic response or the like or reaction ISP‐BV23‐200PCT Description of intolerance, and proportionate to a reasonable benefit / risk ratio, at the concentrations used.

[0053] By “effective quantity” we mean the minimum quantity of extract according to the invention which is necessary to obtain at least one of the desired biological activities.

[0054] Facial ptosis refers to the sagging associated with the sliding and atrophy of fatty tissue and the loss of facial bone support with age. Extraction procedures

[0055] Rosewood has been overexploited for its essential oil, placing it in the category of species protected by the IUCN (International Union for Conservation of Nature) whose trade is regulated by the Convention on International Trade in Endangered Species (CITES).

[0056] The inventors therefore chose to use co-products from rosewood distillation, in order to reduce the impact on the environment and on biodiversity and allow better management of plant resources.

[0057] The leaves and shavings of the trunk or branches of rosewood (Aniba rosaeodora) recovered after extraction of the essential oil by distillation are dried so that the residual moisture content in the plant material is between 1 and 10%.

[0058] The objective of the process of the invention is to concentrate and promote the chemical diversity of the extract, the content of semi- to non-volatile compounds while maintaining the highest possible extraction yield.

[0059] The extraction is carried out using supercritical fluid such as supercritical CO2, which is an alternative process to distillation that meets the principles of green chemistry. The addition of a polar co-solvent to supercritical CO2 increases the polarity of the CO2 and therefore extracts chemical families of metabolites over a wider range of polarity, such as carbohydrates, glycosylated or non-glycosylated flavonoids, phenolic acids, triterpenes, lipids, phytosterols.

[0060] A screening experimental design made it possible to highlight the most influential parameters on the following criteria: extraction yield, chemical diversity of the extract and the content of semi- to non-volatile compounds. The parameters studied were grinding, the humidification rate of the plant material, the addition or not of ISP‐BV23‐200PCT Description inert compound such as cellulose powder, co-solvent flow rate, and extraction temperature and pressure.

[0061] The screening experiment design surprisingly highlighted that the co-solvent flow rate and the humidification rate of the plant material were the two parameters that exerted an influence on the extraction, unlike the other parameters which did not show a significant impact.

[0062] These two influential parameters were then more finely analyzed by an optimization experiment plan applying the response surface method.

[0063] Finally, the support solvent added before the evaporation of the co-solvent was selected following numerous tests carried out to determine the best solvent allowing both the concentration of the compounds of interest and also the maximum elimination of anibine by precipitation.

[0064] Anibine is a substance classified as an irritant, and belongs to a chemical class known to possess marked biological activity often associated with some toxicity.

[0065] The invention thus has as its first object a process for obtaining a rosewood extract obtained from leaves or shavings of trunks and branches of exhausted rosewood (Aniba rosaeodora) comprising the following steps:

[0066] a) between 5 and 25% water is added to the exhausted and dried rosewood;

[0067] b) an extraction is carried out using a fluid in a supercritical state such as carbon dioxide (CO2) in the presence of a polar co-solvent, chosen from primary or secondary alcohols, or any mixture thereof;

[0068] c) a support solvent chosen from polyol type solvents, saturated or unsaturated, linear or branched fatty alcohols, comprising from 8 to 30 carbons, glyceride type solvents or long carbon chain ester type solvents, or any mixture thereof;

[0069] d) the extract solubilized in step c) is evaporated to remove all of the co-solvent,

[0070] e) the extract is filtered to remove the precipitated compounds which do not dissolve in the carrier solvent and to collect the filtrate,

[0071] f) the filtered extract obtained in e) is diluted in the same carrier solvent at a concentration of between 0.1 and 1% of crude extract by weight of the total weight of the final extract. ISP‐BV23‐200PCT Description

[0072] In step a), dried grains from the distillation of trunk and branch chips are advantageously used, to which between 5 and 25% of water is added to the exhausted and dried rosewood to obtain an optimum humidification rate.

[0073] Advantageously, the exhausted chips can come from the trunk and branches of rosewood grown in Latin America.

[0074] Preferably, prior to step a), the raw material is ground in the presence or absence of liquid nitrogen to obtain a coarse-grained powder.

[0075] In step b) preferably the co-solvent is ethanol at a concentration of between 80 and 100% (volume / volume in water), preferably between 90 and 100% and even more preferably at a concentration of 96% in water (volume / volume).

[0076] Preferably, the mass ratio of carbon dioxide (CO2) in the supercritical state, relative to the quantity of moistened raw material used, is between 10 and 50, advantageously between 20 and 40 and preferably between 25 and 35.

[0077] Preferably, the mass ratio of the co-solvent to the supercritical solvent (carbon dioxide) is between 0.050 and 0.080, advantageously between 0.055 and 0.075 and preferably between 0.060 and 0.070.

[0078] Preferably the flow rate of co-solvent; when this is ethanol; is between 10 and 20 ml / min. Even more preferably, the flow rate of the co-solvent is set at 20 ml / min.

[0079] Preferably, the extraction temperature is between 35 and 85°C, advantageously between 45 and 75°C and preferably between 55 and 65°C.

[0080] Preferably, the pressure within the extractor is between 90 and 1000 bar, preferably between 150 and 700 bar and even more preferably between 250 and 350 bar.

[0081] To carry out step b) advantageously, the mixture obtained in step a) is placed in a stainless steel cartridge, this cartridge is introduced into a supercritical fluid extractor.

[0082] The presence of a polar co-solvent in the extraction conditions described above allows the production of an extract with notable phytochemical diversity in ISP‐BV23‐200PCT Description that it contains both a residual portion of volatile compounds not extracted during distillation and semi- to non-volatile compounds of variable polarity belonging to the following chemical families: vitamin and derivative, aryl phenyl ketones, sesquiterpene derivatives, sesquiterpenes, phytosterols and free fatty acids.

[0083] In step c), preferably, the quantity of solvent called "support" necessary to obtain a crude extract content of between 5 and 80% by weight of the total weight of the final extract, advantageously between 10 and 50% and preferably between 15 and 25%, is added to the extract obtained in step b).

[0084] Preferably the carrier solvent is selected from octyldodecanol, 2-hexyl decanol, oleyl alcohol, glycerol tricaprylate / caprate or an octyldodecyl stearoyl stearate ester, or any mixture thereof.

[0085] Even more preferably, the carrier solvent is glycerol tricaprylate / caprate.

[0086] In the presence of the carrier solvent, precipitation is observed. Analysis shows that the precipitate is mainly composed of anibine, as illustrated by the chromatographic profiles in Figure 4.

[0087] In step d) the optimal conditions for evaporation of the co-solvent are an evaporation temperature of at most 60°C and a pressure of less than 60 mbar.

[0088] In step e) the liquid extract obtained is filtered to remove the precipitated anibine. The filtrate is collected.

[0089] In step f) the filtrate is preferably diluted to a concentration of 0.5% by weight of crude extract / weight of final extract.

[0090] Advantageously, the process can be continued by at least one optional purification step g), according to any technique known to those skilled in the art and in particular by chromatography or by molecular distillation.

[0091] Alternatively, at the end of step b), it is possible to implement another extraction process by carrying out a step c') of total evaporation of the co-solvent, without adding a support solvent. A crude extract is then obtained in pasty form. This extract can be used, among other things, for analytical chemistry studies which could be disturbed by the presence of a support solvent.

[0092] At the end of step c') the extraction yield is between 0.5 and 5%. ISP‐BV23‐200PCT Description Extract

[0093] The second subject of the invention is a purified extract of exhausted rosewood capable of being obtained by the process according to the invention, characterized in that it is diluted to a concentration of between 0.1 and 1% in the carrier solvent and comprises from 0.3 to 0.5%, advantageously from 0.35 to 0.5% and preferably between 0.4 and 0.5% of semi- to non-volatile compounds.

[0094] The purified rosewood extract is advantageously solubilized in glycerol tricaprylate / caprate.

[0095] In another embodiment the purified rosewood extract comprises 0.5% crude extract, by weight of the total weight of the final extract.

[0096] In this particular embodiment, the semi- to non-volatile compounds comprise from 0.01 to 0.4% of sesquiterpene derivatives in cotoine equivalent, from 0.001 to 0.1% of aryl phenyl ketones including 0.001 to 0.05% of cotoine, from 0.001 to 0.01% of beta-sitosterol and from 0.001 to 0.1% of anibine.

[0097] As an example, we can cite purified extract of rosewood diluted to 0.5% in glycerol tricaprylate / caprate by weight of crude extract relative to the weight of the final extract and comprising 0.08% of sesquiterpene derivatives in cotoine equivalent, 0.006% of aryl phenyl ketones including 0.005% of cotoine and 0.009% of anibine.

[0098] The extract of the invention is different from the rosewood extracts described in the prior art, and in particular from the essential oils, in that it has a much less intense olfactory note, a significantly reduced content of volatile and allergenic compounds, and semi- to non-volatile compounds of variable polarity belonging to several chemical families.

[0099] In the purified rosewood extract of the invention, volatile compounds are present at concentrations below the detection limit and therefore cannot be quantified. The linalool concentration is less than 0.01%.

[0100] The phytochemical study to characterize the compounds was carried out on the crude extract of exhausted rosewood, obtained by a process comprising steps a), b) and c').

[0101] Raw Rosewood Extract includes: ISP‐BV23‐200PCT Description

[0102] Between 5 and 20% of volatile compounds, mainly sesquiterpenes and sesquiterpene alcohols, preferably between 8 and 18%, even more preferably 10 and 15% of volatile compounds and a maximum of 1% of linalool.

[0103] Between 80 and 95% of semi- to non-volatile compounds, preferably between 82 and 92%, and even more preferably between 85 and 90%. This fraction is mainly composed of sesquiterpene derivatives (approximately 30% of the extract), aryl phenyl ketones (approximately 10% of the extract including approximately 5% cotoine), a nicotinic acid derivative (approximately 20% of the extract), sesquiterpenes such as eudesmane and guaiane, nicotinic acid, organic acids, as well as more apolar molecules belonging to the chemical families of the phytosterol and fatty acid type (approximately 25% of the extract for all these other compounds.

[0104] In a very advantageous embodiment, the crude rosewood extract comprises 13% volatile compounds and 87% semi- to non-volatile compounds including 32% sesquiterpene derivatives, 12% aryl phenyl ketones including 6% cotoine, 20% anibine and 23% sesquiterpene compounds, free fatty acids and phytosterols.

[0105] The non-exhaustive list of compounds present in these two fractions analyzed by HPLC coupled to a diode array detector or an evaporative light scattering detector by external calibration is given as an example in the following table 1:

[0106] [Table 1] Fraction Chemical family Identification ISP‐BV23‐200PCT Description Alpha‐dictyopterol Beta‐dictyopterol

[0007] Some markers could be quantified in the raw rosewood extract. This contains in particular:

[0108] Between 10 and 50% of sesquiterpene derivatives, advantageously between 20 and 40% and preferably between 25 and 35%;

[0109] Between 1 and 25% of aryl phenyl ketones, advantageously between 5 and 20% and preferably between 10 and 15% of which between 5 and 10% of cotoine;

[0110] Between 0.01 and 2% beta-sitosterol, advantageously between 0.1 and 1.5% and preferably between 0.5 and 1%;

[0111] Between 1 and 40% anibine, advantageously between 5 and 30% and preferably between 10 and 20%. Compositions

[0112] A third subject of the invention is a composition comprising as active agent, an effective amount of a purified extract of rosewood, obtained according to the process described above, and a physiologically acceptable medium.

[0113] Advantageously, the purified rosewood extract is added to a physiologically acceptable medium at a concentration of between 0.01% and 10% of purified rosewood extract by weight relative to the total weight of the composition, preferably at a concentration of between 0.1 and 5% by weight relative to the total weight of the composition, and even more preferably at a concentration of between 0.5 and 2% by weight relative to the total weight of the composition.

[0114] The composition of the present application is formulated to be applied by any suitable route, in particular orally, or externally, and the formulation of the compositions will be adapted by those skilled in the art.

[0115] Preferably, the composition of the present application is in a form suitable for topical application. This composition must therefore contain a physiologically acceptable medium, that is to say compatible with the skin and appendages, without risk of discomfort during its application. ISP‐BV23‐200PCT Description

[0116] The composition may in particular be presented in the form of an aqueous, hydroalcoholic or oily solution or gel, an oil-in-water emulsion, water-in-oil or multiple emulsions; they may also be presented in the form of suspensions, or even powders, suitable for application to the skin, mucous membranes, lips and / or hair.

[0117] The composition can be more or less viscous and also have the appearance of a cream, lotion, fluid, milk, serum, ointment, gel, paste, balm or mousse. It can also be in solid form, such as a stick or be applied to the skin in the form of an aerosol.

[0118] As a physiologically acceptable medium commonly used in the intended field of application, mention may be made, for example, of adjuvants necessary for the formulation, such as solvents, thickeners, gelling agents, diluents, emulsifiers, antioxidants, colorants, sunscreens, self-tanning agents, pigments, fillers, preservatives, perfumes, odor absorbers, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, esters, vegetable oils or butters, etc.

[0119] In all cases, the person skilled in the art will ensure that these adjuvants and their proportions are chosen in such a way as not to harm the desired advantageous properties of the composition according to the invention.

[0120] According to another advantageous embodiment, the purified rosewood extract can be encapsulated or included in a cosmetic vector such as liposomes or any other nanocapsule or microcapsule used in the field of cosmetics or adsorbed on powdery organic polymers, mineral supports such as talcs and bentonites.

[0121] Advantageously, the composition may comprise, in addition to the active agent, that is to say a purified extract of rosewood, at least one other active agent having cosmetic effects similar and / or complementary to those of the invention.

[0122] Such additional active agents may also be selected according to their chemical composition, from the group comprising: amino sugars, glucosamine, D-glucosamine, N-acetyl-glucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetyl mannosamine, galactosamine, N-acetyl galactosamine, vitamin B3 and its derivatives, niacinamide, sodium dehydroacetate, dehydroacetic acid and its salts, phytosterols, salicylic acid compounds, hexamidines, dihydroxyproline compounds of ISP‐BV23‐200PCT Description dialkanoyl, soy extracts and derivatives, equol, isoflavones, flavonoids, phytantriol, farnesol, geraniol, bisabolol, peptides and their derivatives, di-, tri-, tetra-, penta-, and hexapeptides and their derivatives, lys-thr-thr-lys-ser, palmitoyl-lys-thr-thr-lys-ser, carnosine, N-acyl amino acid compounds, retinoids, retinyl propionate, retinol, retinyl palmitate, retinyl acetate, retinal, retinoic acid, water-soluble vitamins, ascorbates, 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 Bl, vitamin B2, vitamin B6, vitamin B12, vitamin E, vitamin F, vitamin K and its derivatives, pantothenic acid and its derivatives, pantothenyl ethyl ether, panthenol and its derivatives, ethyl panthenol, dexpanthenol, biotin,amino acids and their salts and derivatives, water-soluble amino acids, asparagine, alanine, indole, glutamic acid, water-insoluble vitamins, beta-ionol, cedrol, and their derivatives, water-insoluble amino acids, tyrosine, tryptamine, particulate materials, butylated hydroxytoluene, butylated hydroxyanisole, allantoin, tocopheryl nicotinate, tocopherol, tocopherol esters, palmitoyl-gly-his-lys, phytosterol, hydroxy acids, glycolic acid, lactic acid, lactobionic acid, keto acids, pyruvic acid, phytic acid, lysophosphatidic acid, stilbenes, cinnamates, resveratrol, kinetin, zeatin, dimethylaminoethanol, natural peptides, soy peptides, acid sugar salts, manganese gluconate, zinc gluconate, piroctone olamine, 3,4,4'- trichlorocarbanilide, triclocarban, zinc pyrithione, hydroquinone, kojic acid, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl glucoside, pyridoxine, aloe vera, terpene alcohols, allantoin,bisabolol, dipotassium glycyrrhizate, glyceryl acid, sorbitol, pentaerythritol, pyrrolidone and its salts, dihydroxyacetone, erythrulose, glyceraldehyde, tartaraldehyde, clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, witch hazel distillate, eicosene and vinyl pyrrolidone copolymer, iodopropyl butylcarbamate, a polysaccharide, an essential fatty acid, a salicylate, a glycyrrhetinic acid, carotenoids, ceramides and pseudoceramides, a complex lipid, oils generally of natural origin such as shea butter, apricot oil, evening primrose oil, prune oil, palm oil, oil monoi, kahai oil, hydroquinone, HEPES, procysteine, O-octanoyl-6-D-maltose, methyl glycine diacetic acid disodium salt, steroids such as diosgenin and DHEA derivatives, DHEA dehydroepiandrosterone and / or a chemical or biological precursor or derivative,N-ethylcarbonyl-4-para-aminophenol, alpha hydroxy acids, beta, ISP‐BV23‐200PCT Description hydroxy acids, moisturizers, epidermal hydrolytic enzymes, plant extracts, phytohormones, yeast extracts, a metalloproteinase inhibitor, enzymes, enzyme inhibitors, enzyme inducers, coenzymes, chelating agents, plant extracts and plant derivatives, essential oils, marine extracts, agents from a biofermentation and / or biotechnology process, mineral salts, cell extracts.

[0123] Examples include:

[0124] the peptides commercially known as MATRIXYL®, ARGIRELINE®, CHRONOGEN™, LAMINIXYL IS™, PEPTIDE Q10™, COLLAXYL™ (patent FR2827170, ASHLAND®), PEPTIDE VINCI 01™ (patent FR2837098, ASHLAND®), PEPTIDE VINCI 02™ (patent FR2841781, ASHLAND®), ATPeptide™ (patent FR2846883, ASHLAND®) or the synthetic peptide with the sequence Arg-Gly-Ser-NH2, marketed under the name ATPeptide™ by ASHLAND®;

[0125] Artemia salina extract, marketed under the name GP4G™ (FR2817748, ASHLAND®);

[0126] plant peptide extracts such as flax extracts (Lipigenin™, patent FR2956818, ASHLAND®), soybean, spelt, vine, rapeseed, flax, rice, corn, pea, cocoa extracts;

[0127] yeast extracts, for example Dynagen™, (patent FR2951946, ASHLAND®) or Actopontine™ (patent FR2944526, ASHLAND®). Uses

[0128] The fourth subject of the invention is the cosmetic use of a composition described above comprising a purified extract of rosewood to prevent or limit the signs of skin aging and in particular to firm the skin, limit the appearance of facial ptosis, redraw the contour of the face, reshape and restructure the face and limit the appearance of wrinkles and fine lines.

[0129] The term "cosmetic use" means that the use is intended for individuals with healthy skin, scalp or appendages.

[0130] By "signs of skin aging" we mean any change in the external appearance of the skin due to aging, such as, for example, sagging of the face with a loss of the oval contour of the face, the appearance of ptosis or jowls, a loss of volume in the cheeks and cheekbones, the appearance of wrinkles and fine lines at the corners ISP‐BV23‐200PCT Description of lips and crow's feet, wilting, loss of elasticity, firmness and / or tone of the skin, but also any internal changes to the skin that do not systematically result in a modified external appearance such as, for example, thinning of the skin, or any internal damage to the skin resulting from external aggressions producing free radicals, such as pollution and solar radiation including UV rays.

[0131] Facial shape is genetically determined and it is known that a large number of genetic loci influence facial shape and contours.

[0132] The extracellular matrix is ​​one of the primary determinants of facial morphology, so rosewood extract was tested on the expression of collagens I and III, and hyaluronic acid and showed a positive effect on the expression of these proteins.

[0133] The inventors have now identified, through bioinformatics analysis of genome-wide association study data, certain genetic determinants involved in facial shape and contours. These genetic determinants include, among others, collagen XVII, fibulin-7, and harmonin.

[0134] Fibulin-7, in addition to its involvement in facial shape and contours, is thought to interact with the extracellular matrix, and studies have suggested that it plays a crucial role in maintaining epidermal stem cells during skin aging.

[0135] Collagen XVII is a transmembrane protein found in hemidesmosomes (HD). Collagen XVII is involved in stem cell interactions with surrounding cells and the matrix and may also support the maintenance of stem cells and their niches. Some mutations responsible for collagen XVII deficiency lead to premature hair graying phenotypes.

[0136] Harmonin is a scaffolding protein, a family of proteins known to play a central role in macromolecular assemblies.

[0137] Rosewood extract was tested on the expression of fibulin-7 and collagen XVII, and harmonin and showed a positive effect on the expression of these proteins, in relation to the anti-wrinkle and restoration effect of the shape and contours of the face observed in a clinical test.

[0138] Genetic variations in the alpha 3 chain of type V collagen have been described in a panel of Asian volunteers as being associated with ISP‐BV23‐200PCT Description genetic predispositions related to skin sagging and wrinkles in the corner of the eye (Okuno R. et al., Exp Dermatol.2022;31(9):1411-1420.). In addition, a deficiency in the alpha 3 chain of type V collagen induced by the action of matrix metalloproteinase 9 (MMP-9) would be significantly associated with eczema (Martin MJ et al. Genes (Basel). 2020;11(4):442).

[0139] The rosewood extract of the invention was tested on the expression of the alpha 3 chain of type V collagen, and showed a positive effect on the expression of this protein, which suggests an anti-aging effect particularly interesting for Asian and Caucasian skin and in particular for Asian skin.

[0140] The invention also relates to the cosmetic use of the composition of the invention to increase the expression of the following proteins: collagen XVII, fibulin-7, harmonin, alpha 3 chain of type V collagen and CRABP2.

[0141] The set of genes that code for these proteins can be considered a "Facial Architecture Genomic Complex" or "Facial Architectural Genomic Complex".

[0142] Vitamin A and its derivatives, especially retinol, are among the most effective substances for delaying the skin aging process. Similarly, retinoids, which are naturally occurring or synthetic derivatives of vitamin A, are widely used in dermatology in anti-aging treatments (Mukherjee S, et al. Retinoids in the treatment of skin aging: an overview of clinical efficacy and safety. Clin Interv Aging. 2006;1(4):327-348) and their effectiveness on photoaging and restructuring of the dermis and epidermis are well described. However, it is also known that these molecules have drying and irritating side effects.

[0143] Rosewood extract was tested on the expression of an inflammation marker: the interleukin 1 receptor (IL1R1), and showed no significant modulation on the expression of this protein. In comparison, retinoic acid caused an increase in the expression of this protein.

[0144] CRABP1 and CRABP2, the retinoic acid-specific binding proteins, bind to retinoic acid and promote its transport within the cell. Deficiency in CRABP2 expression is known to accelerate skin aging (Bielli A. et al., Aging (Albany NY). 2019;11(6):1619-1632. ISP‐BV23‐200PCT Description

[0145] Rosewood extract was tested on CRABP2 expression and showed a positive effect on the expression of this protein and did not increase IL1R1 expression. Purified rosewood extract shows comparable effects to retinoic acid on CRABP2 expression, and has the advantage of not causing inflammatory side effects, which most sensitive skin cannot tolerate.

[0146] The composition of the invention has good tolerance, which allows its use on sensitive skin or skin with characteristics of atopic dermatitis.

[0147] The subject of the invention is thus a composition described in the present invention for its use in obtaining an effect similar to that of retinoids, without exhibiting undesirable inflammatory effects (“retinol-like” effect).

[0148] The invention also relates to the cosmetic use of the composition of the invention to obtain an effect similar to that of retinoids on the signs of skin aging, without exhibiting undesirable inflammatory effects.

[0149] The invention also relates to a composition comprising a purified extract of rosewood for use in combating external attacks producing pro-inflammatory free radicals, such as stress, pollution or external attacks. Examples

[0150] The present invention will now be illustrated by means of the following non-limiting examples: Example 1: Preparation of a crude extract and a purified extract of exhausted rosewood

[0151] For the realization of the examples, trees of the species Aniba rosaeodora were cultivated in Peru.

[0152] The parts of the plant used are the spent grains of shavings of trunk and branches of the Aniba rosaeodora tree, exhausted of the essential oil (volatile compounds) by a first distillation process by steam distillation. During the development of the process of the invention, the extraction parameters were optimized so as to extract the maximum of semi-to non-volatile compounds with maximum yield. The first stage of the development consisted of highlighting the factors which had the most influence on the extraction. Thus, several parameters were studied namely the interest of cryogenic grinding, the humidification rate of the plant material, the ISP‐BV23‐200PCT Description percentage of cellulose added, co-solvent flow rate, as well as temperature and pressure in the extractor. Only factors that showed a real impact on extraction were optimized within the framework of an optimization experiment design applied according to the response surface method.

[0153] Table 2 shows some simulated results obtained by the experimental design carried out to optimize the extraction parameters:

[0154] [Table 2] Peak area Peak area Flow rate co- Humidification Efficiency DEDL DEDL [ d exp rence are a cotone and a rv ea cotone. aque are act corrg e by the concentration of the solution injected in HPLC in order to free itself from variations linked to sample preparation. ISP‐BV23‐200PCT Description

[0156] The extraction conditions were selected to achieve a satisfactory extraction yield while ensuring the extract had maximum content and chemical diversity in semi- to non-volatile compounds. The preferred extraction parameters are as follows:

[0157] The exhausted rosewood chips are cryogenically ground using a 4 mm sieve. The resulting plant powder is moistened with 15% water (% by mass). The humified plant material is introduced into a stainless steel cartridge. This cartridge is placed in a supercritical fluid extractor such as the Separex SFE 5 extractor. The extraction solvent used is supercritical carbon dioxide at a flow rate of 15 kg / h with the addition of a polar co-solvent, corresponding to 96% ethanol in water (volume / volume) at a flow rate of 20 ml / min. The mass ratio of carbon dioxide to moistened plant material is 30 and that of the co-solvent to carbon dioxide is 0.065. The pressure and temperature within the extractor are 300 bar and 60°C respectively. The temperature within the separator is 35°C. A measurement of the dry extract is carried out on the ethanolic extract obtained for the preparation of the purified extract.

[0158] In the case of the crude extract, the solution of ethanolic extract of exhausted rosewood is evaporated under vacuum until the ethanol has completely evaporated (pressure less than 90 mbar and water bath temperature at 60°C maximum). An extract is then obtained in the form of a relatively solid paste.

[0159] To prepare the purified extract, before evaporation under vacuum, the supporting solvent, namely glycerol tricaprylate / caprate, is added to the ethanolic extract solution so as to obtain a diluted solution of 20% crude extract by weight of the total weight of the diluted extract. The mixture is evaporated under vacuum until the ethanol has completely evaporated (pressure less than 90 mbar and water bath temperature of 60°C maximum). In the presence of glycerol tricaprylate / caprate, anibine partially precipitates.

[0160] The extract is filtered and the precipitate removed.

[0161] The glycerol tricaprylate / caprate content in the obtained filtrate is quantified by HPLC. From this result, the extract is diluted in the same solvent (glycerol tricaprylate / caprate) to obtain a clear, fluid extract containing 0.5% of crude rosewood extract.

[0162] The purified extract obtained is in the form of a liquid extract. ISP‐BV23‐200PCT Description

[0163] Several markers of the extract were quantified. Thus, the purified rosewood extract is 0.5% in glycerol tricaprylate / caprate (by weight of crude extract relative to the weight of the final extract) and comprises 0.08% sesquiterpene derivatives in cotoine equivalent, 0.006% aryl phenyl ketones including 0.005% cotoine, 0.004% beta-sitosterol, as well as 0.009% anibine. The linalool content is below the detection limit, i.e. a content lower than 0.01%. Example 2: Characterization of the crude rosewood extract obtained according to Example 1

[0164] The phytochemical study was carried out on the crude extract of exhausted rosewood. This extract is obtained under the conditions cited in Example 1 in the case where the co-solvent (ethanol) is completely evaporated under vacuum (90 mbar, water bath temperature at 60°C maximum) without the addition of a carrier solvent. The crude extract is then obtained in a relatively solid pasty form.

[0165] Volatile compounds are analyzed by gas chromatography (GC) coupled with mass spectrometry (MS) and / or flame ionization detection (FID). Confirmation of identifications is made possible by comparison of linear retention indices and mass spectra contained in libraries. Quantification by GC / FID is done by internal calibration using predicted and / or calculated response factors.

[0166] Semi- to non-volatile compounds are monitored by high-performance liquid chromatography (HPLC) coupled with a UV diode array detector (DAD) and an evaporative light scattering detector (ELSD). Structural identifications are confirmed by nuclear magnetic resonance (NMR) experiments as well as high-resolution mass spectrometry (HRMS) analyses. Identification is also confirmed by injection of the standard if it is commercially available.

[0167] Sesquiterpene derivatives, aryl phenyl ketone compounds and beta-sitosterol were quantified by HPLC. Cotoine is used as an external standard for the determination of sesquiterpene derivatives and aryl phenyl ketones. The content is then expressed as cotoine equivalent. The analytical method developed for this determination has been subject to method validation according to the guidelines indicated by the ICH (International Conference on Harmonization)

[0168] The non-exhaustive list of compounds present in the volatile fraction and the semi-to-non-volatile fraction is detailed in Table 3 below: ISP‐BV23‐200PCT Description

[0169] [Table 3] Chemical Family Identification ISP‐BV23‐200PCT Description Sitosterol [0 this ex trat. content years:

[0171] 13% volatile fraction; of which a maximum of 1% linalool

[0172] 87% semi-to-non-volatile fraction including 32% sesquiterpene derivatives, 12% aryl phenyl ketones including 5% cotoine, 20% anibine and 23% various compounds such as sesquiterpenes, free fatty acids, phytosterols and other lipid compounds.

[0173] Example 3: Production of a rosewood extract of the essential oil type

[0174] In order to carry out a comparative analysis, an extraction of rosewood essential oil (not exhausted) was carried out in a conventional manner, by steam distillation. The trunk and branches of Aniba rosaeodora are dried and then roughly crushed to give chips which are placed in separate stills to then be crossed by a stream of water vapor; this steam releases the volatile molecules or essential oil which is carried away by the water vapor and condenses in the condenser. Since the essential oil is less dense than water and is not or only slightly water-soluble, it is collected at the outlet in a decanter called an essencier. The water which still contains essential oil in trace amounts is called hydrosol.

[0175] The essential oil is a colorless to pale yellow liquid (density 0.8789, refractive index 1.4644, optical rotation +1.15). Example 4: Demonstration of the exhaustion of the raw material by steam distillation

[0176] The rosewood raw material used in the context of the invention is said to be “exhausted”. This means that it has undergone a first extraction treatment, in this case steam distillation, which has made it possible to extract the volatile fraction of the rosewood, i.e. the essential oil.

[0177] Spent rosewood grains were analyzed by gas chromatography-mass spectrometry (GC / MS) and flame ionization detector (FID). Injection was performed in headspace. ISP‐BV23‐200PCT Description This sample preparation favors the analysis of the most volatile compounds. The separation was obtained on a 30 x 0.25 m, 0.25 µm apolar column, with elution according to a temperature gradient (isotherm 50°C, 5 min; then 3°C / min up to 120°C; then 5°C / min up to 250°C; isotherm 250°C, 5 min). The carrier gas used is helium.

[0178] Figure 1 shows the GC / MS chromatographic profile of spent rosewood grains compared to the GC / MS chromatographic profile of rosewood essential oil. It is noted that the essential oil is much richer in volatile compounds. These compounds are eluted before 40 min under the conditions of the method described above. The abundance of peaks is also much more intense than in the case of rosewood grains.

[0179] The chromatographic profile of the dregs, on the contrary, shows less intense peaks compared to the essential oil which elute mainly between 30 and 35 min. Example 5: Highlighting the major phytochemical differences between an essential oil and the purified extract obtained according to example 1

[0180] The purified rosewood extract obtained according to Example 1 was compared to an essential oil of Aniba rosaeodora as prepared in Example 3 by high performance liquid chromatography (HPLC) analysis coupled with a diode array detector and an evaporative light scattering detector (ELSD) of solutions of the same concentration of these two extracts. The chromatographic analysis was carried out on a Core-Shell C18 column according to an elution gradient using mobile phases acidified with formic acid (HCO2H) consisting of a water / acetonitrile (ACN) / Propane-2-ol (IPA) mixture for route A and ACN / IPA / Methanol (MeOH) for route B.

[0181] Figure 2 represents the UV chromatographic profile at 300 nm of a rosewood essential oil as described in Example 3 and of the purified extract of exhausted rosewood according to Example 1, analyzed by HPLC / UV-ELSD under the following elution gradient conditions: 0-5 min 100% A, 5-22 min change from 100% A to 100% B then 22-32 min 100% B with 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 detector response in mAU is shown on the y-axis. The abscissa represents the analysis time in minutes.

[0182] As shown in Figure 2, the majority of semi- to non-volatile compounds corresponding to sesquiterpene derivatives and aryl phenyl ketones are not or weakly detected in rosewood essential oil. ISP‐BV23‐200PCT Description

[0183] In parallel, the two extracts obtained according to examples 1 and 3 were also compared by GC / MS-FID analysis. The samples were analyzed at the same headspace (HS) concentration. The separation was obtained on a 30 x 0.25 m, 0.25 µm apolar column, with elution according to a temperature gradient (isotherm 50°C, 5 min; then 3°C / min up to 120°C; then 5°C / min up to 250°C; isotherm 250°C, 5 min). The carrier gas used is helium.

[0184] Figure 3 represents the GC / MS chromatographic profiles of a rosewood essential oil as described in Example 3 and of the purified rosewood extract obtained according to Example 1, analyzed by GC / MS-FID under the conditions described above. The detector response is shown on the y-axis. The abscissa represents the analysis time in minutes.

[0185] Figure 3 illustrates the absence or presence in very small quantity of volatile compounds in the solubilized extract of exhausted rosewood compared to the essential oil diluted to 0.5% in glycerol tricaprylate / caprate which contains, as described in the prior art, a significant quantity of linalool (majority peak between 18 and 19 min) whose content is estimated between 70 and 90%. The chromatographic profile of the purified rosewood extract shows much lower peak abundances than the rosewood essential oil, with a linalool content estimated between 0.05.10-3 and 0.005%. This clear decrease in volatile compounds, particularly linalool, is the consequence of the steam distillation process previously applied to the rosewood which has exhausted the raw material in volatile compounds.

[0186] Figure 4 shows the HPLC / DEDL chromatographic profiles of the 1% crude rosewood extract in ethanol compared to the 1% purified rosewood extract in glycerol tricaprylate / caprate, obtained according to Example 1. The chromatographic analysis was carried out on a Core-Shell C18 column according to an elution gradient using mobile phases acidified with formic acid (HCO2H) consisting of a mixture of water / acetonitrile (ACN) / Propan-2-ol (IPA) for route A and ACN / IPA / Methanol (MeOH) for route B under the following elution gradient conditions: 0-5 min 100% A, 5-22 min transition from 100% A to 100% B then 22-32 min 100% B with 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 detector response in mAU is shown on the y-axis. The abscissa represents the analysis time in minutes. ISP‐BV23‐200PCT Description

[0187] A clear decrease in the intensity of the anibine peak (retention time at 9 min) is observed for the purified extract, unlike the crude extract for which anibine is one of the major compounds. These profiles are illustrated in Figure 4 and demonstrate the depletion of anibine in the purified extract, by precipitation of this compound following dilution in the carrier solvent. Example 6: Evaluation of the purified 1% rosewood extract on the expression of type I and type III collagen and hyaluronic acid on human skin biopsies

[0188] The aim of this experiment is to demonstrate an effect of purified rosewood extract on the synthesis of collagen I, collagen III, and hyaluronic acid in ex vivo human skin biopsies.

[0189] Protocol:

[0190] The expression of collagen I, collagen III, and hyaluronic acid was evaluated by indirect immunofluorescence on skin biopsies, previously treated by topical application of the purified rosewood extract obtained according to Example 1 and diluted to 1% in glycerol tricaprylate / caprate for 48 hours (twice a day). Control biopsies incubated in parallel under the same conditions received the placebo (Phosphate Buffer Saline, PBS). At the end of the incubation, the biopsies were fixed and embedded in paraffin for histological sections. Detections of collagen I, collagen III, and hyaluronic acid were performed by incubation with anti-Collagen I antibody (Abcam); anti-Collagen III (Proteintech), and a biotinylated HABP (hyaluronan-binding protein) probe (millipore).After 1.5 hours of incubation followed by rinsing, the sections were incubated in the presence of the anti-rabbit secondary antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen) to detect collagens I and III. A biotin / streptavidin system coupled to a fluorophore (Alexa Fluor® 488, Invitrogen) was used to detect hyaluronic acid. The sections were then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). The expression of collagen I, collagen III, and hyaluronic acid were then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0191] Results: As shown in Figure 5, when biopsies were treated with 1% purified rosewood extract, collagen I expression increased by 20%, ISP‐BV23‐200PCT Description Collagen III expression increased by 32%, and hyaluronic acid expression increased by 36% compared to the negative control.

[0192] Conclusion: Purified rosewood extract showed a positive effect on the expression of collagen I, collagen III, and hyaluronic acid. Example 7: Evaluation of 1% purified rosewood extract on the expression of Fibulin-7 in human skin biopsies

[0193] Purified rosewood extract was tested on the expression of

[0194] Principle: The aim of this experiment is to demonstrate an effect of purified rosewood extract on the synthesis of fibulin-7, in ex vivo human skin biopsies. Fibulin-7 is a protein preserving stem keratinocytes and identified as involved in facial morphology.

[0195] Protocol: Fibulin-7 expression is assessed by indirect immunofluorescence on skin biopsies, processed as in Example 6. Fibulin-7 detection is performed by incubation with anti-fibulin-7 antibody (Thermo Scientific). After 1.5 hours of incubation followed by rinsing, the sections are incubated in the presence of the anti-rabbit secondary antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Fibulin-7 expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0196] Results: When biopsies were treated with 1% purified rosewood extract, fibulin-7 expression increased by 39% compared to the negative control (mean + / - SEM, n=6 highly significant result with Student's t test).

[0197] Conclusion: Purified rosewood extract showed a positive effect on fibulin-7 expression. Example 8: Evaluation of 1% purified rosewood extract on collagen XVII expression in human skin biopsies

[0198] Purified rosewood extract was tested on the expression of collagen XVII, a protein whose expression decreases with age in human skin (Xiang Y, et al., J Cell Commun Signal.2022;16(3):421-432.), and identified as involved in the facial morphology of Europeans (Liu F. et al., PLoS Genet.2012;8(9):e1002932). ISP‐BV23‐200PCT Description

[0199] Protocol: Collagen XVII expression is assessed by indirect immunofluorescence on skin biopsies, processed as in Example 6. Collagen XVII detection is performed by incubation with anti-Col17A1 antibody (Abcam). After 1.5 hours of incubation followed by rinsing, the sections are incubated in the presence of the anti-rabbit secondary antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Collagen XVII expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0200] Results: When biopsies were treated with the purified rosewood extract according to Example 1 at 1%, the expression of collagen XVII increased by 94% compared to the negative control (mean + / - SEM, n=6 highly significant result with Student's test).

[0201] Conclusion: Purified rosewood extract showed a positive effect on the expression of collagen XVII. Example 9: Evaluation of 1% purified rosewood extract on harmonin expression in human skin biopsies

[0202] Purified rosewood extract was tested on the expression of harmonin, a protein that has scaffolding and molecular hub properties in signaling pathways (Bugge K, et al., J Biol Chem.2021;296:100226; Friis Theisen F., et al., J Biol Chem.2022;298(6):101963).

[0203] Protocol: Harmonin expression is assessed by indirect immunofluorescence on skin biopsies, treated as in Example 6. Harmonin detection is performed by incubation with anti-harmonin antibody (Proteintech). After one and a half hours of incubation followed by rinsing, the sections are incubated in the presence of the secondary anti-rabbit antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). Collagen XVII expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0204] Results: When biopsies were treated with 1% purified rosewood extract, harmonin expression increased by 44% compared to the negative control (mean + :- SEM, n=6 highly significant result with Student's test). ISP‐BV23‐200PCT Description

[0205] Conclusion: Purified rosewood extract showed a positive effect on harmonin expression. Example 10: Evaluation of 1% purified rosewood extract on CRABP2 expression in human skin biopsies

[0206] The purified rosewood extract was tested on the expression of CRABP2, a protein that decreases in human skin with age, and is involved in the retinoic acid signaling pathway.

[0207] Protocol: CRABP2 expression is assessed by indirect immunofluorescence on skin biopsies, treated as in Example 6. Biopsies serving as positive controls are incubated in parallel under the same conditions and receive 1 µg / ml and 10 µg / ml of retinoic acid (Sigma). CRABP2 detection is performed by incubation with anti-CRABP2 antibody (Abcam). After one and a half hours of incubation followed by rinsing, the sections are incubated in the presence of the anti-rabbit secondary antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). CRABP2 expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0208] Results: As shown in Figure 6, when biopsies were treated with the purified rosewood extract according to Example 1 at 1%, CRABP2 expression increased by 80%. When biopsies were treated with 1 µg / ml and 10 µg / ml of retinoic acid, CRABP2 expression increased by 106% and 132%, respectively.

[0209] Conclusion: Purified rosewood extract showed a positive effect on CRABP2 expression. Example 11: Evaluation of 1% purified rosewood extract on the expression of type V collagen alpha 3 chain in human skin biopsies

[0210] Purified rosewood extract was tested on the expression of the alpha 3 chain of type V collagen. Genetic variations of this gene have been associated with wrinkles located in the corner of the eye.

[0211] Protocol: The expression of the alpha 3 chain of type V collagen is evaluated by indirect immunofluorescence on skin biopsies, treated as in example 6. Biopsies serving as positive controls are incubated in parallel under the same conditions and receive 1 µg / ml of retinoic acid (Sigma). Detection of the chain ISP‐BV23‐200PCT Description Alpha 3 of type V collagen is performed by incubation with the alpha 3 chain antibody of type V collagen (Sigma). After 1.5 hours of incubation followed by rinsing, the sections are incubated in the presence of the secondary anti-rabbit antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). The expression of the alpha 3 chain of type V collagen is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0212] Results: As shown in Figure 7, when biopsies were treated with 1% purified rosewood extract, the expression of type V collagen alpha 3 chain increased by 23%. When biopsies were treated with 1 µg / ml retinoic acid, the expression of type V collagen alpha 3 chain increased by 28%.

[0213] Conclusion: Purified rosewood extract showed a positive effect on the expression of the alpha 3 chain of type V collagen. This anti-aging effect is particularly interesting for Asian skin. Example 12: In vivo (clinical) evaluation of purified rosewood extract on facial sagging, and signs of aging such as volume changes and the appearance of wrinkles

[0214] Principle: Study the in vivo efficacy of the purified rosewood extract according to example 1, formulated at 1% and make the link with the in vitro results obtained on key markers of facial morphology.

[0215] Protocol: Double-blind comparative study against a placebo, conducted on the faces of 33 volunteers (aged 35 to 67 years) randomly divided into two groups: 16 volunteers for the placebo group and 17 volunteers for the treated group, homogeneous according to age and sex.

[0216] [Table 4]: Skincare formula containing the rosewood extract of Example 1 at 1% used in the clinical test on the treated group. ingredients (trade name | INCI) % w / w Supplier ISP‐BV23‐200PCT Description Acid Copolymer (and) extract.

[0218] Duration of study: 56 days.

[0219] Follow-up visits on D0 (first day of the study), D28 and D56. ISP‐BV23‐200PCT Description

[0220] –Representative measurements of facial sagging: the angle of the V-shape of the face, the distance between the ptoses, the volume of the ptoses, the volume of the oval of the face as well as the volume of the cheekbones by the AEAVA-HE® system (Eotech®).

[0221] – Measurement of lip corner volume and roughness using the AEAVA-HE® system (Eotech®).

[0222] - Topography of the crow's feet skin by silicone replica, analyzed by fringe projection and stereometry associated with the AEAVA-HE® system (Eotech®). The profile roughness of the crow's feet wrinkles was measured and 2D and 3D representations of the skin topography were made.

[0223] - Color photos of the face with the HeadScan* V03 bench (Orion Concept*).

[0224] - Evaluation on a scale of 0 to 100 by an expert and by volunteers of the condition of the facial skin to the touch and according to the visual aspect of the following parameters:

[0225] – Skin tone: 0 corresponds to very dull skin and 100 to perfect luminosity and complexion.

[0226] – Crow's feet: 0 corresponds to smooth skin without fine lines and 100 to skin with marked microrelief and visible fine lines.

[0227] – nasolabial fold: 0 corresponding to a very deep nasolabial fold and 100 corresponding to a nasolabial fold that is not marked and not visible

[0228] - Skin tone: 0 corresponding to loose skin without tone and 100 corresponding to very firm skin with very good tone

[0229] - Oval facial contour: 0 corresponding to a poorly defined facial contour with pronounced ptosis and 100 corresponding to a perfectly oval facial contour without ptosis

[0230] - Facial harmony: 0 corresponding to an unharmonious face, a dull complexion, deep wrinkles and a nasolabial fold present, sagging skin and the oval of the face not at all defined with the presence of ptosis. 100 corresponding to a harmonious face with a radiant complexion, smooth skin without wrinkles, toned skin and the oval of the face perfectly defined.

[0231] The measurements were taken on the subject after 15 minutes of rest in a room at 21°C + / -1 and with a relative humidity of 50% + / - 5. ISP‐BV23‐200PCT Description

[0232] The results of the facial sagging and volume change measurements show that after one and two months of application, the volunteers in the treated group have a more V-shaped facial angle and a reduced volume of the facial oval, unlike the volunteers in the placebo group. These results are illustrated in Figure 8. In addition, as illustrated in Figure 9, the distance between the ptoses as well as their volumes are reduced compared to the placebo group.

[0233] These results are in agreement with the results obtained in vitro on collagen XVII but also harmonin and fibulin-7, key markers in facial morphology and decreasing with age.

[0234] Results of lip corner volume and roughness measurements: As shown in Figure 10, after 2 months of application, a decrease in the volume and roughness of lip corner wrinkles is observed for the treated group compared to the placebo group.

[0235] These results are consistent with the results obtained in vitro on Fibulin-7 (a key marker of lip corner wrinkles and decreasing with age).

[0236] Results of skin topography measurements at the crow's feet: as illustrated in Figure 11, a reduction in the roughness of crow's feet wrinkles is observed at D28 and D56 for the treated group compared to the placebo group.

[0237] These results support the results obtained in vitro on collagen I and III.

[0238] These results are confirmed by the observation of color photos of the face and the evaluation of the expert and the volunteers.

[0239] Conclusion of the test: the above results demonstrate that the purified rosewood extract according to example 1 formulated at 1% has a firming effect, helps to reduce facial sagging and redraw the oval of the face and has an anti-wrinkle effect allowing the reduction of wrinkles at the corners of the lips and crow's feet.

[0240] Similar results were obtained in a clinical study conducted on Asian skin in which a reduction in wrinkles was observed after one month of application with the same cream containing the purified rosewood extract formulated at 1%. This result is consistent with the in vitro results obtained at the level of the alpha 3 chain of type V collagen, a marker associated with wrinkles at the corner of the eye mainly in Asian skin. ISP‐BV23‐200PCT Description Example 13: Evaluation of 1% purified rosewood extract on the expression of interleukin 1 receptor type 1 (IL1R1) in human skin biopsies

[0241] Purified rosewood extract was tested for the expression of IL1R1, a protein that increases in human skin with inflammation.

[0242] Protocol: IL1R1 expression is assessed by indirect immunofluorescence on skin biopsies, treated as in Example 6. Biopsies serving as positive controls are incubated in parallel under the same conditions and receive 1 µg / ml and 10 µg / ml of retinoic acid (Sigma). IL1R1 detection is performed by incubation with anti-IL1R1 antibody (Tebu bio). After one and a half hours of incubation followed by rinsing, the sections are incubated in the presence of the secondary anti-rabbit antibody coupled to a fluorophore (Alexa Fluor® 488, Invitrogen). The sections are then examined under an Epi-fluorescence microscope (Zeiss Axiovert 200M microscope). IL1R1 expression is then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0243] Results: As shown in Figure 12, when biopsies were treated with the purified rosewood extract according to Example 1 at 1%, IL1R1 expression decreased by 3%. When biopsies were treated with 1 µg / ml and 10 µg / ml of retinoic acid, IL1R1 expression increased by 19% and 56%, respectively.

[0244] Conclusion: Purified rosewood extract showed no significant modulation of interleukin 1 receptor type 1 (IL1R1) expression. Retinoic acid application showed an increase in IL1R1 expression.

Claims

BV23‐200PCT Claims Claims

1. A process for obtaining a rosewood extract obtained from leaves or shavings of trunks and branches of exhausted rosewood (Aniba rosaeodora) comprising the following steps: a) adding between 5 and 25% of water to the exhausted and dried rosewood; b) carrying out an extraction using carbon dioxide (CO2) in the supercritical state in the presence of a polar co-solvent chosen from primary or secondary alcohols, or any mixture thereof; c) adding a support solvent chosen from polyol type solvents, saturated or unsaturated fatty alcohols, linear or branched, comprising from 8 to 30 carbons, glyceride type solvents or long carbon chain ester type solvents, or any mixture thereof; d) the extract solubilized in step c) is evaporated to remove all of the co-solvent,e) the extract is filtered in order to remove the precipitated compounds which do not dissolve in the support solvent and to collect the filtrate, f) the extract obtained in e) is diluted in the same support solvent at a concentration of between 0.005 and 50% of crude extract by weight of the total weight of the final extract.

2. Process according to claim 1 in which in step b) the co-solvent is ethanol, advantageously at a concentration of between 80 and 100% (volume / volume percentage of water), preferably between 90 and 100% and even more preferably at a concentration of 96%.

3. Method of one of the preceding claims wherein in step b), the mass ratio of the co-solvent relative to the carbon dioxide (CO2) in the supercritical state is between 0.050 and 0.080, advantageously between 0.055 and 0.075 and preferentially between 0.060 and 0.

070.

4. The method of one of claims 2 or 3 wherein in step b) the flow rate of co-solvent is between 10 and 20 ml / min.

5. The method of one of the preceding claims wherein in step b) the extraction temperature is between 35 and 85°C, BV23‐200PCT Claims advantageously between 45 and 75°C and preferably between 55 and 65°C and the pressure within the extractor is between 90 and 1000 bar, preferably between 150 and 700 bar and even more preferably between 250 and 350 bar.

6. Process of one of the preceding claims wherein in step c) the support solvent is chosen from solvents of polyol type, saturated or unsaturated fatty alcohol, linear or branched, comprising from 8 to 30 carbons or solvents of glyceride type or of ester type with a long carbon chain, or any mixture thereof, preferably the support solvent is glycerol tricaprylate / caprate.

7. Purified extract of rosewood obtainable by the process of one of claims 1 to 6, characterized in that it is diluted to a concentration of between 0.1 and 1% in the carrier solvent and comprises from 0.3 to 0.5%, advantageously from 0.35 to 0.5% and preferably between 0.4 and 0.5% of semi- to non-volatile compounds.

8. Purified rosewood extract of claim 7, characterized in that the semi- to non-volatile compounds comprise from 0.01 to 0.4% sesquiterpene derivatives in cotoine equivalent, from 0.001 to 0.1% aryl phenyl ketones including 0.001 to 0.05% cotoine, from 0.001 to 0.01% beta-sitosterol and from 0.001 to 0.1% anibine.

9. Composition comprising an effective amount of purified rosewood extract of one of claims 7 or 8 as active agent and a physiologically acceptable medium.

10. Composition of claim 9, characterized in that it comprises between 0.01% and 10%, preferably between 0.1 and 5% and even more preferably between 0.5 and 2% of purified rosewood extract by weight relative to the total weight of the composition.

11. Cosmetic use of the composition of claim 9 for preventing or limiting the signs of skin aging, firming the skin, limiting. BV23‐200PCT Claims the appearance of facial ptosis, redraw the contour of the face and also to limit the appearance of wrinkles and fine lines.

12. Cosmetic use of the composition of claim 9 to increase the expression of collagen I, collagen III, collagen XVII, hyaluronic acid, fibulin-7, harmonin and CRABP2.

13. Cosmetic use of the composition of claim 9 to increase the expression of the alpha 3 chain of type V collagen and to limit the appearance of wrinkles and fine lines.

14. Cosmetic use of the composition of claim 9 to obtain an effect similar to that of retinoids on the signs of skin aging, without exhibiting undesirable inflammatory effects.

15. Composition according to claim 9 for its use in combating external attacks producing pro-inflammatory free radicals, such as stress or pollution.

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