Method for obtaining an aqueous extract of lavender, compositions containing such extracts, and cosmetic uses thereof

Abstract

The present invention relates to a method for obtaining an aqueous lavender extract that is free of DNA and enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds and organic acids, by contacting the aerial parts of lavender with water, adding phytic acid at a pH between 10 and 11, adjusting the pH of the resulting mixture to a value between 6 and 8, and purifying the resulting mixture.The present invention also relates to cosmetic compositions containing such extracts and their cosmetic uses for protecting the skin from external aggressions and oxidation, preventing the signs of skin aging, improving photoprotection, whitening the skin, improving moisture retention, strengthening the barrier function, soothing the skin or improving biological mechanisms related to nighttime skin repair.

Description

[Technical Field]

[0001] The present invention relates to the field of cosmetics, and in particular to active ingredients of natural origin that are used in the preparation of cosmetic formulations for improving the appearance of or protecting the skin.

[0002] The present invention relates to a method for obtaining an aqueous extract of lavender, an extract obtained by this method enriched in small RNAs, sugars, phenolic compounds and organic acids, a cosmetic composition containing such an extract, and its cosmetic use in skin care, scalp and appendage care, more particularly for protecting the skin from external aggressions and oxidation, preventing the signs of skin ageing, improving photoprotection, skin lightening, improving skin moisture retention, strengthening the barrier function, skin soothing or improving biological mechanisms related to night-time skin repair. [Background technology]

[0003] The genus Lavandula, commonly known as lavender, comprises a group of 47 species. The best-known and most widely used species are true lavender, Lavandula angustifolia, a wild species native to Provence (southeastern France), and lavandin, a hybrid resulting from a cross between true lavender and spike lavender.

[0004] True lavender is widely used in Europe, North Africa, the Middle East, and Asia for the production of essential oils, particularly for the fragrance industry.

[0005] Essential oils (EOs) are typically obtained by steam distillation: previously dried flowers are exposed to a steam stream to remove volatile or soluble components, which are then recondensed to yield a hydrolate (or floral water) and a supernatant liquid containing the lipid portion of the plant, which constitutes the essential oil.

[0006] Lavender EO is highly fragrant and primarily contains volatile monoterpenes, such as linalool and linalyl acetate. Authentic lavender EO contains approximately 30% of both. Lavender EO is known for its soothing, pain-relieving, analgesic, anti-inflammatory, antiseptic, and antibacterial properties, among others. It also possesses antispasmodic and decongestant properties, and is indicated for soothing skin conditions and promoting healing. Furthermore, lavender EO is indicated for the improvement of sleep disorders and various tension states (e.g., anxiety, stress).

[0007] Lavender floral water contains less than a few percent of the same volatile organic compounds and water-soluble plant compounds found in the essential oil. Floral water has similar properties to lavender essential oil, but is more diluted due to a lower concentration of terpene compounds.

[0008] The use of lavender essential oil and floral water in cosmetics is very limited, in part due to the presence of many terpene molecules known for their irritating effects on the skin: for example, linalool is recognized as a potential allergen, and the use of lavender essential oil is not recommended for pregnant women and children under the age of 12.

[0009] Most of the lavender extraction methods described in the prior art use non-polar organic solvents (CN10338583, JP11199469) or supercritical fluids (KR2015042999), which allow extraction of mainly volatile aroma compounds. In the latter case, the resulting extract is rich in polyphenols and flavonoids, but does not contain other compounds of interest, such as amino acids, organic acids, or proteins. Furthermore, phenolic acids are not extracted using this type of technique, since these molecules are mainly extracted with polar solvents, ideally water.

[0010] Consumers of cosmetic products desire formulations that are as natural as possible and that are as effective as or more effective than synthetic products.

[0011] Although a variety of anti-aging cosmetic products are available on the market, there is an ever-increasing demand for new, effective, naturally derived cosmetic ingredients.

[0012] One of the problems that the present invention aims to solve is to provide a novel aqueous lavender extract that meets the requirements of the current cosmetic market in terms of naturalness criteria and has excellent biological efficacy.

[0013] Another problem that the present invention aims to solve is to provide a novel aqueous lavender extract that does not have the drawbacks of known extracts, namely, the strong odor, the instability of EO in formulations for topical application, or the irritating or allergenic properties due to the presence of terpene molecules such as linalool.

[0014] Another problem that the present invention aims to solve is to provide a novel aqueous lavender extract that is enriched in compounds known to be beneficial to the skin, such as small RNAs, sugars, phenolic compounds, and organic acids.

[0015] The present inventors have discovered a novel lavender flower extract that is particularly concentrated in small RNAs, sugars, phenolic compounds, and organic acids, and that does not have the drawbacks of prior art methods, such as the use of potentially toxic detergents and solvents in cosmetics.

[0016] Such extracts can be used in cosmetics for skin care, scalp and skin appendage care, more particularly for protecting the skin from external aggressions and oxidation, combating the signs of skin ageing, improving photoprotection, whitening the skin, improving skin moisture retention, strengthening the barrier function or soothing the skin. Summary of the Invention

[0017] The present invention provides a first method for obtaining an aqueous extract of the aerial parts of lavender, comprising the steps of: a) contacting the aerial parts of lavender with water; b) adding phytic acid to the mixture obtained in a) at a concentration between 1 and 10 mM and at a pH between 10 and 11; c) then adjusting the pH of the mixture obtained in b) to a value between 6 and 8; d) purifying the mixture obtained in c) to remove residual solid plant matter to obtain a purified aqueous crude extract; e) checking the pH and readjusting it if necessary to a value between 6 and 8, preferably between 6 and 6.5; The present invention relates to a method comprising:

[0018] Secondly, the present invention relates to an aqueous extract of the aerial parts of lavender that is free of DNA and is enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds, and organic acids, the aqueous extract being obtained by the method according to any one of claims 1 to 5, wherein the extract contains, by weight relative to the total weight of the extract, 2 to 10 g / kg of sugars, 100 to 1,500 mg / kg of organic acids, 500 to 2,000 mg / kg of phenolic compounds, and 40 to 200 mg / kg of low-molecular-weight RNAs up to 150 nucleotides in length, and has a dry weight of 10 to 30 g / kg.

[0019] The present invention thirdly relates to a cosmetic composition comprising an effective amount of the extract according to claim 6 or 7 as an active ingredient and a physiologically acceptable medium.

[0020] The present invention fourthly relates to the cosmetic use of the composition according to the invention for skin care, for the care of the scalp and skin appendages, and more particularly for protecting the skin from external aggressions and oxidation, for preventing the signs of skin ageing, for improving photoprotection, for lightening the skin, for improving skin moisture retention, for strengthening the barrier function, for soothing the skin or for improving the biological mechanisms associated with night-time skin repair.

[0021] The invention and its advantages will be better understood from the following description and non-limiting embodiments with reference to the accompanying drawings. [Brief explanation of the drawings]

[0022] [Figure 1] 1 is a graph showing the characterization of organic acids by HPLC-MS analysis. [Figure 2] Graph showing the analysis of small RNA by Bioanalyser 2100. A: Lavender extract obtained by the method according to the invention. B: Conventional lavender extract. DETAILED DESCRIPTION OF THE INVENTION

[0023] [Definition] All terms used herein have their most commonly known meanings unless otherwise specified. For purposes of the present invention, they are defined as follows:

[0024] "Lavender" refers to all species of the genus Lavandula and their hybrids (such as lavandin).

[0025] "Aerial parts" refers to the stems and flowers of lavender. Seeds are included in "aerial parts" within the meaning of the present invention.

[0026] "Aerial parts" and "flowers" are used interchangeably and refer to the flowers and the thin stems that bear the flowers.

[0027] "Small RNA" or "low molecular weight RNA" or "small RNA up to 150 nucleotides in length" refers to non-coding RNA (ribonucleic acid) up to 150 nucleotides in length and small molecular weight, including any type of single-stranded and / or double-stranded small non-messenger RNA, such as microRNA, interfering RNA, introns, small nuclear RNA, or any RNA fragment. Electrophoretic analysis has shown that the small RNAs present in the lavender extract of the present invention have various molecular weights ranging from approximately 30 to 150 nucleotides.

[0028] "Organic acid" means alpha-hydroxy acids (or AHAs), ie, carboxylic acids derived from fruit or plant sugars, including, for example, glycolic acid, malic acid, citric acid, tartaric acid, succinic acid, and uronic acid.

[0029] "Phenol compounds" (or polyphenols) refer to plant-derived molecules that contain an aromatic ring with one or more hydroxyl groups, such as phenolic acids, flavonoids, or their derivatives. Polyphenol compounds are known to be powerful antioxidant molecules.

[0030] "Sugars" means monosaccharides, especially glucose and fructose, as well as oligosaccharides and polysaccharides.

[0031] "Phytomolecules of interest" means any molecule present in the lavender extract according to the invention, in particular small RNAs up to 150 nucleotides in length, sugars, phenolic compounds and organic acids.

[0032] Where a range of numerical values ​​is stated, the limits of that range should be understood to expressly include all intervening values ​​within the range. For example, a numerical range between 1% and 10% should be understood to include 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, and 10%, as well as all fractional values ​​between 1% and 10%.

[0033] Numerical percentages are percentages by weight, ie weight of compound relative to the total weight of the intended mixture, unless otherwise specified.

[0034] The compositions described herein may "comprise," "consist of," or "consist substantially of" essential compounds or optional ingredients.

[0035] "Consisting essentially of" means that the composition or component may contain additional ingredients, but only if the additional ingredients do not alter the basic or novel properties of the composition or its uses as described herein.

[0036] "Physiologically acceptable medium" means an excipient that is suitable for contact with the outer layer of the skin or mucous membranes and that does not cause toxicity, irritation, excessive allergic or intolerance reactions, etc., and that is commensurate with a reasonable benefit / risk ratio.

[0037] "Topical application" means applying or spreading the aqueous extract of the present invention, or a composition containing the same, enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds and organic acids, onto the surface of the skin or mucous membrane.

[0038] "Skin" refers to the skin of the face, especially around the eyes and mouth, nose, forehead, neck and hands, as well as the skin of the entire body.

[0039] "Scalp" means the skin overlying the skull, including hair follicles and inter-follicular skin spaces.

[0040] "Skin appendages" refers to the keratin-rich products of the hair follicle (hair and body hair) and nails.

[0041] "Enhancing the barrier function" means improving the protective properties of the skin against external aggressions (UV radiation, visible or infrared light, environmental pollutants, microorganisms, etc.).

[0042] "Skin soothing" means reducing irritation reactions that manifest themselves as discomfort such as redness.

[0043] "Skin lightening" means reducing the intensity of skin color related to the melanin content of the epidermis either homogeneously or by local means by acting on pigmentary disorders such as age spots or senile lentigines.

[0044] "Effective amount" means the minimum amount of the extract according to the invention that is necessary to obtain at least one of the biological activities, in particular exhibiting antioxidant activity against reactive oxygen species, increasing melatonin, or attempting to decrease melanin, or any other biological marker tested, without this amount being toxic.

[0045] "Skin moisture retention" refers to the presence and distribution of moisture in the upper layer of the epidermis.

[0046] "Improved skin hydration" refers to improving the changes in the appearance of the skin due to dehydration, such as dryness, tightness, and discomfort, whether this condition is related to internal factors or external factors such as adverse environmental conditions.

[0047] "Signs of skin aging" means any change in the appearance of the skin due to aging, e.g., wrinkles and fine lines, cracks, bags under the eyes, dark circles, atrophy, loss of skin elasticity, firmness and / or tone, as well as any internal changes in the skin that do not systematically result in a change in appearance, e.g., thinning of the skin, or any internal deterioration of the skin resulting from environmental stresses such as environmental pollution and solar radiation, including UV radiation.

[0048] "Signs of skin aging" include pigmentary disorders such as senile or solar lentigines.

[0049] "External attacks" refers to environmental pollution originating from the ambient atmosphere inside and outside the home, including solar radiation, including visible light, UV radiation and infrared radiation, particles of various sizes (PM10 of 10 μm, PM2.5 of 2.5 μm or ultrafine particles less than 100 nm), and some chemical elements (volatile organic compounds, polycyclic aromatic hydrocarbons, heavy metals, etc.).

[0050] "Improved skin appearance" means that the skin appears finer, more radiant, and has an even skin tone.

[0051] It is to be understood that the present invention relates to mammals, and more particularly to humans.

[0052] [Extraction method] Classical ribonucleic acid (RNA) extraction protocols use solvents that are not suitable for cosmetic use (Zumbo, p. 2014 "Phenol-chloroform Extraction", 2014). These methods aim to obtain completely purified nucleic acids (RNA, DNA, or small RNA), i.e., free from secondary metabolites, vitamins, sugars, peptides, and any other molecules of interest that have beneficial effects on the skin and are of cosmetic interest.

[0053] FR2831168 also describes a method for obtaining plant extracts rich in nucleic acids (DNA and / or RNA), which uses cellulolytic enzymes.

[0054] Furthermore, patent documents EP1723958 and WO03101376 describe compositions for topical application that contain synthetic double-stranded RNA oligonucleotides of known sequence and 12 to 40 nucleotides in length that have siRNA (short interfering) function.

[0055] Furthermore, document FR1502361 (also published as WO2017084958) describes a method for obtaining an aqueous plant extract enriched in low molecular weight ribonucleic acid (RNA) for preparing a cosmetic composition, using EDTA at a concentration between 2 and 15 mM.

[0056] The advantage of using phytic acid instead of EDTA is that, unlike EDTA, phytic acid is a natural molecule found in the seed shells of grains and legumes. Using phytic acid makes it possible to obtain a 100% natural lavender extract while maintaining good extraction efficiency for plant molecules contained in the plant. The same extraction efficiency for small RNA also applies to other compounds present in lavender flowers, such as sugars, phenolic compounds, or organic acids, such as tartaric acid, malic acid, or citric acid.

[0057] The present invention relates firstly to the extraction method used to obtain an aqueous extract of dried or fresh aerial parts of lavender.

[0058] The extraction method of the present invention makes it possible to avoid the use of solvents that are not considered cosmetic solvents and to obtain extracts that are rich in plant molecules of cosmetic interest, such as small RNAs up to 150 nucleotides in length, sugars, phenolic compounds, and organic acids.

[0059] The method according to the invention reduces the impact on the environment.

[0060] In the first step a) of the method according to the present invention, the aerial parts of lavender are mixed with water. The water used can be distilled water, demineralized water, or water enriched with inorganic salts and / or trace elements. The water used is preferably distilled water.

[0061] Preferably, the above-ground parts of lavender are lavender flowers and the small stems on which the flowers grow.

[0062] Preferably, the lavender species used is Lavandula angustifolia, i.e. true lavender.

[0063] Preferably, the lavender aerial parts are in dry form.

[0064] Preferably, the aerial parts of lavender are ground to powder before being brought into the presence of water in step a). The grinding of the aerial parts of lavender results in a mechanical action that allows for better extraction. After mechanical grinding to obtain plant material in powder form, alkaline dissolution in the presence of phytic acid promotes the complete destruction of cell membranes, and in particular nuclear membranes. Preferably, the pre-ground aerial parts of lavender in powder form are mixed with water in step a) at a plant material / water ratio of 3 to 20% w / w, more preferably between 3 and 10%, for example 3%, 5% or 10% (w / w).

[0065] Next, in step b), phytic acid is added to the mixture obtained in step a). The pH in this step must be basic, between 10 and 11, and should be adjusted, if necessary, by adding sodium hydroxide (NaOH). A basic pH between 10 and 11 is essential in step b). Preferably, the pH is adjusted to a value between 10.5 and 11. In fact, this pH level, in conjunction with the action of phytic acid, causes the destruction of cell membranes, including the nuclear membrane, cell lysis, and DNA denaturation (separation of the two strands of the double helix). Phytic acid weakens and disrupts the pectocellulose membrane of plant cells by complexing and sequestering divalent ions, such as calcium ions, which form ionic bridges between pectin molecules surrounding cellulose microfibrils. This results in the enhanced release of cell contents during this extraction method. The phytic acid treatment step is essential for concentrating low-molecular-weight RNA in the extract and, more generally, for ensuring good extraction yields of other plant molecules of interest, namely sugars, phenolic compounds, and organic acids.

[0066] The pH is monitored and the mixture remains basic, stabilizing between 9 and 11 at the end of step b).

[0067] In such extraction methods, the use of an aqueous extraction solution containing a natural chelating agent such as phytic acid allows for the enrichment of low molecular weight RNA in the final extract. Phytic acid is a molecule that naturally occurs in the shells of seeds such as cereals and legumes. Phytic acid exists as a calcium salt, or more often a magnesium salt, and plays important roles in plants, for example, as a major source of phosphorus.

[0068] Preferably, the phytic acid used is phytic acid powder in the form of its sodium salt, preferably at a concentration between 1 and 10 mM, preferably between 1 and 5 mM, more preferably at a concentration of 3 mM.

[0069] As shown in Table 1 below, the present invention works particularly well when the phytic acid concentration is between 2 and 3. It can be seen that as little as 2.25 mM phytic acid produces the same results as 10 mM EDTA in terms of small RNA concentration, and similar overall extraction yields. A concentration of 3 mM also provides optimal extraction efficiency for small RNA. Furthermore, a 3 mM concentration is optimal for higher yields of other target compounds, such as sugars, phenolic compounds, and organic acids. At a phytic acid concentration of 4.5 mM, the small RNA extraction yield is even higher, while the overall extraction yield is lower.

[0070] [Table 1]

[0071] Step b) of the treatment with phytic acid preferably lasts for at least 1 hour at a temperature between 20 and 80° C. During this step, the mixture obtained in a) is advantageously stirred.

[0072] Advantageously, diatomaceous earth is added after step b) to allow easy separation of the solid residual plant material from the extract (soluble fraction) in the next step.

[0073] Then, in step c), the pH of the mixture obtained in b) is adjusted to a value between 6 and 8.

[0074] The pH can be adjusted by adding hydrochloric acid (HCl) solution or any other equivalent acid compatible with cosmetic use. Acidification causes DNA to rapidly renature (reunion of the double helix strands). However, chromosomal DNA is so long that it cannot completely reunite and forms insoluble tangles. Meanwhile, small RNAs remain in solution. In this way, DNA and small RNAs are separated into two distinct phases: a solid phase containing primarily chromosomal DNA and a liquid phase containing primarily small RNAs. The pH adjustment step in step d) of the method of the present invention is essential for optimal extraction of not only small RNAs but also other plant molecules of interest, namely sugars, phenolic compounds, and organic acids.

[0075] In step d), the mixture obtained in c) is purified to remove the residual solid aerial parts of lavender and recover the soluble portion constituting the aqueous crude extract of the present invention. Any method known to those skilled in the art can be used. For example, the mixture obtained in c) can be filtered through a filter with a porosity of more than 30 μm, and the filtrate recovered. Preferably, the mixture obtained in c) is centrifuged at low speed, for example at 4000 g for at least 10 minutes, to settle the residual plant material in the pellet and recover the aqueous crude extract in the supernatant.

[0076] In step e), the pH is checked and readjusted to a value between 6 and 8. Preferably, the pH is readjusted to a value between 6 and 6.5, even more preferably to a value of 6.5. The pH is readjusted by adding hydrochloric acid (HCl) solution or any equivalent acid compatible with cosmetic use.

[0077] In fact, a pH below 6 may precipitate nucleic acids in general, which may lead to the precipitation of small RNAs up to 150 nucleotides in length. The step of adjusting the pH in step e) of the method according to the invention is essential to have an optimal stability of the small RNAs in the extract.

[0078] After step e), a concentrated crude extract is obtained.

[0079] Advantageously, the readjustment of the pH in step e) is carried out after at least one filtration of the crude aqueous extract obtained in d), preferably by successive filtrations, lowering the filtration threshold from 20-50 μm to a sterile filtration of 0.1-0.3 μm.

[0080] The extract obtained in step e) can then be diluted with a physiologically acceptable solvent for cosmetic use so that its dry weight is between 4 and 20 g / kg of dry extract relative to the total weight of the diluted extract, this step improving the stability of the extract over time.

[0081] Secondly, the present invention relates to an aqueous extract of the aerial parts of lavender obtained by the above-mentioned method, which is enriched with small RNAs up to 150 nucleotides in length, sugars, phenolic compounds, and organic acids, and is free of DNA (deoxyribonucleic acid).

[0082] The present invention also relates to an aqueous extract of the aerial parts of lavender, obtained directly by the above method, enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds, and organic acids, which extract does not contain DNA (deoxyribonucleic acid).

[0083] Using the method of the present invention, steps a) through e) yield a concentrated aqueous crude extract of lavender with an amber to dark amber color. This extract contains 2-10 g / kg of sugars, 100-1500 mg / kg of organic acids, 500-2000 mg / kg of phenolic compounds, and 40-200 mg / kg of low molecular weight RNA up to 150 nucleotides in length, and has a dry weight of 10-30 g / kg. However, for the aerial parts, particularly lavender flowers of the species Lavandula angustifolia, the extract obtained can exhibit significant variability depending on factors such as harvest location, year, season, climatic conditions, and biotic stress.

[0084] Such extracts can then be diluted with a physiologically acceptable solvent for cosmetic use, and the concentration of the extract is then adjusted to between 4 and 20 g / kg dry weight of dry extract relative to the total weight of the diluted extract.

[0085] Illustrative, non-limiting examples of physiologically acceptable solvents include water, glycerol, ethanol, propanediol, and the natural version called Zemea® made from corn, butylene glycol, dipropylene glycol, ethoxylated or propoxylated diglycol, cyclic polyols, or any mixture of these solvents. The extract thus obtained can be diluted to obtain a final concentration of 50% plant-derived butylene glycol, 50% plant-derived propanediol, or 30% plant-derived glycerin.

[0086] Preferably, the extract obtained by the method according to the invention is diluted with butylene glycol so that the diluted extract contains a final butylene glycol concentration of 50%.

[0087] Such diluted extracts contain, by weight relative to the total weight of the extract, 4-20 g / kg of dry extract, 0.5-10 g / kg of sugars, 50-700 mg / kg of organic acids, 50-1500 mg / kg of phenolic compounds, and 10-100 mg / kg of low molecular weight RNA up to 150 nucleotides in length.

[0088] Non-limiting examples include a diluted extract of Lavandula angustifolia containing, among others, sugars at a concentration of 1.7 g / kg, organic acids at a content of 570 mg / kg, phenolic compounds at 620 mg / kg, and low molecular weight RNA up to 150 nucleotides in length at 45 mg / kg.

[0089] On the other hand, lavender floral water and essential oil contain mainly terpene scent molecules and do not contain low molecular weight RNA up to 150 nucleotides in length, sugars, phenolic compounds or organic acids.

[0090] Thus, the extracts of the present invention contain a wide range of plant molecules that may have beneficial effects on the skin without presenting the risk of skin irritation or other health hazards.

[0091] For example, sugars play an active role in hydrating the epidermal layers and in resisting external aggressions without exhibiting any undesirable effects. The lavender extract according to the invention contains in particular mono- and polysaccharides which are neither present in lavender floral water nor in the essential oil.

[0092] Lavender is a member of the Lamiaceae family, which has a unique metabolism known as Crassulacean Acid Metabolism (CAM). The plant stores organic acids, more specifically malic acid, citric acid, and tartaric acid, within its cells. The method of the present invention makes it possible to extract these organic acids or AHAs. When applied to the skin, these AHAs reduce cell cohesion between keratinocytes, causing desquamation of the stratum corneum and thereby promoting cell renewal.

[0093] The lavender extracts of the present invention are also enriched in phenolic compounds, such as phenolic acids. These water-soluble molecules known for their antioxidant activity contribute to the antioxidant and protective capabilities of the lavender extracts of the present invention.

[0094] A third aspect of the present invention is a cosmetic composition comprising as an active ingredient an effective amount of an aqueous extract obtained according to the present invention, which is enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds and organic acids, and a physiologically acceptable vehicle. The aqueous extract enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds and organic acids obtained according to the invention is advantageously used as an active ingredient for the preparation of a cosmetic composition.

[0095] Advantageously, the extract of the aerial parts of lavender according to the invention is added to the composition at a concentration ranging from 0.05 to 5% by weight relative to the total weight of the composition, preferably at a concentration ranging from 0.1 to 2.5% by weight relative to the total weight of the composition, and more preferably at a concentration ranging from 0.1 to 1.0% by weight relative to the total weight of the composition. The compositions usable in the present invention may be applied by any suitable route, in particular orally, or topically, and the formulation of the compositions will be adapted by those skilled in the art.

[0096] Preferably, the compositions according to the invention are in a form suitable for topical application, and therefore must contain a physiologically acceptable medium, i.e., be compatible with the skin and skin appendages, not risk discomfort during their application, and include any suitable cosmetic form.

[0097] The compositions for implementing the invention may in particular be in the form of an aqueous, hydroalcoholic or oily solution, an oil-in-water emulsion, a water-in-oil emulsion or multiple emulsions, and may also be in the form of a suspension or powder suitable for application to the skin, mucous membranes, lips and / or hair.

[0098] These compositions may be highly or poorly fluid and may have the appearance of a cream, lotion, milk, serum, ointment, gel, paste or foam. They may also be in solid form, such as a stick, or may be applied to the skin in the form of an aerosol. Examples of physiologically acceptable vehicles commonly used in the envisaged fields of application include adjuvants necessary for the formulation, such as solvents, thickeners, diluents, antioxidants, colorants, sun filters, self-tanning agents, pigments, fillers, preservatives, fragrances, odor absorbers, essential oils, vitamins, essential fatty acids, surfactants, film-forming polymers, etc.

[0099] In all cases, a person skilled in the art will select these adjuvants and their proportions in such a way as to ensure that the advantageous properties desired for the composition of the present invention are not impaired. These adjuvants can represent, for example, 0.01 to 20% by weight of the total composition. If the composition of the present invention is an emulsion, the fatty phase can represent 5 to 80% by weight, preferably 5 to 50% by weight, of the total weight of the composition. The emulsifiers and coemulsifiers used in the composition are selected from those conventionally used in the field under consideration. They can be used, for example, in proportions ranging from 0.3 to 30% by weight of the total weight of the composition.

[0100] According to another advantageous embodiment of the invention, the aqueous lavender extract according to the invention can be encapsulated or contained in a cosmetic vector, such as a liposome or any other nano- or microcapsule used in the cosmetics field, or adsorbed onto powdered organic polymers, mineral carriers such as talc and bentonite.

[0101] Advantageously, the composition according to the invention may contain, in addition to the active ingredient according to the invention, at least one other active agent having a cosmetic effect similar and / or complementary to that of said active ingredient.

[0102] For example, the additional active agent can be selected from anti-aging agents, stabilizers, whitening agents, moisturizing agents, draining agents, microcirculation promoters, exfoliating agents, exfoliating agents, extracellular matrix stimulators, energy metabolism activators, antibacterial agents, antifungal agents, soothing agents, anti-free radical agents, anti-UV agents, anti-acne agents, anti-inflammatory agents, anesthetic agents, warming agents, cooling agents, and slimming agents.

[0103] Such additional active agents include - Vitamin A, especially retinoic acid, retinol, retinol propionate, retinol palmitate, - Vitamin B3, especially niacinamide, tocopherol nicotinate, - Vitamin B5, Vitamin B6, Vitamin B12, Panthenol, - Vitamin C, especially ascorbic acid, ascorbyl glucoside, ascorbyl tetrapalmitate, magnesium, and sodium ascorbyl phosphate, - Vitamins E, F, H, K, PP, Coenzyme Q10, - metalloproteinase inhibitors or TIMP activators, - DHEA, its precursors and derivatives, - amino acids, such as arginine, ornithine, hydroxyproline, hydroxyproline dipalmitate, palmitoylglycine, hydroxylysine, methionine, and derivatives thereof, N-acylated amino acid compounds; - natural or synthetic peptides, including di-, tri-, tetra-, penta-, and hexapeptides and other species, such as their lipophilic, isomeric, complexed derivatives with metal ions (e.g., copper, zinc, manganese, magnesium, etc.), e.g., MATRIXYL®, ARGIRELINE®, CHRONOGEN™, LAMINIXYL IS™, PEPTIDE Q10™, COLLAXYL™ (Patent FR2827170, ASHLAND®), PEPTIDE VINCI 01™ (Patent FR2837098, ASHLAND®), PEPTIDE VINCI 02™ (Patent FR2841781, ASHLAND®), the peptide commercially known as ATPeptide™ (Patent FR2846883, ASHLAND®), or the synthetic peptide of the sequence Arg-Gly-Ser-NH2 sold by ASHLAND® under the name ATPeptide™; - Artemia salina extract sold under the name GP4G™ (FR2817748, ASHLAND®), plant peptide extracts, such as flax extract (Lipigenin™, patent FR2956818, ASHLAND®), extracts of soybean, spelt, grape, rapeseed, flax, rice, corn, pea, yeast extracts, such as Dynagen™ (patent FR2951946, ASHLAND™) or Actopontine™ (patent FR2944526, ASHLAND™), - Dehydroacetic acid (DHA), - phistosterols of synthetic or natural origin, - salicylic acid and its derivatives, alpha- and beta-hydroxy acids, silanols, - Amino sugars, glucosamine, D-glucosamine, N-acetylglucosamine, N-acetyl-D-glucosamine, mannosamine, N-acetylmannosamine, galactosamine, N-acetylgalactosamine, - Extracts of polyphenols, isoflavones and flavonoids, such as grape extract, pine extract, olive extract, lipids, such as ceramides or phospholipids, oils of animal origin, such as squalene or squalane, vegetable oils, such as sweet almond oil, copra oil, castor oil, jojoba oil, olive oil, rapeseed oil, peanut oil, sunflower oil, wheat germ oil, corn germ oil, soybean oil, cottonseed oil, alfalfa oil, poppy seed oil, pumpkin oil, evening primrose oil, millet oil, barley oil, rye oil, safflower oil, passion flower oil, hazelnut oil, palm oil, apricot kernel oil, avocado oil, calendula oil, ethoxylated vegetable oils, shea butter, - All UV screens and sun filters, - Cyclic AMP and its derivatives, adenylate cyclase enzyme activators and phosphodiesterase enzyme inhibitors, Centella asiatica extract, asiaticoside and asiatic acid, methylxanthines, theine, caffeine and its derivatives, theophylline, theobromine, forskolin, esculin and esculoside, ACE inhibitors, Val-Trp peptides, neuropeptide Y inhibitors, enkephalin, ginkgo biloba extract, dioscorea extract, rutin, yerba mate extract, guarana extract, oligosaccharides, polysaccharides, carnitine, ivy extract, rockweed extract, hydrolyzed pitcher plant extract, hydrolyzed cockscomb extract, Anogeissus leiocarpus extract, cassava extract utilissima leaf extract, palmitoylcarnitine, carnosine, taurine, elderberry extract, and seaweed extracts such as Palmaria palmata extract, It can be selected from the group consisting of:

[0104] Fourthly, the present invention relates to the cosmetic use of a composition comprising the lavender extract according to the present invention for skin care, for the care of the scalp and skin appendages, and more particularly for protecting the skin from external aggressions and oxidation, for preventing the signs of skin ageing, for improving photoprotection, for lightening the skin, for improving skin moisture retention, for strengthening the skin's barrier function, for soothing the skin, or for improving the biological mechanisms associated with night-time skin repair.

[0105] The skin is an organ composed of several layers (dermis, epidermis and stratum corneum) that covers the entire surface of the body and ensures protective functions against external aggression, sensory, immune, metabolic or thermoregulatory functions, or a barrier function that limits dehydration.

[0106] In particular, the stratum corneum functions as a protective physical barrier, commonly referred to as the "skin barrier function." This function is primarily important for tissue homeostasis and protection from the external environment.

[0107] The appearance of the skin can change due to internal changes (intrinsic aging, disease, and hormonal changes such as pregnancy) or external factors (environmental factors such as pollution, sunlight, pathogens, and temperature changes), all of which affect not only the skin but also its keratinous appendages, such as body hair, eyelashes, eyebrows, nails, and scalp hair.

[0108] The extract of the aerial parts of lavender according to the present invention has been tested for key biological markers related to nighttime skin repair mechanisms. Mechanisms occurring in the skin at night include increased DNA repair, increased cell proliferation rate, increased skin temperature, increased skin blood flow, increased incidence of itching, and increased water loss due to increased permeability of the skin barrier (Matsui M. Set. et al., Biological rhythms in the skin, Int. J. Mol. Sci. 2016, 17, 801). The extract of the aerial parts of lavender according to the present invention has been evaluated, in particular, for the repair rate of pyrimidine dimers (CPD stands for Cyclobutane Pyrimidine Dimers). The day / night rhythm, as detected at the central nervous system level, is the production of melatonin by the pineal gland (Slominski AT et al., Melatonin: A cutaneous perspective on its production, metabolism, and functions. J Invest Dermatol, 2018 March, 138(3):490-499). Melatonin is produced locally in the skin from tryptophan. These functions help to reduce the levels of reactive oxygen species (ROS) and reactive nitrogen species (RNS), respectively. In fact, in addition to its direct role as a free radical scavenger, melatonin promotes the production of antioxidant enzymes, such as catalase and superoxide dismutase, which are involved in DNA repair. By optimizing mitochondrial function, melatonin also helps increase the level of ATP produced in cells. The effect of the extract of the aerial parts of lavender according to the invention was assessed on melatonin levels in the skin and on the expression of the AANAT enzyme (aralkylamine N-acetyltransferase or serotonin N-acetyltransferase or timezyme), which catalyzes the N-acetylation of serotonin to N-acetylserotonin, the final step in melatonin synthesis.Additionally, topical application of exogenous melatonin has been shown to be effective in treating hair loss associated with androgenetic alopecia (Fisher TW, Topical melatonin for treatment of androgenetic alopecia, Int J Trichology, 2012 Oct-Dec, 4(4):236-245).

[0109] The extract of the aerial parts of lavender according to the present invention has been shown to be effective in increasing melatonin production in cultured skin biopsies.

[0110] Alterations in barrier function occur due to external aggressions, such as UV light. The consequences are profound in the loss of cell cohesion and mechanical integrity. Specific enzymes involved in the final stages of keratinocyte differentiation play a key role in UV protection. Alterations in the expression and / or activity of these enzymes have important effects on skin barrier function and homeostasis.

[0111] The extract of the aerial parts of lavender according to the invention has proven effective in improving the skin's protection against external aggressions (UV radiation, environmental pollution, microorganisms, etc.) by strengthening the barrier function.

[0112] The extract of the aerial parts of lavender according to the present invention has been shown to be effective in reducing melanin levels in skin biopsies and inducing a skin lightening effect. [Example]

[0113] In the following, an exemplary embodiment of the method according to the invention will be described.

[0114] [Example 1] <Preparation of small RNA-enriched lavender (Lavandula angustifolia) extract>

[0115] (Extraction method) Lavender flowers from the species Lavandula angustifolia were pre-dried in a well-ventilated area protected from light. In the first step, 3% of the dried lavender flowers and flower stems were ground to a powder with a particle size ranging from 500 μm to 1 mm, preferably 800 μm, equivalent to 30 g of dried lavender flower powder in 968 g of distilled water. Phytic acid was then added at 2 g / L or 3 mM. The pH was adjusted to 10.8 to optimally enrich the extract with low-molecular-weight RNA.

[0116] The mixture was then heated with stirring at 80° C. for 1 hour. After this period of extraction, diatomaceous earth was added to the mixture to facilitate subsequent separation of the extract (soluble fraction) from the solid residual plant material.

[0117] The mixture was then filtered through a 30 μm pore size filter to remove solids, and the pH was adjusted to 7.5 using HCl solution. To clarify the plant extract, successive filtrations were performed with decreasing filter porosity until a 0.2 μm sterile filtration was achieved.

[0118] After this step, the pH was checked and then adjusted to 6.3 with HCl solution. pH values ​​of 6 and 6.5 preserved small RNAs in the extract. An aqueous extract with a dry weight of 12.6 g / kg was obtained.

[0119] (Characterization of lavender extract) The resulting extract had a dry weight of 12.6 g / kg. Physicochemical analysis showed that the obtained extract had a concentration of 3.7 g / kg total sugars, 1160 g / kg total organic acids, 1270 mg / kg total phenolic compounds, and 118 mg / kg low molecular weight RNA of up to 150 nucleotides in length. The extract was then diluted with a physiologically acceptable cosmetic solvent, making it possible to ensure better stability and better preservation of the extract over time.

[0120] Dilution with plant-derived butylene glycol was performed to obtain a final concentration of 50% butylene glycol and 50% lavender extract. This diluted extract then had a dry weight of 6 g / kg, 1.7 g / kg total sugars, 570 mg / kg total organic acids, 620 mg / kg total phenolic compounds, and 45 mg / kg low molecular weight RNA up to 150 nucleotides in length.

[0121] (Assay methods used to measure the amount of various compounds in the final lavender extract) The total sugar content of the extracts was determined by spectrophotometric analysis based on an adaptation of the assay described by Dubois et al. (1956) (Dubois et al., "Colorimetric method for determination of sugars and related substances", Anal. Chem., 1956, 28(3), 350-356). This analysis consisted of dissolving the raw material in concentrated sulfuric acid and then reacting it with phenol to form a colored complex. The absorbance of the complex was read at 490 nm in a spectrophotometer. The sugar content was determined using a standard glucose curve. TLC analysis showed that the majority of the sugars present in the extracts according to the invention were glucose and fructose molecules, as well as higher molecular weight sugars (oligosaccharides and polysaccharides).

[0122] The total polyphenol content of lavender extracts was determined by Folin-Ciocalteu spectrophotometric analysis (Singleton et al., Analysis of total phenols and other oxidative and antioxidant substrates by means of the Folin-Ciocalteu reagent, 1999, 299:152). Polyphenolic compounds in the samples reacted with the Folin-Ciocalteu reagent, producing a blue color due to the oxidation of the reagent. The absorbance of the samples was read at 760 nm on a spectrophotometer. The content was expressed as gallic acid equivalents using a gallic acid standard curve.

[0123] Organic acid characterization was performed on the lavender extract, floral water, and reference essential oils from Example 1. High-performance liquid chromatography analysis was performed using a mass spectrometer. All samples were separated on an Agilent 1260 HPLC system (Agilent Technologies) using an EC 150 / 4.6 Nucleoshell RP 18plus-5 μm column (150 × 4.6 mm) (Macherey Nagel: 763236.46). The flow rate was 0.3 ml / min. The mobile phase consisted of 0.01% formic acid (HCOOH) solution (A) and acetonitrile (B). A gradient program facilitated elution as shown in Table 2.

[0124] [Table 2]

[0125] The column temperature was maintained at 25°C, and the injection volume was 5 μL. Detection was performed in negative mode on an ACQUITY Qda mass spectrometer detector (WATERS) equipped with an electrospray ion source. The electrospray ion source was set at a capillary voltage of 0.8 kV and a probe temperature of 600°C. The M / z and cone voltage were set to target each compound as shown in Table 3.

[0126] [Table 3]

[0127] Identification of organic acids was performed by comparing the retention time and mass spectral peaks of the samples with those of standards. Quantitative estimation of organic acids was based on comparison of the area maxima of the sample concentration with the area maxima of the standards.

[0128] HPLC-MS analysis, which can quantify and identify the organic acids contained in the extract, showed that only lavender extract contained various types of organic acids, mainly citric acid, malic acid, and tartaric acid, as shown in Figure 1. HPLC-MS analysis showed that these organic acids were not present in true lavender floral water or true lavender essential oil.

[0129] Quantification of small RNAs was performed using a miniaturized electrophoresis method (Bioanalyser 2100®, Agilent) on a microfluidic chip specialized for nucleic acid analysis, including small RNAs. This method allows the size and concentration of nucleic acids contained in extracts to be measured from a few microliters. The results are shown as graphs with arbitrary fluorescence units (FU) on the vertical axis and nucleotide numbers (nt) on the horizontal axis. An internal marker (the 25-nt peak in Figure 2) was added to each analysis to serve as an internal control to verify the accuracy of the analysis.

[0130] Figure 2 shows the analysis of small RNAs by the 2100 Bioanalyzer. A: Lavender extract according to Example 1. B: Conventional lavender extract according to Example 2.

[0131] Biochemical analysis demonstrated that low molecular weight RNA can be extracted from lavender using the method of the present invention, as shown in Figure 2A. Figure 2A shows that the RNA present in the lavender extract of the present invention has a molecular weight ranging from greater than 25 to approximately 150 nucleotides.

[0132] Conventional extraction methods, such as maceration, described in Example 2 below, do not extract low molecular weight RNA (Figure 2B). This analysis also demonstrated that these molecules are not present in either the floral water or the essential oil. This analysis also demonstrated the absence of DNA in the extract.

[0133] Volatile odor compounds (VOCs) were analyzed for the lavender extract, floral water, and essential oil of the present invention by GC-FID. All samples were separated by Agilent GC / FID 7890A gas chromatography (Agilent Technologies) on a 30 m x 250 μm x 0.25 μm GC OPTIMA 5HT column (Macherey-Nagel 726106.30). 3 μL of sample product was injected onto the column at a flow rate of 1.3 ml / min using helium as the carrier gas in a programmable oven ramp from 75°C to 320°C in 40 minutes.

[0134] Hydrogen (30 ml / min) and air (400 ml / min) were used as the flame, and the molecules were detected with a flame ionization detector at 230° C. VOCs were identified by comparing the retention times of the compounds.

[0135] For aqueous samples, a liquid / liquid extraction in hexane (1:1) was performed before injection. Magnesium sulfate was added to the organic phase to remove all water.

[0136] [Table 4]

[0137] The results of the GC-FID analysis shown in Table 4 indicate that the lavender extract of Example 1 does not contain any terpene-based odor molecules, unlike lavender floral water and essential oil, which are known to contain a large amount of these molecules.

[0138] [Example 2] <Preparation of lavender macerate>

[0139] To compare the so-called classic extract with the extract of the present invention, a lavender infusion was prepared using the same amount of dried lavender flowers of the species Lavandula angustifolia as in Example 1, i.e., 3% crushed dried lavender flowers in distilled water. The mixture was then heated at 80°C for 1 hour. After that, to remove solid residual plant matter from the liquid portion, the mixture was filtered first through a large porosity filter of 30 μm, followed by successive filters with decreasing porosity filters down to 0.2 μm. The purpose of this method was to prepare a control extract for obtaining comparative analytical data with the lavender extract obtained by the method of the present invention. The results are shown in Figure 2B and are exemplified herein.

[0140] [Example 3] <Evaluation of the Lavandula angustifolia extract of Example 1 on reactive oxygen species after visible light stress in normal human keratinocytes>

[0141] (principle) The purpose of this study was to demonstrate the effect of lavender extract prepared according to Example 1 on reducing reactive oxygen species generated by visible light stress. This type of light, between 400 and 700 nm, was intended to mimic sunlight. Reactive oxygen species are involved in various mechanisms of protein and DNA changes associated with skin aging.

[0142] (Protocol) Normal human keratinocytes were treated with the extract of Example 1 overnight. Then, the cells were exposed to visible light generated by a 24-watt, 5000 Kelvin spotlight, simulating sunlight. This exposure was repeated four times during the day for 10 minutes. This treatment was repeated again at night, and then the cells were subjected to the same visible light stress again. At the end of this stress, reactive oxygen species were detected using the mitochondrial probe MitoSOX™ Red (ThermoFisher Scientific).

[0143] (result) As a result of applying sunlight stress, the reactive oxygen species (ROS) increased by +43% compared to the non-exposed cells. When the cells were treated with the extract of Example 1 at 0.1% (volume / volume dilution ratio), the ROS decreased very significantly by -12% compared to the untreated cells. As a result of testing the lavender immersion obtained in Example 2 under the same conditions, the decrease was smaller at -5%.

[0144] (Conclusion) The lavender extract showed antioxidant activity directed against mitochondrial-level reactive oxygen species. This activity was found to be higher than that obtained with the conventionally immersed lavender extract.

[0145] [Example 4] <Evaluation of the extract of Example 1 against DNA damage in normal human melanocytes exposed to UVB stress>

[0146] (Principle) Genomic instability can be defined as any chemical modification of DNA that occurs in various processes and can accumulate over time. Changes in DNA are a major factor in photoaging. In this study, we focused on UVB damage in melanocytes. Pyrimidine dimers are formed when UVB acts directly on the pyrimidine bases of DNA. Cyclobutane pyrimidine dimers (CPDs) are formed, which are the most common form of DNA damage induced by UVB. In melanocytes, CPDs continue to be generated for more than 3 hours after UVB exposure. This is referred to as "dark CPD" and is due to the chemical excitation of melanin and the transfer of energy to DNA. In this study, the ability of the extract of Example 1 to reduce the formation of dark CPDs in melanocytes was evaluated.

[0147] (Protocol) Melanocytes extracted from human epidermis were treated overnight with the extract of Example 1 at 0.1% (volume / volume dilution ratio), irradiated with UVB at 60 mJ / cm2, and then treated overnight again with the lavender extract. The next morning, pyrimidine dimers were detected by immunostaining using an antibody against CPD (Cyclobutane Pyrimidine Dimers Mouse Monoclonal, Euromedex). After 1.5 hours of incubation and washing, the cells were incubated in the presence of an anti-mouse secondary antibody conjugated with a fluorescent dye (Alexa Fluor® 488, Invitrogen). Subsequently, the cells were examined with an epifluorescence microscope (Zeiss Axiovert 200M microscope). Then, the presence of CPD was observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0148] (Results) Under non-irradiation conditions, melanocytes did not show CPD. These formations are induced as a result of UVB stress. When the extract of Example 1 was applied to melanocytes, the induction of dark CPD by UVB was reduced by up to -37% (very significant by Student's t-test compared to untreated irradiated cells). At the same conditions and the same concentration of 0.1%, the lavender immersion obtained in Example 2 had no significant effect.

[0149] (Conclusion) The extract of Example 1 reduced the "dark CPD" generated in melanocytes by UVB stress. By this action, it was found that the lavender extract showed the benefit of reducing DNA damage and contributed to the skin repair mechanism at night.

[0150] [Example 5] <Evaluation of the extract of Example 1 on the ex vivo skin biopsy and the melanin synthesis pathway in human hair follicles>

[0151] (Principle) The purpose of this experiment was to demonstrate the effect of the extract of Example 1 on melatonin synthesis in cultured human skin biopsies. The evaluation included two markers: 1) the AANAT enzyme (aralkylamine N-acetyltransferase, or serotonin N-acetyltransferase, or timezyme), which catalyzes the N-acetylation of serotonin to N-acetylserotonin, the final step in melatonin synthesis, and 2) melatonin itself. The AANAT enzyme controls the day / night rhythm of melatonin production in the pineal gland. Because melatonin is also synthesized topically in the skin, it was desirable to monitor melatonin synthesis in response to the application of lavender extract.

[0152] (Protocol) AANAT enzyme and melatonin were evaluated by indirect immunofluorescence in skin biopsies pretreated with topical lavender extract for 48 hours (twice daily). Human hair follicles isolated from scalp biopsies were also exposed to the extract of Example 1 diluted to 0.5% (volume / volume dilution) in culture medium. Control biopsies and control hair follicles incubated in parallel under identical conditions without lavender extract were administered placebo (phosphate-buffered saline, PBS). After incubation, the biopsies and hair follicles were fixed and embedded in paraffin for tissue sectioning. Detection of AANAT enzyme and melatonin was performed by incubation with anti-AANAT (Invitrogen) and anti-melatonin (Abnova, Cliniscience) antibodies, respectively. After 1.5 hours of incubation and washing, the sections were incubated in the presence of a fluorescent dye-conjugated anti-rabbit secondary antibody (Alexa Fluor® 488, Invitrogen). Sections were then examined under an epifluorescence microscope (Zeiss Axiovert 200M microscope). Collagen I expression was then observed and quantified by image analysis (Volocity® image analysis software, Improvision).

[0153] (result) Evaluation of AANAT enzyme and melatonin showed increases of +20% and +51% respectively in skin biopsies treated with the 0.5% extract of Example 1 (which were highly significant by Student's t-test compared to placebo-treated biopsies). The lavender infusion obtained in Example 2 gave equivalent results for AANAT, but the increase in melatonin was smaller (+34%, highly significant by Student's t-test compared to placebo-treated biopsies). In cultured hair follicles, the 0.5% extract of Example 1 brought about a +23% increase in melatonin production observed in the outer epithelial sheath of the hair follicle.

[0154] (Conclusion) The extract of Example 1 showed activity on melatonin production in ex vivo skin biopsies and hair follicles. This increase is related to an increase in the AANAT enzyme whose expression increases in the pineal gland during the transition from day to night. The properties of melatonin are related to its action in repairing cell damage due to its antioxidant activity, particularly in DNA. Therefore, the increase in skin melatonin by the lavender extract seems to be beneficial for the night-time skin damage repair process. The increase in melatonin in hair follicles indicates that it has a beneficial effect on the physiological function of hair follicles since melatonin is related to the growth phase of hair.

[0155] [Example 6] <Evaluation of the Whitening Ability of the Extract of Example 1 in Ex Vivo Skin Biopsies>

[0156] (Principle) The aim of this test was to evaluate the whitening ability of the extract of Example 1 in ex vivo skin biopsies using histological staining of Fontana-Masson for melanin based on the reduction of ammoniacal silver nitrate solution to metallic silver. The obtained staining revealed the melanin content, which was quantified by image analysis.

[0157] (Protocol) Ex vivo human skin biopsies were cultured and treated with 0.5% (volume / volume dilution) and 1% (volume / volume dilution) of the extract of Example 1 for 48 hours. After treatment, the biopsies were fixed and embedded in paraffin for histological analysis. After deparaffinization, sections were incubated in ammoniacal silver nitrate solution at 60°C for 10 minutes. After washing, sections were treated with 5% sodium thiosulfate for 2 minutes, washed again, and mounted for examination under an Eclipse E600 microscope (Nikon). Photographs were taken with a QImaging Retiga 2000R Fast1394 camera and analyzed with Q-Capture Pro7 software (QImaging).

[0158] (Results and Conclusions) In ex vivo skin biopsies, a decrease in melanin content of -55% and -72% was observed after application of the extract of Example 1 at 0.5% (volume / volume dilution) and 1% (volume / volume dilution), respectively (highly significant by Student's t-test compared to placebo biopsies), whereas the lavender infusion obtained in Example 2 showed no decrease at 0.5% and a smaller decrease (-47%) at 1%.

[0159] From this study, it can be concluded that the Lavandula angustifolia extract of Example 1 has potential skin whitening effects on ex vivo skin biopsies.

[0160] [Example 7] <Rich cream formula>

[0161] [Table 5] JPEG0007758679000006.jpg48166

[0162] (Adjustment method) 1. Homogenize Phase A in the main vessel and begin heating to 75-80°C. 2. At 30°C, sprinkle onto phase B and homogenize while heating. 3. Prepare Phase C in a separate beaker and heat to 75-80°C until homogeneous. 4. At 75°C, Phase C was added to the main vessel and homogenized for 10 minutes. 5. Allow temperature to cool and add Phase D at 65°C. Mix thoroughly and homogenize for 10 minutes. 6. Phase E was premixed and then added to the main vessel. 7. At 60°C, add Phase E. Mix thoroughly and homogenize for 10 minutes. 8. At 35°C, Phase F was premixed and added and mixed thoroughly. 9. Phase G was premixed and then added to the main vessel. 10. At 35°C, add Phase G. Mix thoroughly to homogenize. 11. Phase H was prepared in a separate beaker by sprinkling Natrosol™ into room temperature water and homogenizing while heating to 60°C. 12. At 30°C, add Phase H. Mix thoroughly to homogenize. 13. It stopped at 25°C.

[0163] The composition was in the form of a pale pink buttercream, had a pH between 4.90 and 5.40, and a viscosity (D0) of 160,000-210,000 cps (Brookfield RVT / spindle D / 5 RPM / 1 min / 25°C).

[0164] [Example 8] <Anti-aging mask formula>

[0165] [Table 6]

[0166] (Adjustment method) 1. Homogenize Phase A in the main vessel at 25°C. 2. At 25°C, sprinkle onto Phase B and mix thoroughly until uniform. 3. At 25°C, Phase C was added and mixed thoroughly until uniform. 4. Phase D was premixed in a separate beaker and added to the main vessel at 25°C. 5. At 25°C, Phase E was added to the main vessel and mixed thoroughly. 6. Phase F was premixed and slowly added. Mix thoroughly until uniform. 7. Phase G was premixed in a separate beaker and added to the main vessel until uniform. 8. It stopped at 25°C.

[0167] The composition appeared as a creamy gel with a brilliant green effect, a pH between 5.30 and 5.80, and a viscosity (D0) of 70,000-100,000 cps (Brookfield RVT / spindle C / 5 RPM / 1 min / 25°C).

[0168] [Example 9] <Beauty serum formulation>

[0169] [Table 7]

[0170] (Adjustment method) 1. Water was added to the main vessel and mixing was initiated with the high-low propeller blades. 2. The remaining ingredients were added in order, mixing after each addition.

[0171] This composition was a smooth, translucent serum with a pH between 5.75 and 6.25 and a viscosity (D0) of 1,100 to 1,400 cps (Brookfield RVT / spindle 3 / 20 rpm / 25°C / 1 minute).

Claims

1. 1. A method for obtaining an aqueous extract of the aerial parts of lavender, comprising the steps of: a) contacting the aerial parts of lavender with water; b) adding phytic acid to the mixture obtained in a) at a concentration between 1 and 10 mM and at a pH between 10 and 11; c) then adjusting the pH of the mixture obtained in b) to a value between 6 and 8; d) purifying the mixture obtained in c) to remove residual solid plant matter to obtain a purified aqueous crude extract; e) checking the pH and readjusting it to a value between 6 and 8; A method comprising:

2. 2. The method of claim 1, wherein in step e) the pH is readjusted to a value between 6 and 6.

5.

3. 3. The method according to claim 1 or 2, wherein in step a) the previously dried and then crushed aerial parts of lavender are contacted with water in a plant material / water ratio of between 3 and 20% (w / w).

4. 4. The method according to any one of claims 1 to 3, wherein the treatment with phytic acid at a concentration of 3 mM in step b) is carried out for at least 1 hour and at a temperature between 20 and 80°C with stirring.

5. 5. The process according to claim 1, wherein step e) is preceded by at least one filtration of the aqueous crude extract obtained in d).

6. 5. The method according to any one of claims 1 to 4, wherein prior to step e), the crude aqueous extract obtained in d) is subjected to successive filtrations by decreasing the filtration threshold from 20-50 μm to 0.10-0.30 μm.

7. 7. The method according to any one of claims 1 to 6, wherein the lavender aerial parts are the aerial parts of the species Lavandula angustifolia.

8. 8. An aqueous extract of the aerial parts of lavender obtained by the method according to any one of claims 1 to 7, which is free of DNA and enriched in small RNAs up to 150 nucleotides in length, sugars, phenolic compounds and organic acids, the aqueous extract containing 2-10 g / kg of sugars, 100-1500 mg / kg of organic acids, 500-2000 mg / kg of phenolic compounds and 40-200 mg / kg of small RNAs up to 150 nucleotides in length, based on the total weight of the extract, and comprising 10-30 g / kg of dry weight.

9. A composition comprising the extract of claim 8 and a solvent, the composition comprising, by weight relative to the total weight of the composition, 4 to 20 g / kg of dry extract, 0.5 to 10 g / kg of sugars, 50 to 700 mg / kg of organic acids, 50 to 1500 mg / kg of phenolic compounds, and 10 to 100 mg / kg of small RNAs up to 150 nucleotides in length.

10. A composition comprising, as an active ingredient, an effective amount of the composition of claim 9 and a physiologically acceptable medium.

11. 11. The composition according to claim 10, wherein the extract is present in a concentration of between 0.05 and 5% by weight relative to the total weight of the composition.

12. 12. The composition of claim 11, wherein the extract is present in a concentration of between 0.1 and 1.0% by weight relative to the total weight of the composition.

13. A composition according to any one of claims 10 to 12, formulated for topical application to the skin, skin appendages and scalp.

14. 14. The composition according to any one of claims 10 to 13 for skin care, for care of the scalp and skin appendages, for protecting the skin from external aggressions and oxidation, for preventing the signs of skin ageing, for improving photoprotection, for whitening the skin, for improving skin moisture retention, for strengthening the barrier function or for soothing the skin.

15. A composition according to any one of claims 10 to 13 for improving the biological mechanisms involved in skin repair during the night.

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