Use of a plaitleaf milkweed extract to promote hair growth

Abstract

Use of a plant extract of a species of the genus Fimbristylis for promoting and / or increasing hair growth, and / or preventing hair loss, and / or increasing the thickness of the hair, in particular the eyelashes and / or eyebrows. Also disclosed are methods of preparing the plant extract and compositions comprising the plant extract.

Description

Technical Field

[0001] This invention relates to the use of plant extracts from species of the genus *Myrothamnus* SP. for promoting hair growth and / or preventing hair loss, and / or increasing hair thickness. In one embodiment, the hair is eyebrow and / or eyelash hair. Methods for preparing the plant extracts and compositions comprising the plant extracts are also disclosed. Background Technology

[0002] Hair, especially eyelashes and eyebrows, plays a vital role in human aesthetics. Eyelashes and eyebrows are prominent facial features. They protect the eyes and influence facial expressions. These hairs grow relatively slowly and are on average short. In many cultures, long, thick eyelashes are particularly valued and admired. People are interested in enhancing the appearance of eyelashes by increasing their length and thickness. Similarly, there is interest in enhancing the appearance of eyebrows by increasing their thickness.

[0003] Hair damage and / or loss is a common problem in human subjects and can significantly impact a person's overall appearance. Eyelashes and eyebrows are particularly susceptible to damage from finger / hand contact, such as when subjects rub their eyes when fatigued. Mechanical or chemical stresses during styling can weaken and damage hair. Hair loss can have numerous other causes (e.g., genetic predisposition, aging, and / or disease). There is considerable interest in hair care methods that can increase hair strength, prevent hair loss, or promote hair growth.

[0004] Hair grows from hair follicles in the skin, which extend from the epidermis (the outermost layer of skin) to the dermis (the deepest layer of skin). The length and thickness of hair depend primarily on two biological processes: hair follicle regeneration and hair fiber synthesis.

[0005] The hair cycle is a stem cell-mediated process that occurs in adult skin, involving the periodic destruction and regeneration of hair follicles. The hair cycle consists of three defined phases: the anagen (growth phase), followed by the catagen (transitional) phase and the telogen (resting) phase. The anagen phase is the longest phase of the hair cycle. During this phase, cells begin to proliferate, producing hair follicles, and hair continuously forms. In the catagen phase, proliferation ceases, and apoptosis increases. Hair growth stops, and the hair follicle shortens. Finally, in the telogen phase, described as the resting phase, hair breaks down. [Alonso L and Fuchs E (2006) The hair cycle. J. Cell Sci. 119:391-393.]

[0006] New hair growth requires re-entry into the early stages of hair growth, a process involving the activation of pluripotent epithelial stem cells in a specialized region of the outer root sheath (ORS) of the hair follicle called the ridge. Dermal cells called follicular dermal papilla cells (HFDPCs) are located in the ridge [Beaudoin GM 3rd, Sisk JM, Coulombe PA and Thompson CC. (2005) Hairless triggers reactivation of hair growth by promoting Wnt signaling. ProcNatl Acad Sci USA. 102:14653–14658]. HFDPCs in adult hair follicles play a crucial role in the hair growth cycle by inducing hair follicle development [Driskell RR, Clavel C, Rendl M and Watt FM. (2011) Hairfollicle dermal papilla cells at a glance. J.Cell Sci. 124:1179-1182].

[0007] Hair fibers are primarily composed of protein (accounting for 65% to 95% of hair weight), with keratin being the most abundant. This fiber consists of three common layers: the cuticle, the cortex, and the medulla. The central medulla contains polygonal cells with a spongy appearance. Surrounding the medulla is a layer of keratinized fibroblasts in the cortex, which are longitudinally oriented and wrapped in keratin filaments. Additionally, the cortex contains melanosomes that determine the color of the hair fiber. The outermost layer of the hair fiber is the cuticle, which consists of multiple layers of keratinocytes. The cuticle is thin and translucent, allowing light to penetrate the pigment in the cortex. The keratin that forms this complex structure of the hair fiber is a result of the proliferative activity of keratinocytes located in the hair follicle. These keratinocytes, present in the dermal papilla (DP), along with other cells involved in this proliferative activity, promote hair growth.

[0008] Compositions and methods for enhancing hair growth are known in the art. However, known methods may be ineffective or may have undesirable side effects.

[0009] Drugs, including (Luo Jian) (Propecia) and (Avodat) are all approved treatments for hair loss. However, the efficacy of these drugs is primarily limited to hormone-related alopecia (i.e., androgenetic alopecia) and requires a medical prescription. The use of synthetic chemicals in cosmetic formulations intended for human use is also raising growing concerns. Developing cosmetic formulations for areas close to the eyes (such as those designed to promote the growth of eyelashes or eyebrows) presents particular challenges because the human eyeball and surrounding membranes are especially sensitive compared to human skin in other areas of the body. Although eyelash cosmetics are not intended for direct application to the eyeball or cornea, users may accidentally transfer some cosmetic formulation into the eyeball during application. Furthermore, there is growing concern about the environmental impact of the production of synthetic chemicals used in cosmetic products, for example, due to washing cosmetics off the body after use or due to the disposal of many unused such products in landfills.

[0010] Volumizing mascara is one of the most common products for enhancing the appearance of eyelashes. However, those with short eyelashes may find it difficult to achieve striking lashes with mascara alone, and may opt for eyelash extensions. Eyelash extensions typically involve using inferior adhesives to attach the extensions to the area near the upper lash line on the eyelid, potentially damaging the original eyelashes. Various risks associated with the use of eyelash extensions have been reported, such as eyelid swelling, eyelash loss, eyelid skin irritation, or eyelid infection. There is a need for a mascara composition that can enhance the appearance of eyelashes and promote eyelash growth.

[0011] Micropigmentation and microblading are eyebrow tattooing procedures used to enhance the density and definition of eyebrows. However, they bring the inconveniences associated with any tattooing procedure, such as pain and the risk of infection.

[0012] There is a need for natural, sustainable, and effective solutions to address the problems identified above. Several herbal preparations have been proposed to promote eyelash and eyebrow hair growth; however, the effectiveness of these preparations may be limited.

[0013] EP2764894B1 provides a blackberry leaf extract for inducing hair growth. This extract is claimed to initially induce a pronounced telogen effluvium (i.e., clubbed hair loss), followed by a rapid transition to telogen effluvium (i.e., the active growth phase of the hair follicle). Extractants used to obtain the extract include ethanol, or combinations of ethanol and alcohols. Extraction with ethanol and alcohols presents inherent flammability hazards in manufacturing, production, and transportation logistics. While ethanol is considered a green solvent with good extraction capabilities, certain segments of the cosmetics market and some consumers prefer not to use it.

[0014] US20190224160A1 discloses a composition for promoting eyelash growth, comprising at least one keratin derived entirely from Coleus forskohlii and a keratinocyte growth stimulant used in combination therewith. The keratinocyte growth stimulant comprises at least one plant extract selected from the group consisting of: Tussilago farfara flower extract, Achillea millefolium extract, Cinchonas uccirubra bark extract, Nasturtium officinale, and Tropaeolum majus.

[0015] WO2017032711 discloses a cosmetic composition for promoting eyelash growth, the composition comprising a plant extract of mung bean (Vigna radiata), an extract of mung bean sprouts, and a peptide having an amino acid sequence of Gly-His-Lys from the N-terminus to the C-terminus.

[0016] *Flavouria* species are drought-tolerant shrubs native to South Africa, belonging to the plant family known as "resurrection plants." These plants have adapted to the region's seasonal wet and dry seasons. During the dry season, the plant enters a dormant state while awaiting rain, its leaves drying out. Upon the first rainfall, the plant revives and flourishes, its leaves rapidly absorbing water, followed by flowering. Resurrection plants possess multiple pathways to cope with mechanical and drought stress, attracting the interest of the scientific community seeking active substances with cosmetic and / or pharmaceutical properties. *Flavouria* is consumed as a tea in South Africa and its essential oils are also extracted. Its use in local traditional foods and medicines is well-known in South Africa. *Flavouria* is already used in the cosmetics industry for skincare and is the subject of several patents / patent applications, such as US200701341934A1, FR29978536B1, and KR1305698B1.

[0017] There is a need to find natural, non-therapeutic cosmetic care methods that can increase hair growth and thickness, especially those related to hair in sensitive facial areas (such as around the eyes), i.e., eyelashes and / or eyebrows. Ideally, such care methods will have few or no negative side effects and / or provide aesthetic benefits to the hair and the associated skin.

[0018] This invention addresses some or all of the requirements identified above, and solves some or all of the problems identified above. Summary of the Invention

[0019] In a first aspect, the present invention provides the following uses of extracts from *Flavorum* species: promoting hair growth; preventing hair loss; and / or increasing hair thickness. In particular, this use can be for non-therapeutic cosmetic purposes. Specifically, the hair can be eyelash and / or eyebrow hair. The present invention provides non-therapeutic cosmetic uses of *Flavorum* species extracts for increasing eyelash length and / or increasing eyebrow thickness (i.e., increasing hair density). The use of *Flavorum* extracts is particularly advantageous because consumers are particularly interested in beauty treatments considered "natural," as such treatments are generally more environmentally friendly.

[0020] On another front, the present invention provides a method for promoting and / or increasing hair growth; preventing hair loss; and / or increasing hair thickness, comprising applying a plant extract of a species of the genus *Pterocarya* to a subject. Specifically, this method can be a non-therapeutic cosmetic procedure. Specifically, the hair can be eyelash or eyebrow hair. The present invention provides a non-therapeutic cosmetic method for increasing eyelash length and / or increasing eyebrow thickness (i.e., increasing hair density), comprising applying a plant extract of a species of the genus *Pterocarya* to a subject.

[0021] In another aspect, the present invention provides a method for obtaining plant extracts from species of the genus *Pycnogenolus*, comprising extracting plant material of the genus *Pycnogenolus* using subcritical water, wherein the extraction includes the following steps:

[0022] i) To form an aqueous plant extract, the plant material is brought into contact with subcritical water for at least 10 minutes at a temperature of at least about 120°C and a pressure suitable for maintaining the water in a liquid state.

[0023] as well as

[0024] ii) Isolation of plant materials from aqueous plant extracts.

[0025] In another aspect, the present invention provides an aqueous plant extract that can be obtained by / through the aforementioned method of obtaining plant extracts from species of the genus *Pyrrosia*.

[0026] In another aspect, the present invention provides a cosmetic composition comprising an aqueous plant extract and at least one cosmetically acceptable excipient or ingredient.

[0027] In another aspect, the present invention provides a mascara composition comprising a plant extract from a species of the genus *Pterocarya* and at least one pigment. Detailed Implementation

[0028] This invention is based on the discovery that plant extracts from the genus *Flavorum* (also known as *Flavorum* species) possess some surprising properties that make the extracts usable for non-therapeutic cosmetic applications.

[0029] definition

[0030] In the context of this invention, "hair" includes hair on the subject's scalp, skin, eyelashes, eyebrows, mustache area, and / or beard area. Specifically, hair refers to the hair in the eyebrows and / or eyelashes. "Skin" should be understood as the multiple layers constituting the skin, from the outermost layer, the stratum corneum, to the innermost layer, the subcutaneous tissue (both included). These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, mast cells, neurons, and / or fat cells. The term "skin" includes the skin of mammals, such as human skin, and includes skin with hair.

[0031] As used herein, the term “about”, for example when referring to a measurable value (such as the amount or weight of a particular component or temperature), means a variation of ±20%, ±10%, ±5%, ±1%, ±0.5%, or particularly ±0.1% of the specified amount.

[0032] As used herein, the terms “comprising” or “including” (which are inclusive or open-ended and do not exclude additional unlisted elements or method steps) are intended to cover, in alternative embodiments, the phrases “consistently made of” and “composed of”, wherein “consistently made of” excludes any unspecified elements or steps and “consistently made of” allows the inclusion of additional unlisted elements or steps that do not substantially affect the nature or essential and novel features of the composition or method under consideration.

[0033] extract

[0034] This invention relates to the use of plant extracts from species of the genus *Flavourae*. The terms "plant extract" and "extract" are used interchangeably herein and refer to a product containing one or more compounds that have been extracted from plant material of a species of the genus *Flavourae*. Specifically, the product is obtained by solid / liquid extraction of plant material of a species of the genus *Flavourae*, thereby extracting one or more phytochemical compounds contained in the plant material (solid) from the plant material into a solvent (liquid); these phytochemical compounds dissolve from the plant material and enter the solvent. The product may be a composition comprising an extraction solvent and one or more phytochemical compounds. The solid / liquid extraction method may involve a separation step, optionally followed by one or more purification steps. Thus, the extract may be a product obtained after a separation step or a product obtained after a purification step. In the separation step, a composition comprising a solvent and one or more phytochemical compounds is separated from the plant material. The extract is the separated composition comprising an extraction solvent and one or more phytochemical compounds. The solubility of the phytochemical compounds in the solvent may be temperature-dependent, therefore the composition may be a solution, or some phytochemical compounds may precipitate if it is cooled below the temperature used for extraction. When the solvent is water, a composition comprising an extraction solvent and one or more phytochemical compounds is also referred to herein as an "aqueous plant extract" or "aqueous extract". Purification of a composition comprising a solvent and one or more phytochemical compounds (e.g., by fractionation, concentration, or drying) provides an extract that may be, for example, a concentrate (e.g., in a viscous form) or a solid (e.g., a powder).

[0035] The genus *M. flabellifolia* belongs to the genus of small, drought-tolerant flowering shrubs and includes two species: *M. flabellifolia* and *M. moschata*. Specifically, extracts can be obtained from *M. flabellifolia*. *M. flabellifolia* is also referred to as *M. flabellifolius* in the literature.

[0036] Typically, plant material from species of the genus *Flavorata* comprises the above-ground parts of the plant, particularly the stems and / or leaves. Plant material may contain stems and / or leaves as the main component, for example, stems and / or leaves accounting for more than 50%, 80%, or 90% of the weight of the plant material. Plant material may also contain only stems and / or leaves, i.e., stems and / or leaves are the sole component, with no other plant material present.

[0037] Plant materials are typically sold in dried form. Drying plant materials makes them easier to homogenize, handle, and store. "Dried plant material" refers to plant material with a moisture content of less than 10% by weight, less than 5% by weight, less than 2% by weight, or less than 1% by weight. For example, moisture content can be measured using the official AOAC (Association of Official Analytical Chemists) method AOAC 934.06-1934 (1996). However, undried plant material can also be used.

[0038] Advantageously, the plant material is ground or crushed prior to extraction. The ground plant material can have a particle size, for example, ranging from 100 μm to 50 mm, with an average particle size of 0.01 mm or 0.1 mm to 10 mm. More particularly, the average particle size can range from 0.2 mm to 5 mm. The average particle size can be determined by conventional methods, such as those involving sieving analysis. Any suitable grinding / crushing technique known in the art can be used to obtain plant material with the desired particle size. Particularly suitable plant material is dried plant material in powder form having, for example, the particle sizes mentioned above.

[0039] Plant materials can be mixed with neutral materials to make them drier and / or more porous, facilitating solvent extraction. Suitable neutral materials include graphene, silica gel, C18 resin, diatomaceous earth, and neutral alumina.

[0040] Suitable solvents for extraction include water, lower alcohols containing one to four carbon atoms (e.g., methanol, ethanol, butanol, etc.) and diols, as well as combinations thereof. Advantageously, extracts from *Pyrantelia* species can be obtained using water as the sole extraction solvent. In this case, compounds in the plant material dissolve from the plant material and enter the water to form an aqueous extract (also referred to herein as an aqueous plant extract). For example, the aqueous extract can be purified to form an extract as a concentrate or powder. Therefore, it is advantageous to obtain extracts from *Pyrantelia* species by avoiding the use of organic solvents during the extraction process.

[0041] Any conventional solid / liquid extraction method can be used to obtain plant extracts from plant material of *Pycnogenolus* species, such as Soxhlet extraction, percolation, and maceration. The temperature of the solvent should be selected to suit the solvent. If water is used as the extraction solvent, it is preferably used at a temperature above 60°C. Typically, the ratio of dried plant material to solvent (weight of dried plant material (g) / volume of solvent (mL)) is in the range of 1:5 to 1:50, or 1:10 to 1:30, or 1:15 to 1:25, or 1:5 to 1:15. In particular, this ratio is 1:10 or 1:20.

[0042] In one embodiment, extracts from *Pycnogenolus* species are obtained by solid / liquid extraction of plant material from *Pycnogenolus* species using subcritical water as the extraction solvent. Subcritical water is water that is kept liquid under pressure at a temperature above its natural boiling point (i.e., above its boiling point at atmospheric pressure) of 100°C. Subcritical water can have a critical point temperature as high as 374°C. Subcritical water is also referred to as “pressurized low-polarity water,” “pressurized hot water,” or “compressed hot water.” Heating water under pressure to a temperature above its boiling point causes a change in its key properties, such as polarity. Preferably, when subcritical water is used as the extraction solvent, it is the only extraction solvent used in the method; that is, the extraction is carried out with subcritical water in the absence of any other solvent (organic or inorganic).

[0043] More specifically, extracts from species of the genus *Flavourae* can be obtained by extracting plant material of *Flavourae* species with subcritical water, wherein the extraction includes contacting the plant material with subcritical water at a temperature of at least about 120°C and a period of time suitable for maintaining the water at a liquid pressure for at least 10 minutes to form an aqueous plant extract. More specifically, extracts from species of the genus *Flavourae* can be obtained by a method including the following steps:

[0044] i) To form an aqueous plant extract by contacting plant material of the genus *Pterocarya* with subcritical water for at least 10 minutes at a temperature of at least 120°C and a temperature and pressure suitable for maintaining the water in a liquid state; and

[0045] ii) Isolation of plant materials from aqueous plant extracts.

[0046] Typically, the ratio of dried plant material to subcritical water (weight of dried plant material (g) / volume of water (mL)) is in the range of 1:5 to 1:50, or 1:10 to 1:30, or 1:15 to 1:25, or 1:5 to 1:15. In particular, this ratio can be 1:10 or 1:20.

[0047] Specifically, extraction can be carried out using subcritical water at temperatures of about 120°C to about 220°C, about 130°C to about 180°C, about 140°C to about 160°C, or about 145°C to about 155°C. Specifically, subcritical water can be at a temperature of about 150°C.

[0048] When subcritical water is used as the extraction solvent, extraction is carried out in a pressurized vessel (typically a stainless steel container). During extraction, the pressure in the pressurized vessel must be sufficient to maintain the subcritical water in a liquid state. The pressure required to achieve this will vary depending on the water temperature. Those skilled in the art will be able to determine the required pressure. Typically, the pressure ranges from 0.5 MPa to 20 MPa. The pressure can be at least about 1 MPa. The pressure can be about 5 MPa to about 15 MPa, or about 8 MPa to about 13 MPa. The pressure can be 10 MPa, 11 MPa, and 12 MPa.

[0049] The step of contacting the plant material with subcritical water shall be carried out for a period of at least about 10 minutes. This period is the time during which the plant material is in contact with the subcritical water, whether the water is static or flowing over the plant material. This period is also referred to herein as the extraction time. The extraction time will vary, for example, depending on the amount of plant material and water used. Typically, the extraction time will range from 10 minutes to about 5 hours. For example, the extraction time may be at least about 15 minutes, or at least about 30 minutes, or at least about 1 hour, or at least about 2 hours, or at least about 3 hours. In particular, the extraction time is from about 10 minutes to about 2 hours, and more particularly from 20 minutes to 90 minutes. The extraction time may be from 45 minutes to 75 minutes, or about 1 hour.

[0050] Subcritical water extraction can be performed in batch mode (also known as "static mode") or dynamic mode (also known as "flow-through"). In batch mode, plant material is exposed (i.e., contacted) with subcritical water in batches. In batch mode, the volume of subcritical water used is the total volume of subcritical water used for all batches of the plant material sample. In batch mode, the extraction time is the total time the plant material is exposed to subcritical water in all batches of the plant material sample. In dynamic mode, the plant material is exposed to a continuous flow of water. In dynamic mode, the volume of subcritical water used is the total volume of subcritical water used for the plant material sample. In dynamic mode, the extraction time is the total time the plant material is exposed to the subcritical water flow used for the plant material sample. For faster and larger-scale production of plant extracts, extraction is advantageously performed in dynamic mode. Extraction can be performed in any system known in the art that allows subcritical water extraction, including systems with batch or continuous extractors.

[0051] In the step of contacting plant material with subcritical water, compounds are extracted from the plant material into the subcritical water. More specifically, components of the plant material are dissolved in subcritical water to form an aqueous plant extract. Similarly, conventional solvent extraction methods using water as a solvent will form an aqueous plant extract. This solvent extraction method may include the step of separating the plant material (i.e., any undissolved plant material) from the plant extract. Typically, this step is straightforward. For example, in the case of subcritical water extraction, this step may simply involve releasing the aqueous plant extract from a pressurized container while retaining the plant material in the pressurized container. Other suitable separation techniques are known in the art and include, for example, filtration, sedimentation, decantation, or centrifugation. Filtration can be performed using filters with pore sizes less than 1000 μm, or less than 20 μm, or less than 10 μm, or less than 1 μm, or less than 0.1 μm. Filtration can be performed in a continuous filtration operation, for example using filters with gradually decreasing pore sizes. The residue remaining after filtration (undissolved plant material) can be contacted again with more extraction solvent. This filtration-re-contact step can be performed once or repeated, for example, 1 to 5 times. In the case of subcritical water extraction, preferably, separation is performed before the aqueous plant extract is cooled and the extract components are precipitated.

[0052] After plant extracts have been isolated from plant material, they can be purified. In the context of this invention, “purification” means purification, partial purification, and / or fractionation. Numerous techniques well known in the art exist for purifying plant extracts. Some non-limiting examples include solid-liquid extraction, liquid-liquid extraction, solid-phase extraction (SPE), membrane filtration, ultrafiltration, dialysis, electrophoresis, solvent concentration, centrifugation, ultracentrifugation, liquid or gas chromatography with or without high pressure (including size exclusion, affinity, etc.), lyophilization, evaporation, precipitation with various “carriers” (including PVPP, carbon, antibodies, etc.), or various combinations thereof. In one embodiment, the plant extract is concentrated to form a concentrate of the plant extract. Alternatively, the plant extract is dried to form a solid form of the plant extract. The plant extract can be dried to contain no more than 10% by weight of water, no more than 5% by weight of water, no more than 2% by weight of water, or no more than 1% by weight of water. This can be measured, for example, by the moisture content determination AOAC (2000). Suitable concentration and / or drying methods are well known in the art. Examples include, but are not limited to, vacuum evaporation, evaporation, vacuum distillation, distillation, oven drying, sun drying and freeze drying (i.e., lyophilization), spray drying, atomization, or fluidized bed dryers. Plant extracts can be concentrated or dried with or without a carrier or other excipients. The extract can be a concentrate or a solid, such as an amorphous solid, a crystalline solid, or a partially crystalline solid, optionally in powder form.

[0053] Therefore, the present invention also provides plant extracts of *Flavourae* species obtained by / through any of the methods described above. In particular, plant extracts of *Flavourae* species are obtained by / through any of the methods described above involving extraction with water. Plant extracts of *Flavourae* species are obtained by / through any of the methods described above involving extraction with subcritical water. Details of the extraction methods described herein may be applied to or incorporated into the definition of plant extracts provided herein.

[0054] Plant extracts include phytochemicals, which are chemical compounds produced by plants, and may include, for example, polyphenols, amino acids, organic acids, and sugars.

[0055] This plant extract may contain: mirtrin and kaempferol-3-O-glucuronide; or mirtrin, kaempferol-3-O-glucuronide and trehalose; or mirtrin, kaempferol-3-O-glucuronide, trehalose and luteolin-7-O-glucuronide; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide and arbutin; or mirtrin, kaempferol- 3-O-glucuronic acid, trehalose, luteolin-7-O-glucuronic acid, arbutin, and kaempferol-3-O-glucoside; or miguirin, kaempferol-3-O-glucuronic acid, trehalose, luteolin-7-O-glucuronic acid, arbutin, kaempferol-3-O-glucoside, and quercetin-3-O-galactoside; or miguirin, kaempferol-3-O-glucuronic acid, trehalose, luteolin-7-O-glucuronic acid, arbutin, kaempferol-3-O-glucoside, and quercetin-3-O-galactoside; or miguirin, kaempferol-3-O-glucuronic acid, trehalose, and luteolin-7-O-glucoside. -O-glucuronic acid, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, and syringic acid; or miguirin, kaempferol-3-O-glucuronic acid, trehalose, luteolin-7-O-glucuronic acid, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, and quercetin; or miguirin, kaempferol-3-O-glucoside, trehalose, Luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercetin and isoquercetin; or miguirin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercetin, isoquercetin and tryptamine.

[0056] This plant extract can be obtained by solid / liquid extraction using water as a solvent, and may contain: mirtrin and kaempferol-3-O-glucuronide; or mirtrin, kaempferol-3-O-glucuronide and trehalose; or mirtrin, kaempferol-3-O-glucuronide, trehalose and luteolin-7-O-glucuronide; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide. - Glucoside and arbutin; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin and kaempferol-3-O-glucoside; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside and quercetin-3-O-galactoside; or mirtinoside, kaempferol-3-O-glucoside Glucoside, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, and syringic acid; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, and quercetin; or mirtinoside, kaempferol-3-O-glucoside Glucoside, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercetin and isoquercetin; or miguirin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercetin, isoquercetin and tryptophan.

[0057] The plant extract may contain: mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin. -O-glucoside, naringenin and spruce neoglycoside; or miguirin, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside and coniferaldehyde; or miguirin, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside, coniferaldehyde and naringenin-7-O-glucoside.

[0058] This plant extract can be obtained by solid / liquid extraction using water as a solvent, and may contain: mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucoside, and naringenin. -O-glucuronic acid, isorhamnetin-3-O-glucoside, naringenin, and sprucetin; or miguirin, kaempferol-3-O-glucuronic acid, trehalose, luteolin-7-O-glucuronic acid, isorhamnetin-3-O-glucoside, naringenin, sprucetin, and coniferaldehyde; or miguirin, kaempferol-3-O-glucuronic acid, trehalose, luteolin-7-O-glucuronic acid, isorhamnetin-3-O-glucoside, naringenin, sprucetin, coniferaldehyde, and naringenin-7-O-glucoside.

[0059] This plant extract may contain: mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and isorhamnetin-3-O-glucoside; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and isorhamnetin. -3-O-glucoside, naringenin and spruce neoglycoside; or miguirin, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, arbutin, isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside and coniferaldehyde; or miguirin, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, arbutin, isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside, coniferaldehyde and naringenin-7-O-glucoside.

[0060] This plant extract can be obtained by solid / liquid extraction using water as a solvent, and may contain: mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and isorhamnetin-3-O-glucoside; or mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, and naringenin; or mirtinoin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and naringenin. - Glucuronyl glycoside, arbutin, isorhamnetin-3-O-glucoside, naringenin and sprucein; or mirtinoside, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, arbutin, isorhamnetin-3-O-glucoside, naringenin, sprucein and coniferaldehyde; or mirtinoside, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, arbutin, isorhamnetin-3-O-glucoside, naringenin, sprucein, coniferaldehyde and naringenin-7-O-glucoside.

[0061] Composition

[0062] As described above, plant extracts obtained from *Pycnogenolus* species by water extraction can be in an aqueous form, i.e., in the form of aqueous plant extracts. Components extracted from the plant material may or may not be completely soluble in the aqueous plant extract. Advantageously, the aqueous plant extract can be mixed with a water-miscible organic solvent to retain the extracted components in solution. A method for obtaining the plant extracts of the present invention may include the step of mixing an aqueous plant extract as defined herein with a water-miscible organic solvent. The resulting composition comprises the aqueous plant extract and the water-miscible organic solvent. For example, water may be added to the composition to obtain a desired concentration of water-miscible solvent. A method for obtaining plant extracts from *Pycnogenolus* species described herein may include the step of mixing a plant extract as defined herein with a water-miscible organic solvent and optionally water. The resulting composition comprises the plant extract, the water-miscible organic solvent, and optionally water. These solutions can be used as “stock solutions” and can be used, for example, in the preparation of cosmetic compositions.

[0063] The water-miscible organic solvent is preferably a cosmetically acceptable organic solvent and may be a polyol. Suitable polyols include diols, which are organic compounds containing two alcohol functional groups (-OH groups), such as: C2-C10 aliphatic hydrocarbon diols or triols, glycerol. C2-C10 aliphatic hydrocarbon diols include each isomer of C2-C10 or C2-C8 alkyl diols, and they may be substituted or unsubstituted. When substituted, the alkyl diol may be substituted by one or more substituents independently selected from, for example, halogens, hydroxyl groups, esters, nitro groups, cyano groups, haloalkyl groups, sulfonyl groups, and carbonyl groups. The water-miscible organic solvent may be selected from: 1,2-propanediol, ethylene glycol, diethylene glycol, propylene glycol, dipropylene glycol, butanediol, pentanediol, glycerol, or octanediol, and mixtures thereof. In particular, the polyol is glycerol (also referred to herein as glycerol or glycerol glycerol).

[0064] Compositions comprising plant extracts and polyols, and optionally (additional) water, may have a polyol concentration of at least 50% by weight, or 50% to 90% by weight, or 50% to 80% by weight, or 55% to 80% by weight, based on the total weight of the composition. Typically, the balance consists of plant extracts and water. For example, the composition may contain up to 50% by weight of water. Preferably, the polyol is glycerol.

[0065] The extract as described above, or a composition comprising the extract, a water-miscible organic solvent, and optionally water, may be incorporated into a composition suitable for administration to a subject.

[0066] As described above, the composition may be a cosmetic composition comprising an extract or a composition containing the extract, a water-miscible organic solvent, and optionally water, together with at least one cosmetically acceptable excipient or adjuvant. These compositions may be prepared by conventional methods known to those skilled in the art [“Harry’s Cosmeticology”, 7th edition, (1982), edited by Wilkinson JB and Moore RJ, Longman House, Essex, GB].

[0067] The cosmetic composition contains a cosmetically effective amount of plant extract that should be applied. The effective amount of the plant extract and its dosage will depend on a variety of factors, including age, patient condition, the nature or severity of the condition to be treated and / or cared for, the route and frequency of application, and the specific nature of the compound to be used.

[0068] The term "cosmetically effective amount" should be understood to mean an amount of the extract of the present invention that is non-toxic but sufficient to provide the desired effect. The extract or stock solution of the present invention is used in the cosmetic compositions of the present invention at a cosmetically effective concentration to achieve the desired effect; for example, the amount used relative to the total weight of the composition is: 0.00000001% (by weight) to 20% (by weight); 0.000001% (by weight) to 15% (by weight); 0.00001% (by weight) to 10% (by weight); or 0.0001% (by weight) to 5% (by weight); or 0.1% to 4% (by weight); or 1% to 3% (by weight).

[0069] In one embodiment, the extract as described above, or a composition comprising the extract, a water-miscible organic solvent, and optionally water, is present in the cosmetic composition in an amount of 2% (by weight).

[0070] The cosmetic compositions of the present invention can be compositions for topical application, optionally comprising cosmetically or pharmaceutically acceptable excipients necessary for formulation into the desired application form. Topical compositions are those suitable for topical application to mammalian keratinized tissues (such as hair-bearing skin, particularly the human scalp). In particular, topical compositions are hair care compositions, such as conditioning agents, hair growth agents, styling gels, mousses, shampoos, hairsprays, hair oils, styling liquids, coloring, and long-lasting perm compositions. For the purposes of this invention, hair growth agents, conditioning agents, hair growth agents, and styling gels are of particular interest; they can be in the form of gels, lotions, tinctures, sprays, mousses, cleansing compositions, or foams, and can be applied according to individual needs, for example, once daily as a lotion, tincture, mousse, or spray; or once or twice weekly as a conditioning agent or hair growth agent. In particular, suitable compositions in the context of this invention are lotions, shampoos, serums, or mascaras. Preferably, the cosmetic composition is a serum or mascara. The term "serum" is well known to those skilled in the art and refers to a clarified gel-based composition or liquid composition. Serums have a fluid texture and contain a higher concentration of active agents than standard care products. As used herein, the term "mascara" refers to a cosmetic product used to enhance eyelashes. For example, mascara can darken, thicken, lengthen, and / or define the outline of eyelashes.

[0071] The cosmetic composition may be a mascara composition. The mascara compositions of the present invention are similar to those of currently known mascaras because they incorporate the basic formula ingredients of mascara.

[0072] The mascara composition comprises (i) a plant extract as described herein, or a composition comprising a plant extract as described herein, a water-miscible organic solvent, and optionally water; and (ii) at least one cosmetically acceptable excipient or adjuvant. The at least one cosmetically acceptable excipient or adjuvant comprises at least one wax and optionally at least one pigment. Typically, the mascara composition is an emulsion and comprises a liquid phase (e.g., water) and at least one emulsifier. The mascara composition may also comprise at least one rheology modifier, such as a thickener and / or a film-forming agent.

[0073] As used herein, the term "wax" is intended to refer to a lipophilic aliphatic compound that is solid at room temperature (about 25°C) and atmospheric pressure (760 mmHg, or 105 Pa), undergoes a reversible solid / liquid phase change, and has a melting point exceeding 30°C, exceeding about 55°C in some embodiments, up to about 120°C, or even up to about 200°C. The term "wax" includes waxes of animal origin, waxes of plant origin, waxes of mineral origin, and waxes of synthetic origin. Examples of animal-derived waxes include beeswax and lanolin wax. Examples of plant-derived waxes include rice wax, carnauba wax, candelilla wax, cork fiber wax, sugarcane wax, Japanese wax, sumac wax, cotton wax, and sunflower wax. Examples of mineral-derived waxes include paraffin wax, microcrystalline wax, lignite wax, ceresin wax, and pure ceresin wax. Examples of synthetic waxes include polyolefin waxes, such as polyethylene wax, waxes obtained through Fischer-Tropsch synthesis, wax copolymers and their esters, as well as silicone waxes and fluorinated waxes. Suitable waxes include rice bran wax (Oryza Sativa), Brazilian palm wax (Copernica Cerfifera), and shellac wax. The term "wax" may also include high-melting-point hydrogenated oils of animal or plant origin. Examples include hydrogenated jojoba wax and waxes produced by catalytic hydrogenation from C8-C4. 32 Hydrogenated oils, hydrogenated sunflower oil, hydrogenated castor oil, hydrogenated coconut oil, hydrogenated lanolin, and hydrogenated palm oil are obtained from fats composed of straight-chain or non-straight-chain fatty chains. The mascara composition may contain at least one wax, wherein the total amount of wax in the composition is 0.5% to 20% by weight, or 1% to 10% by weight, or 2% to 8% by weight, or 4% to 7% by weight, based on the total weight of the mascara composition.

[0074] The pigment can be a mineral pigment and / or an organic pigment, and can be coated or uncoated. Among mineral pigments, metal oxides may be mentioned, particularly optionally surface-treated titanium dioxide, zirconium oxide, zinc oxide, or cerium oxide, and iron oxides, particularly black iron oxide, titanium oxide or chromium oxide, manganese violet, ultramarine, chromium hydrate, and iron blue. Among organic pigments, carbon black, D&C type pigments, and lakes based on carmine or barium, strontium, calcium, or aluminum may be mentioned. The mascara composition may contain at least one pigment, wherein the total amount of pigment in the composition is from 1% to 20% by weight, or 5% to 15% by weight, or 8% to 12% by weight, based on the total weight of the mascara composition.

[0075] Emulsifiers suitable for use in mascara compositions include anionic, amphoteric, and nonionic emulsifiers suitable for emulsifying aliphatic compounds in an aqueous phase. In some embodiments of the invention, the emulsifier is selected from fatty acid esters of fatty acids, glycerol, and / or polyalkylene glycols, amphoteric acetates, and alkyl phosphate esters. In some embodiments of the invention, the emulsifier is selected from stearic acid, glyceryl stearate, peg-200 glyceryl stearate, stearyl alcohol polyether-2, stearyl alcohol polyether-20, potassium cetyl phosphate, and disodium cocoamphoacetate. Suitable emulsifiers include Glucate SS, Arlatone MAP160, and mixtures thereof. The mascara composition may contain at least one emulsifier, wherein the total amount of the emulsifier in the composition is 0.01% to 8% by weight, or 0.05% to 8% by weight, or 0.05% to 5% by weight, or 0.05% to 0.5% by weight, or 0.1% to 0.20% by weight, based on the total weight of the mascara composition.

[0076] Mascara compositions may contain rheology modifiers. Rheology modifiers include polymerized natural and derived gums that increase viscosity, resin thickeners, gelling agents, or suspending agents. To increase viscosity, the composition may contain one or more rheology modifiers, which may be synthetic or natural.

[0077] Examples include fatty alcohols, such as C10-C32 alcohols, e.g., C12-C22 alcohols, natural oils, and polymers of acrylic acid and / or methacrylic acid, such as carbomer. Exemplary natural oils include mineral oils (primarily C15-C40 straight-chain and branched aliphatic alkanes with trace amounts of cycloalkanes), naturally derived esters, natural alkanes, or vegetable oils. Exemplary synthetic rheology modifiers comprise acrylic polymers and copolymers. One class of acrylic-based rheology modifiers are carboxyl-functionalized base-swellable and base-soluble thickeners (ASTs) produced by the free radical polymerization of acrylic acid, alone or in combination with other olefinically unsaturated monomers. The polymers can be synthesized using solvent / precipitation and emulsion polymerization techniques. Such exemplary synthetic rheology modifiers include homopolymers of acrylic acid or methacrylic acid, and copolymers polymerized from one or more monomers, including acrylic acid, substituted acrylic acid, and salts of acrylic acid and substituted acrylic acid, and C1-C30 alkyl esters. As defined herein, substituted acrylic acid contains substituents oriented at the α-carbon atom and / or β-carbon atom of the molecule, wherein, in one aspect, the substituents are independently selected from C1-C4 alkyl, -CN, and -COOH. Optionally, other olefinically unsaturated monomers, such as styrene, vinyl acetate, ethylene, butadiene, acrylonitrile, and mixtures thereof, can be copolymerized into the backbone. The aforementioned polymers are optionally crosslinked with monomers containing two or more olefinically unsaturated groups. In one aspect, the crosslinking agent is selected from polyolefin polyethers of polyols containing at least two olefinic ether groups per molecule. Other exemplary crosslinking agents are selected from allyl ethers of sucrose and allyl ethers of pentaerythritol, and mixtures thereof. These polymers are described more fully in U.S. Patents 5,087,445, 4,509,949, and 2,798,053.

[0078] On the one hand, AST rheology modifiers or thickeners are cross-linked homopolymers polymerized from acrylic acid or methacrylic acid and are usually referred to as carbomers under the INCI name. Commercially available carbomers include those from Lubrizol Advanced Materials, Inc. Polymers 934, 940, 941, 956, 980, and 996. In another aspect, the rheology modifier is selected from crosslinked copolymers formed by polymerizing a first monomer and a second monomer, wherein the first monomer is selected from one or more monomers selected from acrylic acid, substituted acrylic acid, salts of acrylic acid, and salts of substituted acrylic acid, and the second monomer is selected from one or more C10-C30 alkyl acrylates selected from acrylic acid or methacrylic acid. In one aspect, these monomers can be polymerized in the presence of a steric stabilizer, such as that disclosed in U.S. Patent No. 5,288,814, which is incorporated herein by reference. Some of the aforementioned polymers are named acrylate / C10-C30 alkyl acrylate crosslinked polymers under INCI nomenclature and may be traded under various names. 1342 and 1382, Ultrez 20 and 21 ETD 2020, and TR-1 and TR-2 were commercially available from Lubrizol Advanced Materials, Inc.

[0079] On the other hand, the rheology modifier may be a crosslinked linear poly(vinylamide / acrylic acid) copolymer as disclosed in U.S. Patent No. 7,205,271, the disclosure of which is incorporated herein by reference.

[0080] Another class of synthetic rheology modifiers suitable for this composition includes hydrophobically modified ASTs, commonly referred to as hydrophobically modified alkali-swellable and alkali-soluble emulsion (HASE) polymers. Typical HASE polymers are free radical addition polymers polymerized from monomers such as: pH-sensitive or hydrophilic monomers (e.g., acrylic acid and / or methacrylic acid), hydrophobic monomers (e.g., C1-C30 alkyl esters of acrylic acid and / or methacrylic acid, acrylonitrile, styrene), "associative monomers," and optional crosslinking monomers. Associative monomers comprise olefinically unsaturated polymerizable end groups and nonionic hydrophilic intermediate segments capped with hydrophobic end groups. Nonionic hydrophilic intermediate segments comprise polyoxyalkylene groups, such as polyethylene oxide, polypropylene oxide, or mixtures of polyethylene oxide / polypropylene oxide segments. The terminal hydrophobic end groups are typically C8-C40 aliphatic moieties. Exemplary aliphatic moieties are selected from straight-chain and branched alkyl substituents, straight-chain and branched alkenyl substituents, carbocyclic substituents, aryl substituents, aralkyl substituents, arylalkyl substituents, and alkylaryl substituents. In one aspect, associative monomers can be prepared by polyethoxylation and / or polypropoxylation of aliphatic alcohols (typically containing branched or unbranched C8-C40 aliphatic moieties) with monomers including: olefinically unsaturated monomers containing carboxylic acid groups (e.g., acrylic acid, methacrylic acid), unsaturated cyclic anhydride monomers (e.g., maleic anhydride, itaconic anhydride, citraconic anhydride), monoolefinically unsaturated monoisocyanates (e.g., α,α-dimethyl-m-isopropenyl isocyanate benzyl ester), or olefinically unsaturated monomers containing hydroxyl groups (e.g., vinyl alcohol, allyl alcohol). Polyethoxylated and / or polypropoxylated aliphatic alcohols are ethylene oxide adducts and / or propylene oxide adducts of monohydric alcohols containing a C8-C40 aliphatic moiety. Non-limiting examples of alcohols containing a C8-C40 aliphatic moiety include: octanol, isooctanol (2-ethylhexanol), nonanol (1-nonanol), decanol, lauryl alcohol, myristol, cetyl alcohol, cetearyl alcohol (a mixture of C16-C18 monohydric alcohols), stearyl alcohol, isostearyl alcohol, transoleyl alcohol, oleyl alcohol, arachidyl alcohol, behenyl alcohol, creosotenic alcohol, montanol, beeswax alcohol, lacceryl alcohol, geddyl alcohol, and C2-C20 alkyl-substituted phenols (e.g., nonylphenol).

[0081] Exemplary HASE polymers are disclosed in U.S. Patents 3,657,175, 4,384,096, 4,464,524, 4,801,671, and 5,292,843. Furthermore, a comprehensive review of HASE polymers can be found in: Gregory D. Shay, Chapter 25, “Alkali-Swellable and Alkali-Soluble Thickener Technology A Review,” Polymers in Aqueous Media—Performance Through Association, Advances in Chemistry Series 223, J. Edward Glass (ed.), ACS, pp. 457-494, Division Polymeric Materials, Washington, DC (1989), the relevant disclosure of which is incorporated herein by reference. Commercially available HASE polymers are marketed under trade names. 22 (INCI name: Acrylate / Stearyl alcohol polyether-20 methacrylate copolymer) 44 (INCI name: PEG-150 / decanol / SMDI copolymer) (INCI name: PEG-150 / stearyl alcohol / SMDI copolymer) and 88 (INCI name: Acrylate / Stearyl alcohol polyether-20 methacrylate crosspolymer) is sold by Rohm & Haas and traded under the name Novethix. TM L-10 (INCI name: acrylate / behenol polyether-25 methacrylate copolymer) is sold by Lubrizol Advanced Materials, Inc.

[0082] In another embodiment, acid-swellable associative polymers can be used as rheology modifiers. Such polymers typically possess cationic and associative properties. These polymers are free-radical addition polymers composed of a mixture of monomers comprising an acid-sensitive amino-substituted hydrophilic monomer (e.g., dialkylaminoalkyl esters of (meth)acrylate or (meth)acrylamide), an associative monomer (as defined above), a lower alkyl ester of (meth)acrylate or other free-radical polymerizable comonomers selected from hydroxyalkyl esters of (meth)acrylate, vinyl and / or allyl ethers of polyethylene glycol, vinyl and / or allyl ethers of polypropylene glycol, vinyl and / or allyl ethers of polyethylene glycol / polypropylene glycol, polyethylene glycol esters of (meth)acrylate, polyethylene glycol esters of (meth)acrylate, polyethylene glycol / polypropylene glycol esters of (meth)acrylate, and combinations thereof. These polymers may optionally be crosslinked. Acid sensitivity refers to the amino substituent becoming cationic at low pH values ​​(typically 0.5 to 6.5). Exemplary acid-swellable associative polymers may be traded under the name... Plus (INCI name: acrylate / aminoacrylate / C10-C30 alkyl PEG-20 itaconic acid ester) was purchased from Akzo Nobel and can be purchased under the trade name. Aqua CC (INCI name: Polyacrylate-1 crosslinked polymer) is commercially available from Lubrizol Advanced Materials, Inc. In one aspect, the acid-swellable polymer is one or more C1-C5 alkyl esters of (meth)acrylic acid, C1-C4 dialkylamino C1-C6 alkyl methacrylates, PEG / PPG-30 / 5 allyl ether, PEG 20-25 methacrylate C10-C30 alkyl ether, or a copolymer of hydroxyl C2-C6 alkyl methacrylate crosslinked with ethylene glycol dimethacrylate. Other useful acid-swellable associative polymers are disclosed in U.S. Patent No. 7,378,479.

[0083] Hydrophobically modified alkoxylated methyl glucosides (such as, respectively, under trade names) DOE-120, Glucamate TM LT, Glucamate TM VLT and Glucamate TM SSE-20 (PEG-120 methyl glucoside, PEG-120 methyl glucoside and PEG-20 methyl glucoside sesquistearate, purchased from Lubrizol Advanced Materials, Inc.) is also suitable as a rheology modifier.

[0084] Polysaccharides obtained from tree and shrub exudates, such as gum arabic, gum gahatti, and tragacanth gum, as well as pectin; seaweed extracts, such as alginate and carrageenan (e.g., λ, κ, ι and their salts); algal extracts, such as agar; microbial polysaccharides, such as xanthan gum, gellan gum, and wellan; cellulose ethers, such as ethylhexylethylcellulose, hydroxybutylmethylcellulose, hydroxyethylmethylcellulose, hydroxypropylmethylcellulose, methylcellulose, carboxymethylcellulose, hydroxyethylcellulose, and hydroxypropylcellulose; polygalactomannans, such as fenugreek gum, cinnamon gum, locust bean gum, tara gum, and guar gum; and starches, such as corn starch, cassava starch, rice starch, wheat starch, potato starch, and sorghum starch, may also be used as suitable rheology modifiers in the compositions herein. A particularly suitable rheology modifier as a thickener is Diutan gum.

[0085] The rheology modifier can be used alone or in combination, and can be present in the composition at a total concentration of 0.001% to 50% by weight, for example, at least 0.1% by weight, or at least 1% by weight, such as up to 20% by weight, or up to 10% by weight, or up to 3% by weight, depending on the active ingredient and based on the total weight of the composition.

[0086] Mascara compositions may contain a rheology modifier as a film-forming agent. The film-forming agent is dissolved in at least one solvent (such as water and / or an organic solvent) in the mascara composition, and after the mascara composition is applied to hair, at least one solvent evaporates, is absorbed, and / or dissipates on the hair, leaving a film on the hair. The use of a film-forming agent improves the abrasion resistance of the mascara and can impart anti-transfer properties. Film-forming agents are well known in the art and can be any cosmetically acceptable film-forming agent used around the eyes. Examples of useful film-forming agents include natural waxes, polymers such as acrylic copolymers, polyethylene polymers, and copolymers of polyvinylpyrrolidone (PVP), ethylene-vinyl acetate, and polydimethylsiloxane gum, as well as resins such as shellac, polyterpenes, and various silicone resins, such as trimethylsiloxysilicate. Film-forming agents can be PVP, acrylic copolymers, and polyurethanes. Suitable film-forming agents are polyurethanes, such as, but not limited to, Avalure. TM UR 450 polymer. The film-forming agent can be used in amounts from about 0.1% to about 50% by weight, or from about 0.5% to about 20.0% by weight, based on the total weight of the mascara composition.

[0087] In addition, these cosmetic compositions may contain other active ingredients such as vitamins, minerals, proteins, peptides, fatty acids, antioxidants, anti-inflammatory agents, color enhancers, and / or mixtures thereof. In particular, cosmetic compositions may contain other active ingredients for promoting hair growth and / or preventing hair loss. In the context of this invention, a suitable non-limiting example is Growth, sold by Hallstar. [INCI: Helianthus Annuus (sunflower) seed oil (and) polyglyceryl-3 diisostearate (and) Carthamus Tinctorius (safflower) flower extract (and) Hibiscus Sabdariffa (hibiscus) flower extract]. Widelash sold by Sederma TM [INCI: Glycerin, Water (Aqua), Panthenol, Biotin Tripeptide-1]; Sold by Symrise Xlash [INCI: glycerol (and) Aqua (and) myristoyl pentapeptide-17], 1631 [INCI: Pentylene glycol (and) Isochrysis Galbana extract]; Anargy, sold by Lipotrue TM [INCI: Water (and) Butylene Glycol (and) Oligopeptide-2 (and) Nicotiana Benthamiana Hexapeptide-40SH-Polypeptide-9 (and) Nicotiana Benthamiana Hexapeptide-40SH-Polypeptide-86]; Capixyl sold by Lucas Meyer TM [INCI: Butylene Glycol (and) Aqua (and) Dextran (and) Acetyl Tetrapeptide-3 (and) Trifolium Pratense (Alfalfa) Flower Extract]; Nano lashes sold by Nanovetores [INCI: Aqua, Simmondsia Chinensis Seed Extract, Polysorbate 20, Hydroxypropyl Guar Gum, Sodium Benzoate, Potassium Sorbate]; Specped SC-MH16 sold by Spec-chem [INCI: Aqua, Glycerin, Myristoyl Hexapeptide-16, Caprylyl Glycol, Ethylhexylglycerin], Myristoyl Pentapeptide-17 [INCI: Myristoyl Pentapeptide-17] BT1 [INCI: Biotinylate Tripeptide-1], LashLD [INCI: Biotin tripeptide-1, glycerol, water, capryloyl glycol and ethylhexylglycerol, panthenol, trehalose], MP17P [INCI: Myristoyl Pentapeptide-17]; Procapil sold by SedermaTM [INCI: Butylene Glycol (and) Aqua (and) PPG-26-Buteth-26 (and) PEG-40 Hydrogenated Castor Oil (and) Apigenin (and) Oleanolic Acid (and) Biotin Tripeptide-1]; AnaGain sold by Mibelle TM [INCI: Pisum Sativum (pea) sprout extract (and) isomaltitol (and) Aqua], PhytoCellTec TM Malus DomesticaHair [INCI: Malus Domestica Fruit Cell Culture Extract (and) Xanthan Gum (and) Glycerin (and) Lecithin (and) Phenoxyethanol (and) Aqua / Water], Santenergy TM [INCI: Bioflavonoids (and) Pentylene glycol (and) Alcohol (and) Aqua], RootBioTec TM HW [INCI: Ocimum Basilicum hairy root culture extract (and) alcohol (and) Aqua / water], RootBioTec TM HO [INCI: Basil hairy root culture extract (and) Helianthus Annuus (sunflower) seed oil (and) Cocos Nucifera (coconut) oil]; CAPILIA LONGA PPF [INCI: Curcuma Longa (turmeric) callus culture conditioned medium (and) water] sold by Vytrus Biotech; sold by Solabia [INCI: Glycerin (and) Aqua (and) Watercress Extract (and) Nasturtium Extract]; Alotide sold by Peptron TM [INCI: Copper Ascorbate Succinyl Tripeptide-34]; BURGEON-UP [INCI: Aqua(and) Alcohol(and) Watercress Leaf / Stem Extract] sold by Ichimaru Pharcos; sold by Exysmol [INCI: Chondrus Crispus Extract]; sold by BASF GY[INCI: Aqua (and) glycerol (and) glycogen (and) phenoxyethanol (and) methylparaben], Trichigen TMVEG LS 9922 [INCI: Water (and) Panax Ginseng Root Extract (and) Arginine (and) Acetyltyrosine (and) Arctium Majus Root Extract (and) Hydrolyzed Soy Protein (and) Polyquaternium-11 (and) PEG-12 Polydimethylsiloxane (and) Calcium Pantothenate (and) Zinc Gluconate (and) Nicotinamide (and) Ornithine Hydrochloride (and) Citrulline (and) Glucosamine Hydrochloride (and) Biotin]; Follicusan sold by CLR TM DP [INCI: Aqua (and) denatured alcohol (and) ubiquitin ethyl ether (and) inositol (and) milk protein (and) lactose (and) acetylcysteine ​​(and) acetylmethionine (and) sodium citrate (and) citric acid]; sold by Greentech [INCI: Propylene Glycol (and) Water (Aqua) (and) Lindera Strychnifolia Root Extract]; Kerascalp sold by Provital TM [INCI: Propylene Glycol (and) Glycerin (and) Phyllanthus Emblica Fruit Extract], Baicapil TM [INCI: Propylene Glycol (and) Water (and) Arginine (and) Lactic Acid (and) Wild Soybean (Glycine Soja) Germ Extract (and) Triticum Vulgare (Wheat) Germ Extract (and) Scutellaria Baicalensis Root Extract (and) Sodium Benzoate (and) Gluconolactone (and) Calcium Gluconate]; BIOENERGIZER sold by Seppic TM P BG PF [INCI: Aqua / Water, Butylene Glycol, Panthenol, Propylene Glycol, Pelvetia Canaliculata Extract, Laminaria Digitata Extract]; Protectagen sold by Ashland TM [INCI: Aqua (and) glycerol (and) hydrolyzed rice protein]; sold by Givaudan [INCI: Glycerin (and) Aqua (and) Sodium metabisulfite (and) Larch (Larix Europaea) wood extract (and) Glycine (and) Zinc chloride (and) Camellia Sinensis leaf extract]; sold by B&G [INCI: Glycerin (and) Water (and) Ganoderma Lucidum (mushroom) mycelium fermentation filtrate]; sold by Silab [INCI: Saccharomyces cerevisiae extract] [INCI: Hydrolyzed Lupin Protein]; Hairdian AP [INCI: Propylene Glycol (and) Thuja Orientalis Extract (and) Zingiber Officinale (ginger) Root Extract (and) Red Alfalfa (red clover) Leaf Extract (and) Artemisia Argyi (artemisia argyi) Leaf Extract], sold by Shanghai GREAF Biotech.

[0088] The cosmetic composition may also contain ingredients selected from the group consisting of: (Luo Jian) (Propecia) and (Avodat).

[0089] application

[0090] This invention is based on the discovery that plant extracts from species of the genus *Flavorum* possess some surprising properties. Specifically, these properties enable the use of plant extracts in non-therapeutic cosmetic applications. The purpose of non-therapeutic cosmetic applications is to improve or maintain an aesthetic appearance; in particular, this invention relates to the use of plant extracts to improve or maintain the aesthetic appearance of hair, especially the aesthetic appearance of eyelashes and / or eyebrows.

[0091] It has been found that plant extracts from the genus *Flavorum* can stimulate hair follicle cell proliferation, thereby stimulating new hair formation and promoting hair growth. Furthermore, these extracts have been found to stimulate angiogenesis in endothelial cells and stimulate keratinocyte proliferation, thereby promoting hair growth and increasing hair strength. In addition, it has been found that these extracts increase the expression of type XVII collagen and α6β4 integrin in hair follicles, thereby increasing hair adhesion. Therefore, the plant extracts of the present invention can promote hair growth, prevent hair loss, and / or increase hair thickness.

[0092] This invention provides the use of plant extracts from species of the genus *Flavorum* for promoting hair growth. The term "promoting hair growth" means stimulating or enhancing hair growth, and refers to increasing the density of hair growing in a given area of ​​the human body (i.e., per cm²). 2 (Number of hairs on the skin). Optionally or otherwise, the term also refers to an increase in the length of hair fibers, that is, an increase in the length of hair in a given area over a given period of time compared to the hair length in a given area without the use of extracts from *Flavorum* species over the same period of time.

[0093] When extracts of *Flavorophyllum* species are used, hair density can be increased by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 100%, or at least 200% compared to hair density before the use of extracts of *Flavorophyllum* species.

[0094] When extracts of *Flavorum* species are used, hair length can be increased by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 100%, or at least 200% compared to hair length (where average length) when extracts of *Flavorum* species are not used during the same time period.

[0095] This invention provides the use of plant extracts from species of the genus *Flavorum* for promoting the growth of eyelashes and eyebrows.

[0096] This invention provides the use of plant extracts from species of the genus *Flavorum* for lengthening eyelashes.

[0097] This invention provides the use of plant extracts from species of the genus *Flavorum* for increasing the density of eyelashes.

[0098] This invention provides the use of plant extracts from species of the genus *Flavorum* for lengthening eyebrow hairs.

[0099] This invention provides the use of plant extracts from species of the genus *Flavorum* for increasing the density of eyebrow hairs.

[0100] This invention provides the use of plant extracts from species of the genus *Flavorum* for preventing hair loss. Hair loss can be normal daily hair loss or hair loss due to aging. Hair loss may be caused by mechanical stress, such as mechanical stress caused by hair styling, or, especially in the case of eyelashes and eyebrows, may be caused by rubbing with hands or fingers. In particular, this invention provides the use of plant extracts from species of the genus *Flavorum* for preventing hair loss in eyelashes and eyebrows.

[0101] As used herein, the term "prevent" refers to the ability of plant extracts from species of the genus *Pterocarya* to prevent, delay, or inhibit hair loss. The term "prevent" is used interchangeably with the term "reduce," meaning that the term refers to the ability of plant extracts to reduce the amount of hair loss.

[0102] This invention provides the use of plant extracts from species of the genus *Flavorum* for increasing hair adhesion. For example, this makes the hair more resilient to mechanical stress.

[0103] This invention provides the use of plant extracts from species of the genus *Flavorum* for increasing hair thickness. The term "thickness," when used to describe hair, refers to the average diameter or cross-sectional area of ​​a hair strand. In some embodiments, when using *Flavorum* extracts, hair thickness increases by at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 100%, or at least 200% compared to hair thickness before using the *Flavorum* extract.

[0104] It has been confirmed that plant extracts from species of the genus *Flavorum* can induce the proliferation of dermal papilla cells and keratinocytes in hair follicles, increase blood flow in hair follicles, and increase the synthesis of XVII collagen and α6β4 integrin in hair follicles. Therefore, this invention also relates to the use of plant extracts from species of the genus *Flavorum* to promote hair growth and / or prevent hair loss and / or increase hair thickness by: inducing the proliferation of dermal papilla cells in hair follicles; inducing the proliferation of keratinocytes and / or associated keratinocytes to form hair; increasing blood flow in hair follicles; and increasing the synthesis of XVII collagen and α6β4 integrin in hair follicles.

[0105] In one aspect, the present invention provides a method for promoting hair growth and / or preventing hair loss and / or increasing hair thickness, comprising applying an extract of at least one plant from a species of the genus *Pterocarya* as an active ingredient to a subject. The subject is preferably a mammal, more preferably a human. Preferably, the method is a non-therapeutic cosmetic procedure.

[0106] In the above uses and methods, plant extracts may be included in the compositions described above.

[0107] Plant extracts are preferably applied topically. As used herein, “topical application” means contacting a plant extract (or a composition containing a plant extract) with mammalian keratinized tissue, such as hairy skin. In particular, skin is hairy skin, such as the scalp, facial skin such as mustache or beard, eyebrows, and eyelash area. More particularly, the extract or composition is applied to the eyelash line and / or eyebrows, preferably to the eyelash line.

[0108] The frequency of application or administration of the method of the present invention described above may vary greatly depending on the needs of each subject. The recommended application frequency is once a month to ten times a day, preferably once a week to four times a day, more preferably three times a week to twice a day, or even more preferably once a day.

[0109] The invention also extends to combinations of the method according to the invention with other methods for promoting and / or increasing hair growth, and / or preventing hair loss, and / or increasing hair thickness.

[0110] The improvement or maintenance of hair aesthetics provided by the above uses and methods can be applied to healthy subjects, as well as those with diseases and / or conditions of the skin, hair, nails, and / or mucous membranes. Therefore, the methods of the present invention can be applied to subjects who do not suffer from hair loss, or those who suffer from menopausal hair loss, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium, or cicatricial alopecia. However, the use of the present invention is limited to subjects who do not suffer from hair loss, or those who suffer from menopausal hair loss, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium, or cicatricial alopecia.

[0111] On the other hand, an extract of a species of *Fragaria* plant is provided for use as a medicine. Specifically, an extract of a species of *Fragaria* plant is provided for treating conditions associated with hair loss or thinning hair, wherein such conditions include, but are not limited to, alopecia (including menopausal alopecia, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium, and cicatricial alopecia). On the other hand, a method of treating conditions associated with hair loss or thinning hair, wherein such conditions include, but are not limited to, alopecia (including menopausal alopecia, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium, and cicatricial alopecia), is provided, the method comprising administering a therapeutically effective amount of the extract of a species of *Fragaria* plant to a subject.

[0112] The invention is further described through the following non-limiting embodiments.

[0113] Example 1

[0114] Extracts obtained through conventional aqueous extraction

[0115] Routine extraction was performed on dried, leafy stems of *Myrothamnus f. labellifolia*, cut into segments of approximately 1.27 mm to 0.22 mm. The plant material was extracted with water at a ratio of 1:10 (dried plant material: water, w / v). Extraction was carried out with water at 45°C for 4 hours with stirring. The resulting solution was cooled to room temperature (25°C) and filtered through a 10 μm filter. The filtered solution was the plant extract, which was then diluted with 100% glycerol at a ratio of 1:5 (extract: glycerol, w / w), and the resulting composition was stirred to form a stock solution.

[0116] Example 2

[0117] Extract obtained by subcritical water extraction

[0118] Subcritical water extraction (SWE) was performed on dried, leafy Myrothamnus fruticosa stems ground to a coarse powder of approximately 1.27 mm to 0.25 mm. The powder was extracted with subcritical water at a ratio of 1:20 (powder:water, w / v). The subcritical water was maintained at 150 °C for 1 hour under a pressure ranging from 10 MPa to 11 MPa. Subsequently, the resulting solution was cooled to room temperature (25 °C) and filtered through a 10 μm filter. The filtered solution was the plant extract, which was then diluted with glycerol at a ratio of 1:5 (extract:glycerol, w / w). The resulting composition was stirred to form a stock solution.

[0119] Example 3

[0120] Compositional analysis of extracts

[0121] The differences in phytochemicals in the stock solutions of Examples 1 and 2 were evaluated. The total polyphenol content (TPC) was analyzed, and various phytochemicals were identified.

[0122] The total polyphenol content was determined by the Folin-Ciocalteu method. In short, 2 mL of the extract was mixed with 5 mL of distilled water, then 1 mL of Folin-Ciocalteu reagent was added to the mixture, and the mixture was thoroughly mixed for 3 minutes. After 3 minutes, 5 mL of 10% sodium carbonate solution was added to stop the reaction. After 30 minutes at room temperature, the absorbance at 760 nm and 850 nm was measured using a spectrophotometer. Gallic acid was used as a standard to obtain a calibration curve.

[0123] More specific phytochemicals were identified using liquid chromatography-mass spectrometry (LC / MS-MS). Ten microliters (10 μL) of extract were mixed with 100 μL of 10 mM HCl and 890 μL of ultrapure water in a 2 mL LC-MS vial, diluting the sample 100-fold. The injection volume was 2 μL. The mobile phase used for this analysis consisted of an acetonitrile solution of 0.1% formic acid (solvent A) and an ultrapure aqueous solution of 0.1% formic acid (solvent B). The flow rate was 0.2 mL / min at a column temperature of 40 °C.

[0124] The total polyphenol content of the aqueous plant extract of Example 1 was determined to be 652 mg GAE / L by the Folin-Ciocalteu method, while the total polyphenol content of the stock solution of Example 2 was 763 mg GAE / L. All compounds identified in the stock solutions of Examples 1 and 2 are listed in Table 1. ND markings indicate compounds not detected in the samples.

[0125]

[0126]

[0127] Table 1

[0128] Example 4

[0129] In vitro proliferation assay of dermal papillary cells of hair follicles

[0130] The Myrothamnus flabellifolia stock solution (“Test Product”) obtained as described in Examples 1 and 2 was dissolved in papillary cell growth medium supplemented with growth supplements at concentrations of 0.5% (v / v), 1% (v / v), 2% (v / v), 2.5% (v / v) and 5% (v / v) and filtered through a 0.2 μm syringe filter.

[0131] Hair follicle dermal papillary cells (HFDPCs) from adult scalp were placed in papillary cell growth medium with 5% CO2 humidified air and grown at 37°C until they reached a density of 3 × 10⁻⁶. 4 A density of 100 cells / mL was maintained for 5 days. Subsequently, the cells were separated and then inoculated for treatment.

[0132] HFDPCs were seeded at a density of 45,000 cells / mL in 96-well Cell Carrier black plates (PerkinElmer). After incubation at 37°C in humidified air with 5% CO2 for 24 hours, the medium was removed, and fresh medium and dilutions of the test product and media controls were added. Cells treated with the medium alone served as a negative control. Fetal bovine serum (FBS) was used as a positive control.

[0133] After incubation for 24 hours, use The Cell Viability / Cytotoxicity Kit (from ThermoFisher Scientific) quantifies live and dead cells by simultaneously staining to distinguish between them.

[0134] The test product was measured in three independent experiments, with each experiment including four replicates.

[0135] Table 2 shows the percentages of live and dead HFDPCs normalized to the total cell count (ALL) in the negative control according to the following formula:

[0136]

[0137] Table 2 also shows the proliferation based on the following formula:

[0138]

[0139]

[0140]

[0141] Table 2

[0142] The results showed that, compared with the negative control (untreated cells), Myrothamnus flabellifolia extract increased the number of live cells and reduced the number of dead cells. Subsequently, these extracts had a proliferative effect on HFDPCs.

[0143] Example 5

[0144] In vitro proliferation assay of keratinocytes

[0145] Keratinocytes express keratin (the most abundant protein in hair fibers) and help hair to form from the top.

[0146] Myrothamnus flabellifolia stock solution was dissolved at 0.5% (v / v), 1% (v / v), 2% (v / v), 2.5% (v / v), and 5% (v / v) in keratinocyte growth medium supplemented with Mix C39016-CaCl2 solution and filtered through a 0.2 μm syringe filter.

[0147] Human epidermal keratinocytes (HEK) were placed in a keratinocyte growth medium humidified with 5% CO2 air and grown to 4500 cells / cm³ over 7 days at 37°C. 2 The density. Afterwards, the cells are separated and then inoculated for treatment.

[0148] HEK cells were seeded at a density of 70,000 cells / mL in 96-well Cell Carrier black plates (PerkinElmer). After incubation at 37°C with 5% CO2 humidified air for 24 hours, the medium was removed, and fresh medium and dilution of the test product were added. Cells treated with medium only served as a negative control. Epidermal growth factor (EGF) served as a positive control. After 24 hours of incubation, [the cells were then]... Cell viability reagents are used to quantify live and dead cells.

[0149] Using a cell-permeable rezindra-based solution from Thermo Fisher Scientific Fluorescence quantification was performed to quantify HEK.

[0150] The stock solutions of Examples 1 and 2 were determined in three independent experiments, each of which included four replicates.

[0151] result

[0152] Table 3 shows the cell growth values ​​of HEK cells normalized relative to the negative control.

[0153]

[0154]

[0155] Table 3

[0156] The results showed that Myrothamnus flabellifolia extract increased the proliferation of keratinocytes.

[0157] Example 6

[0158] In vitro study of angiogenesis in human umbilical vein endothelial cells (HUVECs)

[0159] Hair follicles receive nutrition from the blood flow in the skin where the hair is attached. Optimal irrigation of hair follicles increases their nutrition and stimulates new hair growth, while also improving hair growth and increasing hair strength. Compounds capable of generating new blood vessels in hair follicles are promising candidates for use in eyelash cosmetic care to promote eyelash growth and increase eyelash strength. In vitro assessment of angiogenesis involves measuring the ability of endothelial cells to form three-dimensional tubular structures. Human umbilical vein endothelial cells (HUVECs) mediate specific connections with extracellular matrix components, generating differential forces that induce morphological changes and migration of HUVECs, thereby causing tubular structure formation. The aim of this study is to evaluate the efficacy of the compounds of this invention in generating new blood vessels by measuring the ability of endothelial cells to form three-dimensional tubular structures.

[0160] HUVEC cells were pre-coated in 96-well plates with cold extracellular matrix solution (PromoKine angiogenesis assay kit) and incubated at 37°C for 1 hour. After incubation, cells were seeded at a density of 15,000 cells / well. Cells were immediately treated with the stock solution from Example 2 dissolved in culture medium to achieve a final well concentration of 0.5% (v / v). Vascular endothelial growth factor (VEGF) was used as a positive control at a final well concentration of 5 ng / ml, and cells treated with culture medium alone served as a basal control. Cells were treated in 5% CO2 humidified air at 37°C for 4 hours, and then stained with the staining dye (PromoKine angiogenesis assay kit) according to the manufacturer's instructions. Briefly, cells were washed and then the staining dye solution was added. After incubation in 5% CO2 humidified air at 37°C for 30 minutes, HUVEC cells were seeded at a density of 15,000 cells / well. Fluorescence images were captured using confocal microscopy (PerkinElmer, Inc.), and new blood vessel formation was evaluated using ImageJ software.

[0161] Angiogenesis is evaluated by measuring different parameters in each acquired image. First, ImageJ detects the network's constituent elements, such as connectors, segments, and branches. Connectors are nodes that correspond to vessel bifurcation, segments are vessels defined by two connectors, and branches correspond to elements defined by only one connector. The software then quantifies more complex structures:

[0162] - Line segment length: The number of pixels for each detected line segment.

[0163] - Number of main segments: The number of segments defined by connection points without branches.

[0164] - Number of main connection points: The number of connection points that link at least three main line segments.

[0165] Grid number: The number of elements defined by line segments or main line segments.

[0166] Given the increased formation of new blood vessels, it is necessary to increase the value of more than one of these measurement parameters.

[0167] The results in this paper are expressed as percentages of different measurement parameters normalized by the baseline conditions.

[0168]

[0169] Table 4

[0170] The percentage of different measurement parameters for HUVEC cells relative to the mean of baseline conditions.

[0171] The results confirmed that the plant extract of the present invention, at the tested concentration, increased tubular formation relative to basal conditions in HUVEC cells. The results also demonstrated that the increase achieved by the plant extract of the present invention was similar to that achieved by the positive control.

[0172] Example 7

[0173] In vitro study of immunostaining of type XVII collagen and integrin β4 in scalp explants

[0174] Hair attachment depends on the adhesion of different cells to the basement membrane region, with hemidesmosomes being the main binding unit. Hemidesmosomes are cell-matrix junctions formed by type XVII collagen, two subunits of α6β4 integrin, and CD151. Both type XVII collagen and α6β4 integrin interact directly with the basement membrane region, thus helping to prevent hair loss. In addition, the degradation of type XVII collagen is associated with hair follicle aging. It has been shown that during aging, hair follicles miniaturize and often disappear from the skin via a miniaturization process, characterized by the degradation of type XVII collagen, leading to thinning and loss of hair [Matsumura H, et al. "Hair follicle aging is driven by transepidermalelimination of stem cells via COL17A1 proteolysis" Science. 2016 Feb 5; 351(6273):aad4395]. Compounds capable of increasing type XVII collagen and α6β4 integrin are promising candidates for eyelash and eyebrow cosmetic care to combat aging, increase hair strength, and prevent hair loss. The aim of this study was to evaluate the efficacy of the compounds extracted in this invention in improving hair fixation by measuring increases in type XVII collagen and integrin β4 (β4) in different regions of hair follicles from scalp explants.

[0175] Scalp explants were prepared and stored in BEM medium (BIO-EC) at 37°C for 8 days under a humid 5% CO2 atmosphere. On days 0, 4, and 6, a 2% (w / w) dilution of the stock solution from Example 2 was locally applied to the explants. Scalp explants treated only with the culture medium served as a basal control. On day 8, the explants were collected and cut into two halves. One half was fixed in formalin buffer, and the other half was frozen at -80°C.

[0176] After fixation in buffered formalin for 24 hours, the explants were dehydrated and then impregnated with paraffin. The explants were then embedded, and 5 μm thick sections were prepared using a microtome (Leica). The sections were mounted on histological slides. Using the Vectastain kit vector amplification system with avidin / biotin, type XVII collagen was immunostained with monoclonal anti-XVII collagen antibody (Abcam) at room temperature for 1 hour, followed by staining with VIP (Vector laboratories, Ref. SK-4600). VIP is a peroxidase substrate that stains purple upon oxidation. Immunostaining was evaluated by microscopic observation, followed by semi-quantitative analysis using image analysis.

[0177] Frozen explants were sectioned into 7 μm thick slices using a cryostat (Leica). The slices were then mounted on silanized slides. Frozen skin sections were immunostained with monoclonal anti-integrin β4 antibody (Chemicon) at room temperature for 1 hour, followed by development using Alexa Fluor 488 (Life Technologies). Cell nuclei were counterstained with propidium iodide. Immunostaining was performed manually, followed by semi-quantitative analysis using image analysis.

[0178] Microscopic observations of type XVII collagen and integrin β4 were imaged using an Olympus camera and then analyzed using CellSens software. Different regions of the hair follicle were studied by defining the following regions of interest: the dermal-epidermal junction, the infundibulum, the upper root sheath, the ridge, the lower root sheath, and the hair bulb. In each of these regions, type XVII collagen and integrin β4 were quantified.

[0179] The results of this study show the percentage increase in type XVII collagen and integrin β4 in the hair follicle region relative to the baseline control.

[0180] dermal-epidermal junction 28.25％ <![CDATA[55.98% *** ]]> Funnel section <![CDATA[57.70% * ]]> <![CDATA[187.76% **** ]]> upper root sheath - <![CDATA[209.35% **** ]]> Lontu District - <![CDATA[60.20% *** ]]> inferior root sheath <![CDATA[31.40% * ]]> <![CDATA[101.11% ** ]]> fluff ball <![CDATA[255.97% * ]]> <![CDATA[107.18% * ]]>

[0181] Table 5

[0182] The percentage increase in type XVII collagen and integrin β4 in different regions of the hair follicle relative to the baseline control. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001)

[0183] The results demonstrate that the plant extract of the present invention stimulates hair fixation by increasing the expression of type XVII collagen and integrin β4 along the hair follicle.

[0184] Example 8

[0185] eyelash mascara

[0186] The eyelash mascara according to the present invention is prepared as follows:

[0187] Phase A components were mixed with stirring while being heated at 85°C, and then dispersed into Phase B components. Subsequently, Phase D components were added.

[0188] The components of phase E are mixed while heating at 90°C. Phase E is slowly added to the mixture of phases A through D, and phase F is added last to prepare the emulsion.

[0189]

[0190]

[0191] Table 6

[0192] Example 9

[0193] eyelash serum

[0194] Mix the components of phase A together. Then add the components from phase B while stirring. Neutralize the mixture with the components of phase C, and finally add component D.

[0195]

[0196] Table 7

[0197] Example 10

[0198] This study was used to evaluate the increase in eyelash length following long-term application of extracts from *Pteris vittata* species in Caucasian female volunteers. In vivo studies in Canada .

[0199] This study lasted 56 days. The volunteer group consisted of thirty-one (31) white female volunteers aged between 25 and 50 years. All individuals in this study were given the active and placebo products under a “half-face” design, with the active serum of Example 9 and the mascara of Example 8 applied to the lashes on one half of the face, and the placebo serum and mascara (with the same formula but without the *Pyrrosia* species extract) applied to the lashes on the other half of the face. The serum was applied to the lash roots in the morning and evening, and the mascara was applied only in the morning for 56 days. During the treatment, all volunteers used Beiesdorf's facial cleanser only at night. DermatoCLEAN washes off the morning serum and mascara, followed by the evening serum application. Subjects served as self-references, comparing results obtained at 56 days with those obtained at the initial time. Furthermore, results obtained with the active formulation were compared to those obtained with the placebo formulation. The efficacy of the composition was assessed by measuring eyelash length.

[0200] Before and 56 days after product use, lateral macroscopic photographs taken at a 20° angle to the face were taken using CameraScan (Orion Concept) software. The eyes were closed, ensuring the upper eyelid eyelashes were positioned on a white patch. Images were acquired in parallel polarized light and analyzed using ImageJ (National Institutes of Health) software. The imaged eyelashes were divided into five sections, and the longest eyelash in each section was selected for measurement. The results are shown in Table 8.

[0201]

[0202] Table 8

[0203] After 56 days of product application, the average eyelash length increased. Statistical significance relative to the initial time: *p<0.05, calculated using an unpaired t-test relative to the initial time.

[0204] Results showed that, after 56 days of application of the serum and mascara of this invention, the average eyelash length increased statistically significantly compared to the initial time. Furthermore, the increase in eyelash length was greater when the active formulation was used compared to the placebo formulation.

[0205] Example 12

[0206] This study was used to evaluate the improvement in eyebrow density following long-term application of a styrax extract in Caucasian female volunteers. In vivo studies .

[0207] This study lasted 56 days. The volunteer group consisted of twenty-eight (28) Caucasian female volunteers aged between 25 and 50 years. All individuals in this study received both the active serum of Example 9 and a placebo serum (with identical ingredients but without the *Flavorum* species extract) in a “half-face” design. The serum was applied to the eyebrows twice daily (morning and evening) during the 56 days of the study. In addition, all volunteers used Beiesdorf's facial cleanser only at night during the treatment period. DermatoCLEAN washes off the morning-applied serum and then applies the serum at night. Subjects act as their own references, comparing results obtained at 56 days with results obtained at the initial time. Furthermore, results obtained with the active serum are compared with results obtained with the placebo serum.

[0208] The efficacy of the composition was evaluated by measuring eyebrow density. Lateral macroscopic photographs taken at a 35° angle to the face were taken using CameraScan (Orion Concept) software before and 56 days after product use. The obtained images were analyzed using ImageJ software. The eyebrow area was selected and converted into a binary image. The white pixel count was performed, and the following results are displayed:

[0209]

[0210] Table 9

[0211] After 56 days of product application, the average eyebrow density increased. Statistical significance relative to the initial time: ls p < 0.1, calculated using an unpaired t-test relative to the initial time.

[0212] Results showed that after 56 days of applying the essence of this invention, eyebrow density increased compared to the initial time.

[0213] Example 13

[0214] Used to evaluate eyelash curvature angle after long-term application of extracts from *Flavorum* species in Caucasian female volunteers. In vivo studies .

[0215] This study lasted 56 days. The volunteer group in this study consisted of thirty-one (31) white female volunteers aged between 25 and 50 years. All individuals in this study were given the active product and placebo products under a “half-face” design, with the active serum and mascara applied to the lashes on one half of the face and the placebo serum and mascara applied to the lashes on the other half of the face. The serum was applied to the lash roots once in the morning and once at night; mascara was applied only in the morning for 56 days. During the treatment, all volunteers used a facial cleanser (from Beiesdorf) only at night. DermatoCLEAN washes off the morning-applied serum and mascara, followed by the evening application of the serum. The active mascara and serum are described in Examples 8 and 9, respectively. The placebo mascara and serum have the same composition as the active mascara and serum, except that they lack the *Pterocarya* species extract. Subjects served as their own references, and results obtained at 56 days were compared to results obtained at the initial time. The efficacy of the composition was assessed by determining changes in eyelash curvature angle using the following formula:

[0216]

[0217] Average parameters of all volunteers after x days of product use

[0218] The average parameter values ​​of all volunteers at baseline

[0219] A reduction in eyelash curvature is considered a beneficial effect.

[0220] Before product use (at the initial time) and 56 days after product use, lateral macroscopic photographs at a 90° angle to the face were taken using CameraScan (OrionConcept) software. Images were acquired in parallel polarized light and analyzed using ImageJ (National Institutes of Health) software. The uppermost eyelash in each image was selected, and the curvature angle was measured at the midpoint of that eyelash.

[0221]

[0222] Table 10

[0223] Changes in average eyelash curvature angle 56 days after product application. Statistical significance relative to the initial time:

[0224] ***p<0.001, calculated using an unpaired Student's t-test relative to the initial time.

[0225] The results showed that, after 56 days of applying the essence and mascara of the present invention, the average eyelash curvature angle was significantly reduced compared to the initial time.

[0226] Example 14

[0227] In vivo studies used to assess hair loss in eyelashes and eyebrows

[0228] This study lasted 28 days. The study included thirty-one (31) white female volunteers aged between 25 and 50 years. All individuals in this study were given both the active and placebo products using a “half-face” design. For eyelashes, the active serum and mascara were applied to the eyelashes on one half of the face, and the placebo serum and mascara were applied to the eyelashes on the other half of the face. The serum was applied to the lash roots once in the morning and once at night, with mascara applied only in the morning for 28 days. Eyebrows were treated with the same active and placebo half-design. The serum was applied to the eyebrows both in the morning and at night for 28 days. During the treatment, all volunteers washed off the morning-applied serum and mascara only at night with a facial cleanser, after which the serum was applied again at night. The active serum and mascara are described in Examples 8 and 9. Results obtained with the active formulation were compared with results obtained with the placebo formulation. The efficacy of the compositions was evaluated by the total number of hairs shed from the eyelashes and eyebrows.

[0229] Hair loss was counted daily for eyelashes and eyebrows, and each eyelash and eyebrow was then cleaned separately to count hair loss when using the active product and the placebo product. The results shown in Table 11 correspond to the total number of hair losses for all volunteers for each condition and location.

[0230]

[0231] Table 11. Total number of hairs lost from eyelashes and eyebrows when using active and placebo products.

[0232] Results showed that, 28 days after applying the essence and mascara of the present invention, the number of hairs falling out of eyelashes and eyebrows was reduced when the active care products were applied, compared with the application of a placebo.

Claims

1. A plant extract of a species of the genus *Fragaria* for non-therapeutic cosmetic purposes of promoting hair growth and / or preventing hair loss and / or increasing hair thickness, wherein the plant extract is a plant extract of the species *Myrothamnus flabellifolia*, and wherein the extract is obtained by solid / liquid extraction using subcritical water as the extraction solvent.

2. A plant extract of a species of the genus *Fragaria* for non-therapeutic cosmetic use to lengthen eyelashes, wherein the plant extract is a plant extract of the species *Myrothamnus flabellifolia*, and wherein the extract is obtained by solid / liquid extraction using subcritical water as the extraction solvent.

3. A plant extract of a species of the genus *Fragaria* for non-therapeutic cosmetic use to increase the density of eyebrow hairs, wherein the plant extract is a plant extract of the species *Myrothamnus flabellifolia*, and wherein the extract is obtained by solid / liquid extraction using subcritical water as the extraction solvent.

4. Plant extracts of species of the genus *Flavorum* for non-therapeutic cosmetic purposes of increasing hair adhesion, wherein the plant extract is a plant extract of the species *Myrothamnus flabellifolia*, and wherein the extract is obtained by solid / liquid extraction using subcritical water as the extraction solvent.

5. The non-therapeutic cosmetic use according to claim 1 or claim 4, wherein the hair is eyelash or eyebrow hair.

6. A non-therapeutic cosmetic method for promoting hair growth and / or preventing hair loss and / or increasing hair thickness, the method comprising applying an extract of at least one plant from a species of the genus *Flavorum* to the subject, wherein the plant extract is an extract of a species of *Myrothamnus flabellifolia*, and wherein the extract is obtained by solid / liquid extraction using subcritical water as the extraction solvent.

7. The method of claim 6, wherein the hair is eyelash or eyebrow hair.

8. The method of claim 7, wherein the method is a non-therapeutic cosmetic method for lengthening the hair of the eyelashes.

9. The method of claim 7, wherein the method is a non-therapeutic cosmetic method for increasing the density of the hairs in the eyebrows.

10. The method of claim 6, wherein the method is a non-therapeutic cosmetic method for increasing hair adhesion.

11. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the extract is obtained from the aerial parts of the Myrothamnus species.

12. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the plant extract comprises: mirtinoside and kaempferol-3-O-glucuronide; or mirtinoside, kaempferol-3-O-glucuronide and trehalose; or mirtinoside, kaempferol-3-O-glucuronide, trehalose and luteolin-7-O-glucuronide; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide and arbutin.

13. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the plant extract comprises mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtrin, kaempferol-3-O-glucuronide, seaweed Sugar, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, and sprucein; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, sprucein, and coniferaldehyde; or mirtinoside, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, sprucein, coniferaldehyde, and naringenin-7-O-glucoside.

14. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the plant extract comprises at least one compound selected from the group consisting of isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside, coniferaldehyde, and naringenin-7-O-glucoside.

15. The use according to claim 14, or the method according to claim 14, wherein the plant extract comprises isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside, coniferaldehyde, and naringenin-7-O-glucoside.

16. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the plant extract is applied topically by contact with hairy skin.

17. The use according to claim 5, or the method according to claim 7, wherein the plant extract is applied by applying it to the eyelash line and / or the eyebrows.

18. The use according to claim 17, or the method according to claim 17, wherein the plant extract is applied by applying it to the eyelashes.

19. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein hair growth and / or increased hair thickness and / or prevented hair loss are promoted by: inducing proliferation of dermal papilla cells in the hair follicle; inducing proliferation of keratinocytes and / or associated keratin to form hair; increasing blood flow in the hair follicle; and / or increasing the synthesis of XVII collagen and α6β4 integrin in the hair follicle.

20. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the extract is in a composition further comprising a water-miscible organic solvent.

21. The use according to claim 20, or the method according to claim 20, wherein the organic-water miscible solvent is a polyol.

22. The use according to claim 21, or the method according to claim 21, wherein the polyol is a C2-C10 aliphatic hydrocarbon diol or glycerol.

23. The use according to claim 20, or the method according to claim 20, wherein the weight ratio of the extract to the organic miscible solvent is 1:1 to 1:

10.

24. The use according to any one of claims 1 to 4, or the method according to any one of claims 6 to 10, wherein the extract is in a cosmetic composition comprising at least one cosmetically acceptable ingredient.

25. The use according to claim 24, or the method according to claim 24, wherein the cosmetic composition is selected from the group consisting of lotion, shampoo, serum, or mascara compositions.

26. The use according to claim 25, or the method according to claim 25, wherein the composition is in the form of a serum for eyebrows or eyelashes, or in the form of a mascara composition for eyelashes.

27. The use according to claim 26 or the method according to claim 26, wherein the concentration of the extract is from 0.5% (w / w) to 5% (w / w).

28. The use according to claim 24, or the method according to claim 24, wherein the at least one cosmetically acceptable ingredient is selected from the group consisting of: vitamins, minerals, proteins, peptides, fatty acids, antioxidants, anti-inflammatory agents, color enhancers and / or mixtures thereof.

29. A method for obtaining a plant extract from a Myrothamnus species for use in any one of claims 1 to 4 or the method in any one of claims 6 to 10, the method comprising extracting the plant material of the Myrothamnus species with subcritical water, wherein the extraction comprises the following steps: i) To form a plant extract, the plant material from the Myrothamnus species is brought into contact with the water for at least 10 minutes at a temperature of 120°C to 220°C and at a pressure suitable for maintaining the subcritical water in a liquid state. as well as ii) Separate the plant material from the plant extract.

30. The method of claim 29, wherein the plant material comprises the stems and / or leaves of the plant.

31. The method of claim 29, wherein the plant material is dried prior to extraction.

32. The method according to claim 31, wherein the weight ratio of the dried plant material to the volume of water is from 1:10 g / mL to 1:40 g / mL.

33. The method of claim 29, wherein the temperature is 140°C to 160°C.

34. The method of claim 29, further comprising the step of mixing the plant extract with a water-miscible organic solvent.

35. The method according to claim 34, wherein the water-miscible organic solvent is a polyol.

36. The method according to claim 35, wherein the polyol is a C2-C10 aliphatic hydrocarbon diol or glycerol.

37. The method according to claim 34, wherein the concentration ratio of plant extract to water-miscible organic solvent by weight is 1:1 to 1:

10.

38. A plant extract for use in any one of claims 1 to 4 or in any one of claims 6 to 10, said plant extract being obtainable by any one of claims 29 to 33.

39. The plant extract according to claim 38, wherein the plant extract comprises at least one compound selected from the group consisting of isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside, coniferaldehyde, and naringenin-7-O-glucoside.

40. The plant extract according to claim 39, wherein the plant extract comprises isorhamnetin-3-O-glucoside, naringenin, spruce glycoside, coniferaldehyde, and naringenin-7-O-glucoside.

41. The plant extract according to claim 40, wherein the concentrations of isorhamnetin-3-O-glucoside, naringenin, spruce neoglycoside, coniferaldehyde and naringenin-7-O-glucoside are each at least 0.05 ppm.

42. A composition for use in any one of claims 1 to 4 or in any one of claims 6 to 10, said composition comprising the plant extract of claim 38 and a water-miscible organic solvent.

43. The composition of claim 42, further comprising additional water.

44. The composition according to claim 42, wherein the water-miscible organic solvent is a polyol.

45. The composition according to claim 44, wherein the polyol is a C2-C10 aliphatic hydrocarbon diol or glycerol.

46. ​​The composition of claim 44, wherein the polyol is present in an amount of 50% to 90% by weight based on the total composition.

47. A cosmetic composition for use in any one of claims 1 to 4 or the method of any one of claims 6 to 10, the cosmetic composition comprising: the plant extract of claim 38, or the composition of claim 42; and at least one cosmetically acceptable ingredient.

48. The cosmetic composition of claim 47, wherein the cosmetic composition comprises the composition of claim 42, and the amount of the composition is from 0.01% by weight to 20% by weight.

49. The cosmetic composition according to claim 48, wherein the amount of the composition according to claim 42 is from 0.5% by weight to 4% by weight.

50. The cosmetic composition according to claim 48, wherein the amount of the composition according to claim 42 is from 1% to 3% by weight.

51. The cosmetic composition of claim 47, wherein the at least one cosmetically acceptable ingredient is selected from surfactants, hair conditioners, emollients, emulsifiers, humectants, rheology modifiers, vitamins, hair growth promoters, anti-dandruff agents, moisturizers, hair fixatives, hair colorants, oxidants, hair dyes, hair bleaching agents, pigments; and combinations thereof.

52. The cosmetic composition according to claim 47, wherein the cosmetic composition is in the form of a product selected from the group consisting of: creams, complex emulsions, solutions, liquid crystals, anhydrous compositions, aqueous dispersions, oils, balms, foams, and gels.

53. The cosmetic composition according to claim 47, wherein the cosmetic composition is in the form of a product selected from the group consisting of: creams, gels, hydroalcoholic solutions, hydroglycol solutions, hydrogels, lotions, lotions, soaps, shampoos, conditioners, serums, polysaccharide films, ointments, mousses, hair oils, powders, sticks, pens, sprays, or aerosols.

54. A mascara composition for use in any one of claims 1 to 4 or the method of any one of claims 6 to 10, the mascara composition comprising a plant extract of the species Myrothamnus flabellifolia and at least one wax, wherein the extract is obtained by solid / liquid extraction using subcritical water as the extraction solvent.

55. The mascara composition of claim 54, further comprising a liquid phase and at least one emulsifier.

56. The mascara composition according to claim 54 or claim 55 further comprises at least one rheology modifier.

57. The mascara composition of claim 56, wherein the at least one rheology modifier comprises a thickener and / or a film-forming agent.

58. The mascara composition of claim 54, wherein the at least one wax is selected from the group consisting of: rice wax, rice bran wax, carnauba wax and shellac wax.

59. The mascara composition according to claim 54, wherein the mascara composition comprises at least one pigment.

60. The mascara composition according to claim 59, wherein the pigment is a metal oxide.

61. The mascara composition of claim 55, wherein the mascara composition comprises at least one emulsifier selected from the group consisting of fatty acids, fatty acid esters of glycerol and / or polyalkylene glycols, amphoteric acetates and alkyl phosphate esters.

62. The mascara composition of claim 57, wherein the mascara composition comprises at least one thickener selected from the group consisting of: a crosslinked homopolymer of acrylic acid or methacrylic acid, and microbial polysaccharides.

63. The mascara composition according to claim 62, wherein the microbial polysaccharide is a dermal filler.

64. The mascara composition of claim 57, wherein the mascara composition comprises at least one film-forming agent selected from the group consisting of: acrylic copolymers, polyethylene polymers, and copolymers of polyvinylpyrrolidone (PVP), ethylene-vinyl acetate, polydimethylsiloxane, and polyurethane.

65. The mascara composition of claim 54, wherein the plant extract is obtained from the aerial parts of the Myrothamnus flabellifolia species.

66. The mascara composition of claim 54, wherein the plant extract comprises: mirtrin and kaempferol-3-O-glucuronide; or mirtrin, kaempferol-3-O-glucuronide and trehalose; or mirtrin, kaempferol-3-O-glucuronide, trehalose and luteolin-7-O-glucuronide; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide and arbutin.

67. The mascara composition according to claim 54, wherein the plant extract comprises mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mirtrin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucoside, and naringenin. Glucuronyl glycoside, isorhamnetin-3-O-glucoside, naringenin and sprucein; or mirtinoside, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, isorhamnetin-3-O-glucoside, naringenin, sprucein and coniferaldehyde; or mirtinoside, kaempferol-3-O-glucoside, trehalose, luteolin-7-O-glucoside, isorhamnetin-3-O-glucoside, naringenin, sprucein, coniferaldehyde and naringenin-7-O-glucoside.

68. The mascara composition of claim 54, wherein the plant extract comprises at least one compound selected from the group consisting of isorhamnetin-3-O-glucoside, naringenin, spruce glycoside, coniferaldehyde, and naringenin-7-O-glucoside.

69. The mascara composition of claim 68, wherein the plant extract comprises isorhamnetin-3-O-glucoside, naringenin, spruce glycoside, coniferaldehyde, and naringenin-7-O-glucoside.

70. The mascara composition of claim 54, wherein the plant extract is the plant extract of claim 38.

71. The mascara composition of claim 70, wherein the plant extract is in a composition further comprising a water-miscible organic solvent.

72. The mascara composition according to claim 71, wherein the organic water-miscible solvent is a polyol.

73. The mascara composition according to claim 72, wherein the polyol is a C2-C10 aliphatic hydrocarbon diol or glycerol.

74. The mascara composition according to claim 71, wherein the weight ratio of the plant extract to the organic miscible solvent is 1:1 to 1:

10.

75. The mascara composition of claim 71, wherein the plant extract is in the composition of claim 42.

Images