Use of Myrothamnus species extract to promote hair growth

Abstract

Use of a plant extract of Myrothamnus species for promoting and / or increasing hair growth and / or preventing hair loss and / or increasing hair thickness, particularly of eyelashes and / or eyebrows. Methods for preparing the plant extract and compositions comprising same are also disclosed.

Description

[Technical field]

[0001] The present invention relates to the use of a plant extract of Myrothamnus species to promote hair growth and / or prevent hair loss and / or increase hair thickness. In one embodiment, the hair is eyebrow and / or eyelash hair. Methods for preparing the plant extract and compositions comprising same are also disclosed. [Background technology]

[0002] Hair, particularly eyelashes and eyebrow hair, plays an important role in human aesthetics. Eyelashes and eyebrows are prominent features of the face. They serve to protect the eyes and influence facial expression. These hairs grow relatively slowly and have a short average length. Many cultures place a special emphasis on and desire for long and thick eyelashes. There is an interest in improving the appearance of eyelashes by increasing the length and thickness of eyelashes. Additionally, there is an interest in improving the appearance of a person's eyebrows by increasing the thickness of eyebrow hairs. Hair damage and / or loss is a common problem in human subjects and can have a significant impact on the overall appearance of a person. Eyelash and eyebrow hairs are particularly susceptible to damage due to finger / hand contact, for example when a subject rubs their eyes with their hands when tired. Mechanical or chemical stress during styling can weaken and damage the hair. Hair loss can have many other causes (e.g., genetic predisposition, aging, and / or disease). There is great interest in treatments that can increase hair strength, prevent hair loss, or promote hair growth.

[0003] Hair grows from dermal follicles that extend from the epidermis (the outermost layer of the skin) to the dermis (the deepest layer of the skin). The length and thickness of hair depends on two main biological processes: hair follicle renewal and hair fiber synthesis.

[0004] The hair cycle is a stem cell-mediated process that occurs in adult skin and involves the cyclic destruction and regeneration of hair follicles. The hair cycle consists of three defined phases: growth (anagen), followed by regression (catagen), and rest (telogen). Anagen is the longest phase of the hair cycle. During this phase, cell proliferation begins, hair follicles are formed, and hair is continuously produced. During catagen, cessation of proliferation and increased cell apoptosis occurs. Hair stops growing and the hair follicle shortens. Finally, during the telogen phase, also described as the resting phase, the hair is shed. [Alonso L and Fuchs E (2006) The hair cycle. J. Cell Sci. 119:391-393].

[0005] The growth of new hair requires re-entry into anagen, a process that involves the activation of multipotent epithelial stem cells that reside in a specialized portion of the hair follicle outer root sheath (ORS) known as the bulge. Dermal cells known as hair follicle dermal papilla cells (HFDPCs) reside in the bulge [Beaudoin GM 3rd, Sisk JM, Coulombe PA and Thompson CC. (2005) Hairless triggers reactivation of hair growth by promoting Wnt signaling. Proc Natl Acad Sci USA. 102-14653-14658]. HFDPCs in adult hair follicles play a key role in the hair growth cycle by inducing hair follicle development [Drikell RR, Clavel C, Rendl M and Watt FM. (2011) Hair follicle dermal papilla cells at a glance. J. Cell Sci. 124:1179-1182].

[0006] Hair fibers are composed primarily of proteins (65-95% of hair weight), with keratin being the most abundant. The fiber consists of three general layers: the cortex, the hair cuticle, and the medulla. The central medulla contains polygonal cells with a sponge-like appearance. Around the medulla, the cortex forms a layer of keratinized fiber cells with a longitudinal orientation and are filled with keratin filaments. In addition, the cortex contains melanosomes, which determine the color of the hair fiber. The hair cuticle, the outermost layer of the hair fiber, consists of multiple layers of keratinocytes. The hair cuticle is thin and translucent, allowing light to penetrate to the cortical pigments. The keratin proteins that form this complex structure of the hair fiber are the result of the proliferative activity of keratinocytes located in the hair follicle. These keratinocytes are found in the dermal papilla (DP) and develop hair growth along with other cells involved in this proliferative activity.

[0007] Compositions and methods for promoting hair growth are known in the art, however, known methods may be ineffective or may have undesirable side effects.

[0008] Drugs including Minoxidil® (Rogaine), Finasteride® (Propecia), and Dutasteride® (Avodart) are approved treatments for hair loss. However, their effectiveness is mainly limited to hormone-related alopecia (i.e., androgenic alopecia), and they require a medical prescription. Also, there is growing concern about the use of synthetic chemicals in cosmetic preparations applied to the human body. Because the human eyeball and surrounding membranes are particularly sensitive compared to human skin on other areas of the body, developing cosmetic preparations for areas in close proximity to the human eye, such as preparations designed to promote eyelash or eyebrow growth, presents particular challenges. Although cosmetics for eyelashes are not intended to be applied directly to the eyeball or eye membranes, users may inadvertently transfer some of the cosmetic preparation into the eyeball during application. Additionally, there is growing concern about the environmental impacts caused by the production of synthetic chemical products utilized in cosmetics, for example, by washing cosmetics off the body after use, or by disposing of unused amounts of such products in landfills.

[0009] Mascara, which adds volume to eyelashes, is one of the most common products for enhancing the appearance of eyelashes. However, those with short eyelashes may find it difficult to achieve thicker looking eyelashes with mascara alone and may choose to use eyelash extensions. Eyelash extensions often involve the use of rough adhesives to bond the extensions to the eyelids near the lash line, and thus may cause damage to natural eyelashes. Various risks associated with the use of eyelash extensions have been reported, such as eyelid swelling, loss of eyelashes, eyelid skin irritation, or eyelid infection. A mascara composition that can promote eyelash growth and improve the appearance of eyelashes is desired.

[0010] Microtinting and microblading are tattooing procedures used to improve eyebrow density and definition. However, they cause the inconveniences associated with any tattooing procedure, such as pain and risk of infection.

[0011] There is a need for natural, sustainable and effective solutions to the above problems. Several herbal preparations have been proposed to promote hair growth in eyelashes and eyebrows. However, their effectiveness may be limited.

[0012] European Patent No. 2764894B1 provides an extract from the leaves of the blackberry plant for inducing hair growth. The extract is said to induce a visible telogen phase (i.e., shedding of club hairs) before a rapid entry into anagen phase (i.e., the active growth phase of the hair follicle). Extractants used to obtain the extract include ethanol or a combination of ethanol and alcohol. Ethanol and alcohol extraction have inherent flammability hazards in manufacturing and production as well as transportation logistics. Ethanol is considered an environmentally friendly solvent and has good extraction capabilities, but is undesirable for some segments of the cosmetic market and for some consumers.

[0013] US Patent Application Publication No. 20190224160A1 discloses a composition for promoting eyelash growth comprising at least forskolin derived exclusively from Coleus forskohlii in combination with a keratinocyte growth stimulator comprising at least one botanical extract selected from the group comprising Tussilago farfara flower extract, Achillea millefolium extract, Cinchona succirbra bark extract, Nasturtium officinale, and Tropaeolum majus.

[0014] WO2017032711 discloses a cosmetic composition for promoting eyelash growth, comprising a plant extract of mung bean (Vigna radiata), an extract of mung bean sprouts, and a peptide comprising, from the N-terminus to the C-terminus, the amino acid sequence Gly-His-Lys.

[0015] Myrothamnus species are drought tolerant shrubs native to South Africa that belong to the family of plants known as "foot-and-foot lilies". The plant is adapted to the seasonal wet and dry periods of the region. In the dry periods, the leaves dry out as the plant goes dormant while it waits for the rains to return. At the first rainfall, the plant revives and bursts into life, rapidly hydrating the leaves and flowering. Myrothamnus has many ways of avoiding mechanical and drought stress and is of interest to the scientific community looking for active substances with cosmetic and / or medicinal properties. Myrothamnus is consumed as a tea in South Africa and the essential oil is also extracted for use. Its use in local food and medicinal traditions is well known in South Africa. Myrothamnus is used in the cosmetics industry for skin care and is the subject of several patents / patent applications such as US200701341934A1, FR29978536B1 and KR1305698B1. There is a need to find natural cosmetic non-therapeutic treatments that can increase hair growth and thickness, especially for hair in sensitive facial areas such as near the eyes, i.e., eyelashes and / or eyebrows. Desirably, such treatments have little or no negative side effects and / or provide cosmetic benefits to the hair and its associated skin. The present invention aims to meet some or all of these needs and to solve some or all of the problems identified above. [Prior art documents] [Patent documents]

[0016] [Patent Document 1] European Patent No. 2764894 [Patent Document 2] US Patent Application Publication No. 2019 / 0224160 [Patent Document 3] International Publication No. 2017 / 032711 [Patent Document 4] US Patent Application Publication No. 2007 / 01341934

Patent Document 5

Patent Document 6

Non-Patent Document

[0017]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Summary of the Invention

Means for Solving the Problems

[0018] In a first aspect, the present invention provides the use of an extract of Myrothamnus sp. for promoting hair growth, preventing hair loss, and / or increasing hair thickness. In particular, the use may be a cosmetic non-therapeutic use. In particular, the hair may be eyelash and / or eyebrow hair. The present invention provides the cosmetic non-therapeutic use of an extract of Myrothamnus sp. for increasing the length of eyelashes and / or for increasing eyebrow thickness (i.e., for increasing hair density). The use of Myrothamnus extract is particularly advantageous, since cosmetic treatments considered "natural" are generally environmentally friendly, and consumers have a particular interest in such treatments.

[0019] In another aspect, the present invention provides a method for promoting and / or increasing hair growth, preventing hair loss, and / or increasing hair thickness, comprising administering to a subject a plant extract of Myrothamnus sp. In particular, the method can be a cosmetic non-therapeutic method. In particular, the hair can be eyelash or eyebrow hair. The present invention provides a cosmetic non-therapeutic method for increasing eyelash length and / or increasing eyebrow thickness (i.e., increasing hair density) comprising administering to a subject a plant extract of Myrothamnus sp.

[0020] In another aspect, the present invention provides a process for obtaining a plant extract from Myrothamnus sp., comprising subjecting a plant material of Myrothamnus sp. to extraction with subcritical water, the extraction comprising: i) contacting the plant material with subcritical water at a temperature of at least 120° C. and at a pressure sufficient to maintain the water in a liquid state for at least 10 minutes to form an aqueous plant extract; ii) separating the plant material from the aqueous plant extract.

[0021] In another aspect, the present invention provides an aqueous plant extract obtained / obtainable by the above-mentioned process for obtaining a plant extract from Myrothamnus species.

[0022] In another aspect, the present invention provides a cosmetic composition comprising an aqueous plant extract and at least one cosmetically acceptable excipient or ingredient.

[0023] In another aspect, the present invention provides a mascara composition comprising a plant extract from a Myrothamnus species and at least one pigment. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] The present invention is based on the discovery of surprising properties of plant extracts from plants of the Myrothamnus genus, i.e. Myrothamnus species, which properties render the extracts suitable for cosmetic, non-therapeutic applications.

[0025] definition In the context of the present invention, "hair" includes hair of the scalp, skin, eyelashes, eyebrows, moustache region, and / or beard region of a subject. In particular, hair is eyebrow and / or eyelash hair. By "skin" is understood the layers that make up the skin, including both the top layer or stratum corneum to the bottom layer or subcutaneous tissue. These layers are composed of different types of cells, such as keratinocytes, fibroblasts, melanocytes, mast cells, neurons, and / or adipocytes, among others. The term "skin" includes mammalian skin, e.g. human skin, including skin containing hair.

[0026] As used herein, the term "about," for example when referring to a measurable value (e.g., an amount or weight of a particular ingredient, or temperature), refers to a variation of ±20%, ±10%, ±5%, ±1%, ±0.5%, or especially ±0.1% of the specified amount.

[0027] As used herein, the term "comprising" is inclusive or open-ended and does not exclude additional, unrecited elements or method steps, and is intended to encompass the phrases "consisting essentially of" and "consisting of" as alternative embodiments, where "consisting of" excludes any elements or steps not specified, and "consisting essentially of" permits the inclusion of additional, unrecited elements or steps that do not materially affect the essential or basic and novel characteristics of the composition or method under consideration.

[0028] extract The present invention relates to the use of plant extracts of Myrothamnus species. The terms "plant extract" and "extract" are used interchangeably herein and refer to a product containing one or more compounds extracted from plant material of Myrothamnus species. Specifically, the product is obtained by subjecting plant material of Myrothamnus species to solid / liquid extraction, whereby one or more phytochemical compounds contained in the plant material (solid) are extracted from the plant material into a solvent (liquid) and they elute from the plant material into the solvent. The product may be a composition comprising an extraction solvent and one or more phytochemical compounds. The solid / liquid extraction process may include a separation step, optionally followed by one or more purification steps. Thus, the extract may be the product obtained after the separation step or the product obtained after the purification step. In the separation step, a composition comprising the solvent and one or more phytochemical compounds is separated from the plant material. The extract is a separated composition comprising the extraction solvent and one or more phytochemical compounds. The solubility of the phytochemical compounds in the solvent may be temperature dependent, and thus the composition may be in solution or some of the botanical compounds may precipitate when cooled to a temperature lower than that used for extraction. When the solvent is water, the composition comprising the extraction solvent and one or more phytochemical compounds is also referred to herein as an "aqueous plant extract" or "aqueous extract." Purification (e.g., by fractionation, concentration, or drying) of the composition comprising the solvent and one or more phytochemical compounds provides an extract, which may be, for example, a concentrate (e.g., a viscous form) or a solid (e.g., a powder).

[0029] Myrothamnus species is a genus of small xerophytic shrub flowering plants that includes two species, namely M. flabellifolia and M. moschata. In particular, the extract may be an extract of M. flabellifolia, which is also referred to in the literature as M. flabellifoliaus.

[0030] Typically, the plant material from Myrothamnus species comprises the above-ground parts of the plant, in particular the stems and / or leaves. The plant material may comprise the stems and / or leaves as the main component, for example the stems and / or leaves constitute more than 50, 80 or 90% by weight of the plant material. The plant material may comprise only the stems and / or leaves, i.e. they are the only components and no other plant material is present.

[0031] Typically, the plant material is in a dry form. The plant material can be dried to make it easier to homogenize, manipulate, and store. By dried plant material or plant material in a dry form is meant plant material that has been dried to have a moisture content of less than 10% water by weight, less than 5% water by weight, less than 2% water by weight, or less than 1% water by weight. The moisture content can be measured, for example, by the AOAC (Association of Official Analytical Chemists) official method AOAC 934.06-1934 (1996). However, non-dried plant material may also be used.

[0032] Advantageously, the plant material is ground or crushed before extraction. The ground plant material can have an average particle size of 0.01 or 0.1 to 10 mm, for example with a particle size in the range of 100 pm to 50 mm. More specifically, the average particle size is in the range of 0.2 to 5 mm. The average particle size can be determined by conventional methods, for example methods involving sieve analysis. Any suitable grinding / crushing technique known in the art can be used to obtain the desired particle size of the plant material. Particularly suitable plant material is dry plant material in powder form, for example with a particle size as mentioned above.

[0033] The plant material can be mixed with a neutral material to make it drier and / or more porous for solvent extraction. Suitable neutral materials include graphene, silica gel, C18 resin, diatomaceous earth, and neutral alumina.

[0034] Suitable solvents for use in extraction include water, lower alcohols of 1 to 4 carbon atoms (e.g., methanol, ethanol, butanol, etc.) and glycols, and combinations thereof. Advantageously, an extract from Myrothamnus species can be obtained using water as the only extraction solvent. In this case, compounds in the plant material are dissolved from the plant material in water to form an aqueous extract (also referred to herein as an aqueous plant extract). The aqueous extract can be purified to form an extract that is, for example, a concentrate or a powder. Thus, advantageously, an extract from Myrothamnus species can be obtained avoiding the use of organic solvents during the extraction process.

[0035] Any conventional solid / liquid extraction method, such as Soxhlet, percolation, and immersion, can be used to obtain a plant extract from the plant material of Myrothamnus species. The temperature of the solvent should be selected to be appropriate for that solvent. When water is used as the extraction solvent, it is preferably used at a temperature above 60°C. Typically, the ratio of dry plant material / solvent (weight (g) of dry plant material / volume (mL) of solvent) ranges from 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, the ratio is 1:10 or 1:20.

[0036] In one embodiment, the extract of Myrothamnus species is obtained by subjecting the plant material of Myrothamnus species to a solid / liquid extraction using subcritical water as the extraction solvent. Subcritical water is water that is held in a liquid state by pressure at a temperature above its natural boiling point of 100°C (i.e., above its boiling point at atmospheric pressure). Subcritical water can have a temperature up to a critical point temperature of 374°C. Subcritical water is also called "pressurized low polarity water", "compressed hot water" or "compressed hot water". When water is heated under pressure to a temperature above its boiling point, its important properties, such as polarity, are changed. Preferably, when subcritical water is used as the extraction solvent, it is the only extraction solvent used in the process, i.e., the extraction is carried out with subcritical water in the absence of any other solvent (organic or inorganic).

[0037] More specifically, the extract from Myrothamnus spp. is obtained by subjecting plant material of Myrothamnus spp. to extraction with subcritical water, the extraction comprising contacting the plant material with subcritical water at a temperature of at least 120° C. and at a pressure suitable to maintain the water in a liquid state for at least 10 minutes to form an aqueous plant extract. i) contacting plant material of Myrothamnus species with subcritical water at a temperature of at least 120° C. and at a temperature and pressure sufficient to maintain the water in a liquid state for at least 10 minutes to form an aqueous plant extract; ii) separating the plant material from the aqueous plant extract.

[0038] Typically, the ratio of dry plant material / subcritical water (weight of dry 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, the ratio may be 1:10 or 1:20.

[0039] In particular, the extraction can be carried out using subcritical water at a temperature of about 120 to about 220° C., about 130 to about 180° C., about 140 to about 160° C., or about 145 to about 155° C. In particular, the subcritical water can be at a temperature of about 150° C.

[0040] When subcritical water is the extraction solvent, the extraction is carried out in a pressurizable vessel, typically a stainless steel vessel. During the extraction, the pressure in the pressurizable vessel must be such that the subcritical water is maintained in a liquid state. The pressure required to achieve this varies depending on the temperature of the water. A person skilled in the art will be able to determine the necessary pressure. Typically, the pressure ranges from 0.5 MPa to 20 MPa. The pressure may be at least about 1 MPa. The pressure may be about 5 to about 15 MPa or about 8 to about 13 MPa. The pressure may be 10, 11, and 12 MPa.

[0041] The step of contacting the plant material with subcritical water is carried out for at least about 10 minutes. This time is the time during which the plant material is in contact with the subcritical water, and the water may be stationary or flowing over the plant. This period is also referred to herein as the extraction time. The extraction time varies depending, for example, on the amount of plant material and water used. Typically, the extraction time varies 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, more specifically from 20 minutes to 90 minutes. The extraction time may be from about 45 to 75 minutes or about 1 hour.

[0042] Subcritical water extraction can be performed in batch mode (also called "static mode") or dynamic mode (also called "flow-through"). In batch mode, the plant material is exposed to (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 plant material samples. In batch mode, the extraction time is the total time the plant material is exposed to subcritical water over all batches of plant material samples. 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 samples. In dynamic mode, the extraction time is the total time the plant material is exposed to the flow of subcritical water for the plant material samples. For faster and larger scale production of plant extracts, the extraction is advantageously performed in dynamic mode. The extraction can be performed in any system known in the art that allows for subcritical water extraction, examples include systems including batch extractors or continuous extractors.

[0043] During the step of contacting the plant material with subcritical water, components are extracted from the plant material into the subcritical water. More specifically, the plant material dissolves in the subcritical water, thereby forming an aqueous plant extract. Similarly, conventional solvent extraction methods using water as a solvent form an aqueous plant extract. The solvent extraction process can include a step of separating the plant material (i.e., any undissolved plant material) from the plant extract. Typically, this step is simple. For example, in the case of subcritical water extraction, it can simply involve releasing the aqueous plant extract from a pressurizable vessel while retaining the plant material in the pressurizable vessel. Other suitable separation techniques are known in the art and include, for example, filtration, sedimentation, decantation, or centrifugation. Filtration can be performed using a filter having a pore size of 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 successive filtration operations, for example using multiple filters with decreasing pore size. The residue (undissolved plant material) remaining after filtration may be recontacted with more extraction solvent. This filtration-recontacting step may be performed once or may be repeated, for example, 1-5 times. In the case of subcritical water extraction, the separation is preferably performed before the aqueous plant extract cools to precipitate the extract components.

[0044] After the plant extract is separated from the plant material, it can be purified. In the context of the present invention, "purify" means purification, partial purification, and / or fractionation. There are many techniques well known in the art 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 phase 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 10% or less water by weight, 5% or less water by weight, 2% or less water by weight, or 1% or less water by weight. This can be measured, for example, by the moisture content determination method 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 lyophilization (i.e., freeze drying), spray drying, atomization, or fluidized bed dryer. Concentration or drying can be performed on the plant extract with or without a carrier or other excipient. The extract can be a concentrate, or a solid, for example, an amorphous solid, a crystalline or partially crystalline solid, and can optionally be in the form of a powder.

[0045] Thus, the present invention also provides a Myrothamnus species plant extract obtained / obtainable by any of the above processes. In particular, the Myrothamnus species plant extract is obtained / obtainable by any of the above processes including extraction with water. The Myrothamnus species plant extract may be obtained / obtainable by any of the above processes including extraction with subcritical water. The details of the extraction process described herein may be applied or incorporated into the definition of the plant extract provided herein.

[0046] Plant extracts contain phytochemicals, which are compounds produced by plants, and can include, for example, polyphenols, amino acids, organic acids, and sugars.

[0047] The plant extract may be selected from the group consisting of mychellianin and kaempferol-3-O-glucuronide; mychellianin, kaempferol-3-O-glucuronide, and trehalose; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and arbutin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and arbutin. glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and kaempferol-3-O-glucoside; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, and quercetin-3-O-galactoside; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, Arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, and syringic acid; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, and quercitrin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercitrin, isoquercitrin and tryptamine.

[0048] The plant extract may be / can be obtained by solid / liquid extraction using water as a solvent and may be: mychellianin and kaempferol-3-O-glucuronide; mychellianin, kaempferol-3-O-glucuronide, and trehalose; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide. nide, and arbutin; or michellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and kaempferol-3-O-glucoside; or michellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, and quercetin-3-O-galacto .... tetrose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, and syringic acid; or michellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, and quercitrin; or michellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercitrin, isoquercitrin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, kaempferol-3-O-glucoside, quercetin-3-O-galactoside, syringic acid, quercitrin, isoquercitrin and tryptamine.

[0049] The plant extract may be selected from the group consisting of mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin. lingenin, and piceid; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, piceid, and coniferaldehyde; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

[0050] The plant extract may be / can be obtained by solid / liquid extraction using water as a solvent and may be selected from the group consisting of mycelianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mycelianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mycelianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, piceid, and coniferaldehyde; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

[0051] The plant extract may be selected from the group consisting of mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and isorhamnetin-3-O-glucoside; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, and naringenin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, and naringenin. sid, naringenin, and piceid; or mychellianin, kaempferol-3-O-glucuronide, trehalose and luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, naringenin, piceid, and coniferaldehyde; mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

[0052] The plant extract may be / is obtainable by solid / liquid extraction using water as a solvent and may be selected from the group consisting of mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, and isorhamnetin-3-O-glucoside; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, and naringenin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide, Arbutin, isorhamnetin-3-O-glucoside, naringenin, and piceid; or mychellianin, kaempferol-3-O-glucuronide, trehalose and luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, naringenin, piceid, and coniferaldehyde; mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, arbutin, isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

[0053] composition As mentioned above, the plant extract obtained from Myrothamnus species by extraction with water may be in aqueous form, i.e. in the form of an aqueous plant extract. The components extracted from the plant material may or may not be completely dissolved in the aqueous plant extract. Advantageously, the aqueous plant extract may be mixed with a water-miscible organic solvent to keep the extracted components in solution. The process for obtaining the plant extract of the present invention may comprise a step comprising mixing the 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 this composition to obtain the required concentration of the water-miscible solvent. The process for obtaining the plant extract of Myrothamnus species described herein may comprise a step comprising mixing the plant extract as defined herein with a water-miscible organic solvent and optionally water. The resulting composition comprises the plant extract together with the water-miscible organic solvent and optionally water. These solutions may be used as "stock solutions" and may be used, for example, for the preparation of cosmetic compositions.

[0054] The water-miscible organic solvent is preferably a cosmetically acceptable organic solvent and may be a polyol. Suitable polyols include glycols, which are organic compounds containing two alcohol functional groups (-OH groups), such as C2-C10 aliphatic hydrocarbyl diols or triols, and glycerin. C2-C10 aliphatic hydrocarbyl diols include C2-C10 or C2-C8 alkanediols in their respective isomeric forms, which may be substituted or unsubstituted. If substituted, the alkanediol may be substituted with one or more substituents independently selected from, for example, halo groups, hydroxyl groups, ester groups, 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, butylene glycol, pentylene glycol, glycerol, or caprylyl glycol, and mixtures thereof. In particular, the polyol is glycerol (also referred to herein as glycerine or glycerin).

[0055] A composition comprising a plant extract and a polyol and optionally (additionally) water can 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 remainder is made up of the plant extract and water. For example, the composition can contain up to 50% by weight of water. Preferably, the polyol is glycerin. A composition comprising a plant extract and a polyol and water can 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 remainder is made up of the plant extract and water. For example, the composition can contain up to 50% by weight of water. Preferably, the polyol is glycerin.

[0056] The extract, or a composition comprising the extract, a water-miscible organic solvent, and optionally water, as described above, can be incorporated into a composition suitable for administration to a subject.

[0057] The composition may be a cosmetic composition comprising the extract together with at least one cosmetically acceptable excipient or adjuvant, or a composition comprising the extract, a water-miscible organic solvent, and optionally water, as described above. These compositions may be prepared by conventional means known to those skilled in the art ["Harry's Cosmeticology", Seventh edition, (1982), Wilkinson JB, Moore RJ, ed. Longman House, Essex, GB].

[0058] The cosmetic composition contains a cosmetically effective amount of the plant extract to be administered, as well as the dosage of the plant extract, which depends on a number of factors, including the age, condition of the patient, the nature or severity of the condition, the disorder or disease being treated and / or cared for, the route and frequency of administration, and the particular nature of the compound used.

[0059] The term "cosmetically effective amount" is understood to mean a non-toxic but sufficient amount of the extract of the present invention to provide the desired effect. The extract or stock solution of the present invention is used in the cosmetic composition of the present invention at a cosmetically effective concentration to achieve the desired effect, for example, in an amount of 0.00000001% (wt) to 20% (wt), 0.000001% (wt) to 15% (wt), 0.00001% (wt) to 10% (wt), or 0.0001% (wt) to 5% (wt), or 0.1 to 4% (wt), or 1 to 3% (wt) based on the total weight of the composition.

[0060] In one embodiment, the extract or the 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).

[0061] The cosmetic composition of the present invention may be a composition for topical application, optionally including cosmetically or pharma- ceutically acceptable excipients necessary for formulation into the desired dosage form. A topical composition is a composition suitable for topical application to mammalian keratinous tissue, such as hair-bearing skin, in particular the human scalp. In particular, topical compositions are hair care compositions, such as conditioners, treatments, hair tonics, styling gels, mousses, shampoos, hairsprays, pomades, setting lotions, coloring, and permanent wave compositions. Of particular relevance for the purposes of the present invention are tonics, conditioners, treatments, and styling gels, which may be in the form of gels, lotions, tinctures, sprays, mousses, cleansing compositions, or foams, and may be applied, for example, once a day as a lotion, tincture, mousse, or spray, or once or twice a week as a conditioner or treatment, depending on the individual needs. In particular, compositions suitable in the context of the present invention are lotions, shampoos, serums, or eyelash mascaras. Preferably, the cosmetic composition is a serum or eyelash mascara. The term "serum" is well known to those skilled in the art and refers to a composition that is transparent, gel-based, or liquid. Serums have a fluid texture and are more concentrated with active agents than standard care products. As used herein, the term "mascara" refers to a cosmetic product that is used to strengthen eyelashes. Mascara can, for example, darken, thicken, lengthen, and / or define eyelashes.

[0062] The cosmetic composition may be a mascara composition. The mascara composition of the present invention is similar to currently known mascaras in that it incorporates the basic formulation elements of a mascara.

[0063] 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. 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 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 further comprise at least one rheology modifier, such as a thickener and / or a film former.

[0064] As used herein, the term "wax" is intended to mean a lipophilic aliphatic compound that is solid at room temperature (about 25°C) and atmospheric pressure (760 mmHg, i.e. 105 Pa), undergoes a reversible solid / liquid state change, and has a melting point above 30°C, in some embodiments above about 55°C, up to about 120°C, or even about 200°C. The term wax includes waxes of animal origin, vegetable origin, mineral origin, and synthetic origin. Examples of waxes of animal origin include beeswax and lanolin wax. Examples of waxes of vegetable origin include rice wax, carnauba wax, candelilla wax, auricaria wax, cork fiber wax, sugarcane wax, Japan wax, sumac wax, cotton wax, sunflower wax. Examples of waxes of mineral origin include paraffin, microcrystalline wax, montan wax, ozokerite, and ceresin. Examples of waxes of synthetic origin include polyolefin waxes, such as polyethylene waxes, waxes obtained by Fischer-Tropsch synthesis, waxy copolymers and their esters, as well as silicone waxes and fluoro waxes. Suitable waxes include rice bran wax (Oryza Sativa Bran Cera), carnauba palm wax (Copernica Cerfifera) and shellac wax. The term wax may further include high melting point hydrogenated oils of animal or vegetable origin. Examples include C8-C 32 These include hydrogenated jojoba wax and hydrogenated oils obtained by catalytic hydrogenation of fats consisting of linear or non-linear fatty chains, hydrogenated sunflower oil, hydrogenated castor oil, hydrogenated copra oil, hydrogenated lanolin, and hydrogenated palm oil. The mascara composition may comprise at least one wax, and the total amount of wax in the composition is 0.5-20% by weight, or 1-10% by weight, or 2-8% by weight, or 4-7% by weight, based on the total weight of the mascara composition.

[0065] The pigments may be mineral and / or organic, coated or uncoated. Mineral pigments may include metal oxides, in particular titanium dioxide, zirconium oxide, zinc oxide or cerium oxide, optionally surface-treated, and iron oxides, in particular black iron oxide, titanium oxide or chromium oxide, manganese violet, ultramarine blue, chromium hydrate, and ferric blue. Organic pigments include carbon black, D&C type pigments, and lakes based on cochineal carmine or barium, strontium, calcium or aluminium. The mascara composition may comprise at least one pigment, the total amount of pigments in the composition being 1-20% by weight, or 5-15% by weight, or 8-12% by weight, based on the total weight of the mascara composition.

[0066] Suitable emulsifiers for use in mascara compositions include anionic, amphoteric, and nonionic emulsifiers suitable for emulsifying fatty compounds in the aqueous phase.In certain embodiments of the present invention, the emulsifier is selected from fatty acids, fatty acid esters of glycerol and / or polyalkylene glycols, amphoacetates, and alkyl phosphates.In certain embodiments of the present invention, the emulsifier is selected from stearic acid, glyceryl stearate, peg-200 glyceryl stearate, steareth-2, steareth-20 isoceteth-20, and potassium cetyl phosphate, and disodium cocoamphoacetate.Suitable emulsifiers include Glucate SS, Arlatone MAP160, and mixtures thereof. The mascara composition may comprise at least one emulsifier, and the total amount of emulsifier in the composition is from 0.01 to 8% by weight, or from 0.05 to 8% by weight, or from 0.05 to 5% by weight, or from 0.05 to 0.5% by weight, or from 0.1 to 0.20% by weight, based on the total weight of the mascara composition.

[0067] The mascara composition may include rheology modifiers, including viscosity-increasing polymeric natural and derivatized gums, resin thickeners, gelling agents, or suspending agents. To increase the viscosity, the composition may include one or more rheology modifiers, which may be synthetic or natural.

[0068] Examples include fatty alcohols such as C10-C32 alcohols, e.g., C12-C22 alcohols, natural oils, and polymers of acrylic and / or methacrylic acid, e.g., carbomers. Exemplary natural oils include mineral oils (mainly C15-C40 linear and branched aliphatic alkanes, with small amounts of cycloalkanes), naturally derived esters, natural alkanes, or vegetable oils. Exemplary synthetic rheology modifiers include acrylic polymers and copolymers. One class of acrylic rheology modifiers is the carboxyl-functional alkali-swellable and alkali-soluble thickeners (ASTs) produced by free radical polymerization of acrylic acid alone or in combination with other ethylenically unsaturated monomers. The polymers can be synthesized by solvent / precipitation and emulsion polymerization techniques. Exemplary synthetic rheology modifiers of this class include homopolymers of acrylic acid or methacrylic acid, and copolymers polymerized from one or more monomers of acrylic acid, substituted acrylic acid, and salts and C1-C30 alkyl esters of acrylic acid and substituted acrylic acid. As defined herein, the substituted acrylic acid contains a substituent located on the α and / or β carbon atom of the molecule, and in one aspect, the substituent is independently selected from C1-4 alkyl, -CN, and -COOH. Optionally, other ethylenically unsaturated monomers, such as, for example, styrene, vinyl acetate, ethylene, butadiene, acrylonitrile, and mixtures thereof, can be copolymerized into the backbone. The aforementioned polymers are optionally crosslinked by a monomer containing two or more moieties containing ethylenic unsaturation. In one aspect, the crosslinker is selected from polyalkenyl polyethers of polyhydric alcohols containing at least two alkenyl ether groups per molecule. Other exemplary crosslinkers are selected from allyl ethers of sucrose and allyl ethers of pentaerythritol, and mixtures thereof. These polymers are more fully disclosed in US Pat. Nos. 5,087,445, 4,509,949, and 2,798,053.

[0069] In one aspect, the AST rheology modifier or thickener is a crosslinked homopolymer polymerized from acrylic acid or methacrylic acid, commonly referred to by the INCI name of carbomer. Commercially available carbomers include Carbopol® polymers 934, 940, 941, 956, 980, and 996 available from Lubrizol Advanced Materials, Inc. In a further aspect, the rheology modifier is selected from a crosslinked copolymer polymerized from a first monomer selected from one or more monomers of acrylic acid, substituted acrylic acid, salts of acrylic acid, and salts of substituted acrylic acid, and a second monomer selected from one or more C10-C30 alkyl acrylate esters of acrylic acid or methacrylic acid. In one aspect, the monomers can be polymerized in the presence of a steric stabilizer as disclosed in U.S. Pat. No. 5,288,814, incorporated herein by reference. Some of the aforementioned polymers are designated under the INCI nomenclature as Acrylates / C10-30 Alkyl Acrylate Crosspolymers and are commercially available from Lubrizol Advanced Materials, Inc. under the trade names Carbopol® 1342 and 1382, Carbopol® Ultrez 20 and 21, Carbopol® ETD 2020, and Pemulen® TR-1 and TR-2.

[0070] In another embodiment, the rheology modifier can be a crosslinked linear poly(vinylamide / acrylic acid) copolymer as disclosed in U.S. Pat. No. 7,205,271, the disclosure of which is incorporated herein by reference.

[0071] Another class of synthetic rheology modifiers suitable for use in the compositions 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 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. The associative monomers include ethylenically unsaturated polymerizable end groups, a nonionic hydrophilic midsection that terminates in a hydrophobic end group. The nonionic hydrophilic midsection includes polyoxyalkylene groups, e.g., polyethylene oxide, polypropylene oxide, or a mixture of polyethylene oxide / polypropylene oxide segments. The terminal hydrophobic end groups are typically C8-C40 aliphatic moieties. Exemplary aliphatic moieties are selected from linear and branched alkyl substituents, linear and branched alkenyl substituents, carbocyclic substituents, aryl substituents, aralkyl substituents, arylalkyl substituents, and alkylaryl substituents. In one aspect, the associative monomers can be prepared by condensation of polyethoxylated and / or polypropoxylated aliphatic alcohols (typically containing branched or unbranched C8-C40 aliphatic moieties) with carboxylic acid groups (e.g., acrylic acid, methacrylic acid), unsaturated cyclic anhydride monomers (e.g., maleic anhydride, itaconic anhydride, citraconic anhydride), monoethylenically unsaturated monoisocyanates (e.g., α,α-dimethyl-m-isopropenyl benzyl isocyanate), or ethylenically unsaturated monomers containing hydroxyl groups (e.g., vinyl alcohol, allyl alcohol). The polyethoxylated and / or polypropoxylated aliphatic alcohols are ethylene oxide and / or propylene oxide adducts of monoalcohols containing C8-C40 aliphatic moieties.Non-limiting examples of alcohols containing a C8-C40 aliphatic moiety include capryl alcohol, isooctyl alcohol (2-ethylhexanol), pelargonic alcohol (1-nonanol), decyl alcohol, lauryl alcohol, myristyl alcohol, cetyl alcohol, cetearyl alcohol (mixture of C16-C18 monoalcohols), stearyl alcohol, isostearyl alcohol, elaidyl alcohol, oleyl alcohol, arachidyl alcohol, behenyl alcohol, lignoceryl alcohol, ceryl alcohol, montanyl alcohol, melissyl, lacceryl alcohol, geddyl alcohol, and C2-C20 alkyl substituted phenols (e.g., nonylphenol).

[0072] Exemplary HASE polymers are disclosed in U.S. Patent Nos. 3,657,175, 4,384,096, 4,464,524, 4,801,671, and 5,292,843. In addition, an extensive 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 sold by Rohm & Haas under the trade names Aculyn® 22 (INCI Name: Acrylates / Steareth-20 Methacrylate Copolymer), Aculyn® 44 (INCI Name: PEG-150 / Decyl Alcohol / SMDI Copolymer), Aculyn 46® (INCI Name: PEG-150 Stearyl Alcohol / SMDI Copolymer), and Aculyn® 88 (INCI Name: Acrylates / Steareth-20 Methacrylate Crosspolymer), and by Lubrizol Advanced Materials, Inc. under the trade name Novethix™ L-10 (INCI Name: Acrylates / Beheneth-25 Methacrylate Copolymer).

[0073] In another embodiment, acid-swellable associative polymers can be used as rheology modifiers. Such polymers generally have cationic and associative properties. These polymers are free radical addition polymers polymerized from a monomer mixture containing acid-sensitive amino-substituted hydrophilic monomers (e.g., dialkylaminoalkyl (meth)acrylates or (meth)acrylamides), associative monomers (as defined above), lower alkyl (meth)acrylates, or other free radical polymerizable comonomers selected from hydroxyalkyl esters of (meth)acrylic acid, 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)acrylic acid, polypropylene glycol esters of (meth)acrylic acid, polyethylene glycol / polypropylene glycol esters of (meth)acrylic acid, and combinations thereof. These polymers can be optionally crosslinked. Acid-sensitive means that the amino substituents become cationic at low pH values, typically in the range of 0.5 to 6.5. Exemplary acid-swellable associative polymers are commercially available from Nouryon under the tradename Structure® Plus (INCI name: Acrylates / Aminoacrylates / C10-C30 Alkyl PEG-20 Itaconate) and from Lubrizol Advanced Materials, Inc. under the tradename Carbopol® Aqua CC (INCI name: Polyacrylate-1 Crosspolymer). In one embodiment, the acid-swellable polymer is a copolymer of one or more C1-C5 alkyl esters of (meth)acrylic acid, C1-C4 dialkylamino C1-C6 alkyl methacrylates, PEG / PPG-30 / 5 allyl ethers, PEG 20-25 C10-C30 alkyl ether methacrylates, hydroxy C2-C6 alkyl methacrylates crosslinked with ethylene glycol dimethacrylate. Other useful acid-swellable associative polymers are disclosed in U.S. Pat. No. 7,378,479.

[0074] Hydrophobically modified alkoxylated methyl glucosides, such as PEG-120 methyl glucose dioleate, PEG-120 methyl glucose trioleate, and PEG-20 methyl glucose sesquistearate, available under the trade names Glucamate® DOE-120, Glucamate™ LT, Glucamate™ VLT, and Glucamate™ SSE-20, respectively, from Lubrizol Advanced Materials, Inc., are also suitable as rheology modifiers.

[0075] Polysaccharides obtained from the exudates of trees and shrubs, such as gum arabic, gum ghatti, and gum tragacanth, as well as pectin; seaweed extracts such as alginates and carrageenans (e.g., lambda, kappa, iota, and salts thereof); algae extracts such as agar; microbial polysaccharides such as xanthan, gellan, and wellan; cellulose ethers such as ethylhexyl ethyl cellulose, hydroxybutyl methyl cellulose, hydroxyethyl methyl cellulose, hydroxypropyl methyl cellulose, methyl cellulose, carboxymethyl cellulose, hydroxyethyl cellulose, and hydroxypropyl cellulose ether; polygalactomannans such as fenugreek gum, cassia gum, locust bean gum, tara gum, and guar gum; starches such as corn starch, tapioca starch, rice starch, wheat starch, potato starch, and sorghum starch can also be used as suitable rheology modifiers in the compositions herein. A particularly suitable rheology modifier used as a thickener is diutan gum.

[0076] The rheology modifiers may be used alone or in combination and may be present in the composition at 0.001 to 50% by weight, based on the total weight of the composition, on an active material basis, for example at least 0.1% by weight, or at least 1% by weight, for example up to 20% by weight, or up to 10% by weight, or up to 3% by weight.

[0077] The mascara composition may include a rheology modifier that is a film former. The film former is dissolved in at least one solvent (such as, for example, water and / or an organic solvent) in the mascara composition, and after the mascara composition is applied to the hair, the at least one solvent evaporates, absorbs, and / or dissipates on the hair, and the film former leaves a film on the hair. The use of a film former can improve the wear of the mascara and impart transfer resistance to the mascara. Film formers are well known in the art and can be any that are cosmetically acceptable for use around the eyes. Examples of useful film formers include natural waxes, polymers such as acrylic acid copolymers, polyethylene polymers, and copolymers of polyvinylpyrrolidone (PVP), ethylene vinyl acetate, dimethicone gum, and resins such as shellac, polyterpene, and various silicone resins such as trimethylsiloxysilicate. The film former can be PVP, acrylic acid copolymers, and polyurethanes. Suitable film formers include, but are not limited to, polyurethanes such as Avalure™ UR450 polymer. In one embodiment, the film formers may be used in an amount ranging 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.

[0078] Additionally, the cosmetic composition may contain other active ingredients such as vitamins, minerals, proteins, peptides, fatty acids, antioxidants, anti-inflammatory agents, darkening agents, and / or mixtures thereof. In particular, the cosmetic composition may contain other active ingredients for promoting hair growth and / or preventing hair loss. Non-limiting examples suitable in the context of the present invention are Growth Oleoactif® marketed by Hallstar [INCI: Helianthus Annuus (Sunflower) Seed Oil (and) Polyglyceryl-3 Diisostearate (and) Carthamus Tinctorius (Safflower) (and) Roselle (Hibiscus Sabdariffa) Flower Extract]; Widelash™ marketed by Sederma [INCI: Glycerin, Water (Aqua), Panthenol, Biotinoyl Tripeptide-1]; SymPeptide® Xlash marketed by Symrise [INCI: Glycerin (and) Aqua (and) Myristoyl Pentapeptide-17], SymLash® 1631 [INCI: Pentylene Glycol (and) Isochrysis Galbana (Isochrysis Galbana Extract]; Anargy™, available from Lipotrue [INCI: Water (and) Butylene Glycol (and) Oligopeptide-2 (and) Nicotiana Benthamiana Hexapeptide-40 sh-polypeptide-9 (and) Nicotiana Benthamiana Hexapeptide-40 sh-polypeptide-86]; Capixyl™, available from Lucas Meyer [INCI: Butylene Glycol (and) Aqua (and) Dextran (and) Acetyl Tetrapeptide-3 (and) Trifolium Pratense (Clover) Flower Extract]; Nano lashes, available from Nanovetores [INCI: Water, Simmondsia Chinensis Seed Extract, Polysorbate 20, Hydroxypropyl Guar, Sodium Benzoate, Potassium Sorbate];SpecPed SC-MH16 [INCI: Water, Glycerin, Myristoyl Hexapeptide-16, Caprylyl Glycol, Ethylhexylglycerin], Myristoyl Pentapeptide-17 [INCI: Myristoyl Pentapeptide-17], SpecPed® BT1 [INCI: Biotinoyl Tripeptide-1], SpecPed® Lash LD [INCI: Biotinoyl Tripeptide-1, Glycerin, Water, Caprylyl Glycol and Ethylhexylglycerin, Panthenol, Toluene], SpecPed® Benzyl Alcohol ... Rehalose], SpecPed® MP17P [INCI: Myristoyl Pentapeptide-17]; Procapil™ marketed by Sederma [INCI: Butylene Glycol (and) Aqua (and) PPG-26-Buteth-26 (and) PEG-40 Hydrogenated Castor Oil (and) Apigenin (and) Oleanolic Acid (and) [Biotinoyl Tripeptide-1]; AnaGain™ marketed by Mibelle [INCI: Pea (Pisum Sativum (Pea) Sprout Extract (and) Isomalt (and) Water], PhytoCellTec™ Malus Domestica Hair [INCI: Apple (Malus Domestica) Fruit Cell Culture Extract (and) Xanthan Gum (and) Glycerin (and) Lecithin (and) Phenoxyethanol (and) Aqua / Water], Santenergy™ [INCI: Bioflavonoids (and) Pentylene Glycol (and) Alcohol (and) Water], RootBioTec™ HW [INCI: Basil (Ocimum Basilicum) Hairy Root Culture Extract (and) Alcohol (and) Water / Water], RootBioTec™ HO [INCI: Basil Hairy Root Culture Extract (and) Sunflower (Helianthus Annuus) Seed Oil (and) Cocos Nucifera (Coconut) Oil; Vytrus Commercially available by Biotech, [INCI: Turmeric (Curcuma longa) Callus Culture Conditioned Medium (and)] Water];marketed by Solabia, [INCI: Glycerin (and) Water (and) Nasturtium Officinale Extract (and) Tropaeolum Majus Extract]; marketed by Peptron, Alotide™, [INCI: Copper Ascorbyl Phosphate Succinyl Tripeptide-34]; marketed by Ichimaru Pharcos, BURGEON-UP, [INCI: Water (and) Alcohol (and) Nasturtium Leaf / Stem Extract]; marketed by Exysmol, Capalgin®, [INCI: Chondrus Crispus Extract]; marketed by BASF, Dermosaccharides® GY, [INCI: Water (and) Glycerin (and) Glycogen (and) Phenoxyethanol (and) Methylparaben], Trichogen™ VEG LS 9922, [INCI: Water (and) Panax Ginseng (and) Ginseng (Ginseng) Root Extract (and) Arginine (and) Acetyl Tyrosine (and) Burdock (Arctium Majus) Root Extract (and) Hydrolyzed Soy Protein (and) Polyquaternium-11 (and) PEG-12 Dimethicone (and) Calcium Pantothenate (and) Zinc Gluconate (and) Niacinamide (and) Ornithine HCl (and) Citrulline (and) Glucosamine HCl (and) Biotin]; Follicusan™ DP marketed by CLR [INCI: Water (and) Denat. Alcohol (and) Panthenyl Ethyl Ether (and) Inositol (and) Milk Proteins (and) Lactose (and) Acetyl Cysteine ​​(and) Acetyl Methionine (and) Sodium Citrate (and) Citric Acid]; Hairline® marketed by Greentech [INCI: Propanediol (and) Water (and) Lindera Strychnifolia) root extract];Kerascalp™ [INCI: Propanediol (and) Glycerin (and) Phyllanthus emblica Fruit Extract], marketed by Provital, Baicapil™ [INCI: Propanediol (and) Water (and) Arginine (and) Lactic Acid (and) Glycine Soja (Soybean) Germ Extract (and) Triticum Vulgare (Wheat) Germ Extract (and) Scutellaria Baicalensis Root Extract (and) Sodium Benzoate (and) Gluconolactone (and) Calcium Gluconate], marketed by Seppic; BIOENERGIZER™ P BG PF [INCI: Water / Water, Butylene Glycol, Panthenol, Propylene Glycol, Pelvetia Canaliculata Extract, Laminaria Digitata Extract, Protectagen™ (INCI: Water (and) Glycerin (and) Hydrolyzed Rice Protein) marketed by Ashland; Redensyl® (INCI: Glycerin (and) Water (and) Sodium Metabisulfite (and) European Larch (Larix Europaea) Extract (and) Glycine (and) Zinc Chloride (and) Tea Plant (Camellia Sinensis) Leaf Extract) marketed by Givaudan; GANOTHER® (INCI: Glycerin (and) Water (and) Ganoderma Lucidum) Mycelium Ferment Filtrate) marketed by B&G; Hairgenyl® (INCI: Saccharomyces Cerevisiae Extract) marketed by Silab, Anageline® (INCI: Hydrolyzed Lupin Protein) marketed by Silab;Hairdian AP [INCI: Propanediol (and) Thuja Orientalis Extract (and) Ginger Root Extract (Zingiber Officinale) Trifolium Pratense (Clover) Leaf Extract (and) Artemisia Argyi Leaf Extract] marketed by Shanghai GREAF Biotech;

[0079] The cosmetic composition may also include an ingredient selected from the group consisting of Minoxidil® (Rogaine), Finasteride® (Propecia), and Dutasteride® (Avodart).

[0080] Purpose The present invention is based on the discovery of surprising properties of a plant extract from a plant of the genus Myrothamnus. In particular, these properties render the plant extract suitable for cosmetic non-therapeutic applications. The cosmetic non-therapeutic applications have the purpose of improving or maintaining an aesthetic appearance, and in particular, the present invention relates to the use of the plant extract for improving or maintaining the aesthetic appearance of hair, in particular for improving or maintaining the aesthetic appearance of eyelashes and / or eyebrows.

[0081] It has been discovered that plant extracts from plants of the genus Myrothamnus can stimulate the proliferation of hair follicle cells, i.e., stimulate the production of new hairs and promote hair growth. It has also been discovered that these extracts can stimulate the production of new blood vessels in endothelial cells and stimulate the proliferation of keratinocyte cells, thereby promoting hair growth and strength. It has also been discovered that the extracts increase the expression of collagen XVII and α6β4 integrin in hair follicles, thereby improving hair anchorage. As a result, the plant extracts of the present invention can promote hair growth, prevent hair loss, and / or increase hair thickness.

[0082] The present invention provides the use of a plant extract of Myrothamnus sp. to promote hair growth. The term "promoting hair growth" means stimulating or enhancing hair growth, and is intended to improve hair density (i.e., per cm of skin). 2 The term refers to increasing the number of hairs per unit area, i.e., growing in a given area of ​​the human body. Optionally or additionally, the term also refers to increasing the length of the hair fiber, i.e., increasing the length of the hair in a given area for a given period of time, compared to the length in the absence of the Myrothamnus sp. extract for the same period of time.

[0083] Hair density may be increased by at least 10%, or at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 100%, at least 200%, or more when an extract of Myrothamnus spp. is used, compared to the density before use of the extract of Myrothamnus spp.

[0084] Hair length, when the length is of average length, may increase by at least 10%, or at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 100%, at least 200% or more when an extract of Myrothamnus spp. is used, compared to the average length when no extract of Myrothamnus spp. is used for the same period of time.

[0085] The present invention provides the use of a plant extract of Myrothamnus species for promoting hair growth in eyelashes and eyebrows.

[0086] The present invention provides the use of a plant extract of Myrothamnus species for lengthening eyelash hair.

[0087] The present invention provides the use of a plant extract of Myrothamnus species for increasing eyelash hair density.

[0088] The present invention provides the use of a plant extract of Myrothamnus species for lengthening eyebrow hair.

[0089] The present invention provides the use of a plant extract of Myrothamnus species for increasing eyebrow hair density.

[0090] The present invention provides a plant extract of Myrothamnus species for preventing hair loss. Hair loss can be normal average daily hair loss or age-related hair loss. Hair loss can be caused by mechanical stress, such as caused by styling hair, or by rubbing with hands or fingers, especially for eyelashes and eyebrows. In particular, the present invention provides the use of a plant extract of Myrothamnus species for preventing hair loss in eyelashes and eyebrows.

[0091] The term "prevention" or "preventing" as used herein refers to the ability of the Myrothamnus species plant extract to prevent, slow or hinder hair loss. The term "prevention" may be interchangeable with the term "reduction", i.e., refers to the ability of the plant extract to reduce the amount of hair lost.

[0092] The present invention provides the use of a plant extract of Myrothamnus species to improve hair hold, so that the hair becomes more resilient, for example against mechanical stress.

[0093] The present invention provides a plant extract of Myrothamnus species for increasing hair thickness. The term "thickness" when used to describe hair refers to the average diameter or cross-sectional area of ​​hair. In certain embodiments, the hair thickness increases by at least 10%, or at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 100%, or at least 200% when Myrothamnus extract is used, compared to the thickness before the use of Myrothamnus extract.

[0094] It has been shown that the plant extract of Myrothamnus species can induce the proliferation of hair follicle dermal papilla cells and keratinocyte cells, increase blood flow in hair follicles, and increase the synthesis of XVII collagen and α6β4 integrin in hair follicles.Therefore, the present invention also relates to the use of the plant extract of Myrothamnus species for promoting hair growth and / or preventing hair loss and / or increasing hair thickness by inducing the proliferation of hair follicle dermal papilla cells, inducing the proliferation of keratinocyte cells and / or associated keratin for hair formation, increasing blood flow in hair follicles, and increasing the synthesis of XVII collagen and α6β4 integrin in hair follicles.

[0095] In one aspect, the present invention provides a method for promoting hair growth and / or preventing hair loss and / or increasing hair thickness, comprising administering to a subject at least one Myrothamnus species plant extract as an active ingredient. The subject is preferably a mammal, more preferably a human. Preferably, the method is cosmetic and non-therapeutic.

[0096] In the above uses and methods, plant extracts may be contained in the compositions as described above.

[0097] The plant extract is preferably administered by topical application. As used herein, "topical application" means that the plant extract (or a composition comprising the plant extract) is brought into contact with mammalian keratinous tissue, such as hair-bearing skin. In particular, the skin is hair-bearing skin, such as scalp skin, facial skin, such as mustache or beard, eyebrows, and eyelash skin. More specifically, the extract or composition is applied to the eyelash line and / or eyebrows, preferably to the eyelash line.

[0098] For the above-described methods of the present invention, the frequency of application or administration can vary widely depending on the needs of each subject, but the recommended application 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, and even more preferably once a day.

[0099] The invention also extends to the combination 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.

[0100] The improvement or maintenance of the aesthetic appearance of hair provided by the above uses and methods can be applied to both healthy subjects and subjects with diseases and / or disorders of skin, hair, nails and / or mucous membranes.Therefore, the methods of the present invention can be applied to subjects that do not suffer from alopecia or subjects that suffer from catagen alopecia, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium or cicatricial alopecia.In addition, the use of the present invention is limited to subjects that do not suffer from alopecia or subjects that suffer from catagen alopecia, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium or cicatricial alopecia.

[0101] In another aspect, a plant extract of Mirothamnus species is provided for use as a medicine.In particular, a plant extract of Mirothamnus species is provided for use in the treatment of hair loss or thinning-related conditions, such as but not limited to alopecia (including catagen alopecia, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium, cicatricial alopecia).In another aspect, a method for treating hair loss or thinning-related conditions, such as but not limited to alopecia (including catagen alopecia, androgenetic alopecia, alopecia areata, alopecia universalis, trichotillomania, telogen effluvium, cicatricial alopecia), is provided, comprising administering a therapeutically effective amount of a plant extract of Mirothamnus species to a subject.

[0102] The invention is further described in the following non-limiting examples. EXAMPLES

[0103] Example 1 Extracts obtained by conventional aqueous extraction Conventional extraction was performed on dried Myrothamnus flabellifolia stems with leaves chopped into approximately 1.27-0.22 mm pieces. The plant material was subjected to extraction with water in a ratio of 1:10 (dried plant material:water, w / v). Extraction was carried out 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% glycerin in a ratio of 1:5 (extract:glycerin, w / w) and the resulting composition was mixed to form a stock solution.

[0104] Example 2 Extracts obtained by subcritical water extraction Subcritical water extraction (SWE) was performed on dried Myrothamnus flabellifolia stems with leaves ground to a coarse powder of approximately 1.27-0.25 mm. The powder was extracted with subcritical water in a ratio of 1:20 (powder:water, w / v). The subcritical water was kept at a temperature of 150 °C under a pressure ranging from 10-11 MPa for 1 h. The resulting solution was then allowed to cool to room temperature (25 °C) and filtered through a 10 μm filter. The filtered solution was the plant extract, which was then diluted with glycerin in a ratio of 1:5 (extract:glycerin, w / w). The resulting composition was mixed to form a stock solution.

[0105] Example 3 Composition analysis of extracts The stock solutions of Examples 1 and 2 were evaluated for differences in phytochemicals. Total polyphenol content (TPC) was analyzed along with the identity of individual phytochemicals.

[0106] Total polyphenol content was determined by the Folin-Ciocalteu method. Briefly, 2 mL of extract was mixed with 5 mL of distilled water, then 1 mL of Folin-Ciocalteu reagent was added and mixed well for 3 min, after which the reaction was stopped by adding 5 mL of 10% sodium carbonate solution. After 30 min at room temperature, the absorbance was measured at 760 nm and 850 nm using a spectrophotometer. A calibration curve was obtained using gallic acid as a standard.

[0107] Liquid chromatography and tandem mass spectrometry LC / MS-MS was used to identify more specific phytochemicals. Ten microliters (10 μL) of the extract was mixed with 100 μL of 10 mM HCL and 890 μL of ultrapure water in a 2 mL LC-MS vial to dilute the sample 100-fold. The injection volume was 2 μL. The mobile phase used for this analysis contained 0.1% formic acid in acetonitrile (solvent A) and 0.1% formic acid in ultrapure water (solvent B). The flow rate was 0.2 mL / min with a column temperature of 40 °C.

[0108] The total polyphenol content of the aqueous plant extract of Example 1 was 652 mg GAE / L by 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. The ND designation indicates a compound not detected in the sample. [Table 1]

[0109] Example 4 In vitro proliferation assay in hair follicle dermal papilla cells Myrothamnus flabellifolia stock solutions ("Test Products") obtained as described in Examples 1 and 2 were dissolved at 0.5, 1, 2, 2.5, and 5% (v / v) in papilla cell growth medium supplemented with growth supplements and filtered through 0.2 µm syringe filters.

[0110] Hair follicle dermal papilla cells (HFDPCs) derived from adult human scalp were cultured at 3 × 10 per mL in papilla cell growth medium for 5 days at 37 °C in 5% CO2 humidified air. 4 The cells were allowed to grow to density after which they were split and then seeded for treatment.

[0111] HFDPCs were seeded into cultures at a density of 45000 cells / mL in 96-well Cell Carrier black plates (PerkinElmer) in culture medium. After 24 hours of incubation at 37°C in 5% CO2 humidified air, the medium was removed and dilutions of the test products and vehicle control were added in fresh culture medium. Cells treated with medium only were used as negative control. Fetal bovine serum (FBS) was used as positive control.

[0112] After 24 hours of incubation, live and dead cells were quantified using the LIVE / DEAD® Viability / Cytotoxicity Kit (Thermo Fisher Scientific), which distinguishes live and dead cells by simultaneous staining.

[0113] Test products were assayed in three independent experiments with four replicates in each experiment. Table 2 shows the percentage of live and dead HFDPCs normalized to the total amount of cells (total) in the negative control according to the following formula:

number

number

[0114] The results show that the M. flabellifolia extract increases the number of live cells and decreases the number of dead cells compared to the negative control (untreated cells). Subsequently, the extract has a proliferative effect on HFDPCs.

[0115] Example 5 In vitro proliferation assay in keratinocyte cells Keratinocytes express keratin, the most abundant protein in hair fibers, and contribute to the de novo formation of hair.

[0116] Myrothamnus flabellifolia stock solutions were dissolved at 0.5, 1, 2, 2.5, and 5% (v / v) in keratinocyte growth medium supplemented with Mix C39016-CaCl2 solution and filtered through a 0.2 μm syringe filter.

[0117] Human epidermal keratinocyte cells (HEK) were cultured at 4500 cells / cm in keratinocyte growth medium at 37°C in 5% CO2 humidified air for 7 days. 2 After this, they were split and then seeded for treatment.

[0118] HEKs were seeded in 96-well Cell Carrier black plates (PerkinElmer) in culture medium at a density of 70000 cells per mL of culture medium. After 24 hours of incubation at 37°C in 5% CO2 humidified air, the medium was removed and dilutions of the test products were added in fresh culture medium. Cells treated with medium only were used as negative control. Epidermal growth factor (EGF) was used as positive control. After 24 hours of incubation, live and dead cells were quantified with PrestoBlue® cell viability reagent.

[0119] HEK quantification was quantified by fluorescence by using the cell permeable resazurin-based solution PrestoBlue® from Thermo Fisher Scientific.

[0120] Stock solutions of Examples 1 and 2 were assayed in three independent experiments with four replicates in each experiment. result Table 3 shows the cell proliferation values ​​of HEK cells normalized to the negative control. [Table 3] The results demonstrate that M. flabellifolia extract increases keratinocyte cell proliferation.

[0121] Example 6 In vitro study of new blood vessel formation in human umbilical vein endothelial cells (HUVEC) Hair follicles receive nutrition from the bloodstream of hair-bearing skin. Optimal irrigation of hair follicles increases nutrition, stimulates new hair production, and improves hair growth and strength. Compounds that can generate new blood vessels in hair follicles are good candidates for cosmetic treatment of eyelash growth and strength. In vitro evaluation of new blood vessel formation includes measuring the ability of endothelial cells to form three-dimensional tube-like structures. Human umbilical vein endothelial cells (HUVECs) mediate specific binding with extracellular matrix components to generate differential forces that induce morphological changes and migration of HUVECs, resulting in the formation of tube-like structures. The purpose of this study is to evaluate the efficacy of compounds of the present invention to generate new vascular tubes by measuring the ability of endothelial cells to form three-dimensional tubular structures.

[0122] A 96-well plate was pre-coated with cold extracellular matrix solution (angiogenesis assay kit PromoKine) and incubated at 37°C for 1 hour. After incubation, HUVEC cells were seeded into the pre-coated wells at a density of 15000 cells per well. Immediately, the cells were treated with the stock solution of 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 5ng / ml, and cells treated with culture medium only were used as a basal control. The cells were treated for 4 hours at 37°C in 5% CO2, and then stained with dye (angiogenesis assay kit, ProMoKine) according to the manufacturer's protocol. Briefly, the cells were washed and then dye solution was added. After 30 min of incubation at 37° C. in 5% CO 2 humidified air, fluorescent images were captured using an Operetta® confocal microscope (PerkinElmer, Inc.) and new tube formation was assessed by ImageJ software.

[0123] New vascular tube formation was assessed by the measurement of different parameters in each acquired image. First, ImageJ detected the components of the network, such as junctions, segments, and branches. Junctions are nodes that fit into the branches of tubes, segments are tubes defined by two junctions, and branches correspond to elements defined by only one junction. The software then quantified more complex structures. -Segment Length: Number of pixels in each detected segment -Master Segment Count: The number of segments separated by junctions that have no branch connections. -Number of master junctions: The number of junctions that link at least three master segments Mesh count: Number of elements separated by a segment or master segment

[0124] To consider an increase in new vessel formation, two or more of the parameters being measured need to be increased.

[0125] The results herein are the different measured parameters expressed as percentages normalized by basal conditions. [Table 4] The mean values ​​of the different measured parameters are expressed as percentages relative to basal conditions in HUVEC cells.

[0126] The results confirm that the plant extract of the present invention increases tube-like structure formation over basal conditions in HUVEC cells at the concentrations tested. The results also demonstrate that the increase achieved by the plant extract of the present invention is similar to that of the positive control.

[0127] Example 7 Ex vivo study of collagen XVII and integrin β-4 immunostaining in scalp explants Hair anchorage depends on the adhesion of different cells to the basement membrane area, with hemidesmosomes being the main binding unit. Hemidesmosomes are cell-matrix junctions formed by collagen XVII, two subunits of α6β4 integrin, and CD151. Both collagen XVII and α6β4 integrin directly interact with the basement membrane zone and help to avoid hair loss. Furthermore, collagen XVII proteolysis is associated with hair follicle aging. It has been demonstrated that hair follicles become smaller during aging and often disappear from the skin due to a miniaturization process characterized by collagen XVII proteolysis, leading to hair thinning and hair loss [Matsumura H et al., "Hair follicle aging is driven by transepidermal elimination of stem cells via COL17A1 proteolysis," Science. 2016 Feb 5;351(6273):aad4395]. Compounds capable of increasing collagen XVII and α6β4 integrin are good candidates for the cosmetic treatment of aging, strength and loss of eyelashes and eyebrows. The aim of this study was to evaluate the efficacy of compounds of the extract of the present invention in improving hair retention by measuring the increase of collagen XVII and integrin β-4 (β4) in different regions of hair follicles in scalp explants.

[0128] Scalp explants were prepared and maintained in BEM medium (BIO-EC) at 37°C in a humidified 5% CO2 atmosphere for 8 days. A 2% (w / w) dilution of the stock solution of Example 2 was applied topically to the explants on days 0, 4, and 6. Scalp explants treated with medium only were used as basal controls. On day 8, the explants were harvested and cut into two parts. One half was fixed in a buffered formalin solution and the other half was frozen at -80°C.

[0129] After fixation in buffered formalin for 24 hours, the explants were dehydrated and impregnated with paraffin. The explants were then embedded and sectioned at 5 μm using a microtome (Leica). The sections were mounted on histological slides. Collagen XVII immunostaining was performed with monoclonal anti-collagen XVII antibody (Abcam) for 1 hour at room temperature using the Vectastain Kit Vector Amplification System Avidin / Biotin and revealed by VIP (Vector laboratories, Ref. SK-4600), a substrate for peroxidase that gives a purple stain when oxidized. Immunostaining was assessed by microscopic observation and semi-quantified by image analysis.

[0130] Frozen explants were cut into 7 μm thick sections using a cryostat (Leica). Sections were then mounted on silanized glass slides. Integrin β4 immunostaining was performed on frozen skin sections with monoclonal anti-integrin β4 antibody (Chemicon) for 1 h at room temperature and revealed by AlexaFluor488 (Life technologies). Nuclei were counterstained with propidium iodide. Immunostaining was performed manually and semi-quantified by image analysis.

[0131] Microscopic observations of collagen XVII and integrin β4 were imaged with a camera (Olympus) and analyzed with CellSens software. Different regions of the hair follicle were studied by defining the following regions of interest: dermal-epidermal junction, infundibulum, upper root sheath, bulge, lower root sheath, and hair bulb. In each of these regions, collagen XVII and integrin β4 were quantified.

[0132] Results herein are the percentage increase in collagen XVII and integrin β4 in the hair follicle region relative to basal controls. [Table 5] The percentage of increase in collagen XVII and integrin β4 in different regions of the hair follicle relative to the basal control ( *p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001)

[0133] The results demonstrate that the plant extract of the present invention stimulates hair anchorage by increasing the expression of collagen XVII and integrin β4 along the hair follicle.

[0134] Example 8 Mascara for eyelashes An eyelash mascara according to the present invention was prepared as follows. The ingredients of Phase A were mixed under stirring while heating at 85° C. and then dispersed in the ingredients of Phase B. Subsequently, the ingredients of Phase D were added. The ingredients of Phase E were mixed with heating at 90° C. An emulsion was made by slowly adding Phase E to the mixture of Phases A-D and finally adding Phase F. [Table 6]

[0135] Example 9 Eyelash serum The ingredients of Phase A were mixed together. Then the ingredients from Phase B were added with stirring. The mixture was neutralized with the ingredients of Phase C, and finally Component D was added. [Table 7] Example 10 In vivo study to evaluate the increase in eyelash length of Myrothamnus species extract after chronic application in Caucasian female volunteers The study will be carried out for 56 days. The volunteer group consisted of 31 Caucasian female volunteers aged 25-50 years. All individuals in the study applied the active and placebo products in a half-face design, with the active serum of Example 9 and the mascara of Example 8 applied to the eyelashes on one half, and the placebo serum and mascara (with the same formulation but without Myrothamnus species extract) on the other half. The serum was applied to the base of the eyelashes in the morning and at night, and the mascara was applied only in the morning for 56 days. During the treatment period, all volunteers used a cleanser product (Eucerin® DermatoCLEAN from Beiesdorf) only in the evening to remove the serum and mascara applied in the morning, before applying the serum at night. The subjects served as their own reference and the results obtained on the 56th day were compared with those obtained at the first time. Furthermore, the results obtained with the active formulation were compared with those obtained with the placebo serum. The effectiveness of the compositions was evaluated by measuring eyelash length.

[0136] Before and after 56 days of product use, side macro photographs were taken of the face from 20 degrees using the software CameraScan (Orion Concept). The photographs were taken with the eyes closed so that the upper eyelid lashes were lying on a white patch. Acquisition was done with parallel polarized light and the images were analyzed by Image J (National Institutes of Health) software. The photographed lashes were divided into five sections and the longest lashes of each section were selected for measurement. The results are shown in Table 8. [Table 8] Increase in mean eyelash length after 56 days of product application Statistical significance vs. baseline: calculated using unpaired Student's t-test vs. baseline * p<0.05

[0137] The results demonstrate that after 56 days of application of the serum and mascara of the present invention, there is a statistically significant increase in the average length of the eyelashes compared to the starting point. Furthermore, the increase in eyelash length is greater with the active formulation than with the placebo formulation.

[0138] Example 12 In vivo study to evaluate the improvement of brow density of Myrothamnus extract after long-term application in Caucasian female volunteers The study will be carried out for 56 days. The volunteer group consisted of 28 Caucasian female volunteers aged 25-50 years. All individuals in the study applied the active serum of Example 9 and the placebo serum (having the same ingredients except for Myrothamnus species) in a half-face design. The serums were applied to the eyebrows twice a day (morning and night) during the 56-day study. In addition, during the treatment period, all volunteers used the cleanser product Eucerin® DermatoCLEAN manufactured by Beiesdorf only in the evening to remove the serum applied in the morning, before applying the serum in the evening. The subjects served as their own reference and the results obtained on the 56th day were compared with those obtained at the first time. In addition, the results obtained with the active serum were compared with those obtained with the placebo serum.

[0139] The effectiveness of the composition was evaluated by brow density measurement. Before and after 56 days of product use, side macro photographs were taken of the face from 35 degrees using the software CameraScan (Orion Concept). The images obtained were analyzed with Image J software. The brow area was selected and converted into a binary image. The white pixels were counted and the following results were obtained: [Table 9] Increase in mean brow density after 56 days of product application Statistical significance versus baseline: calculated using unpaired Student's t-test versus baseline * p<0.1 The results demonstrate that after 56 days of application of the serum of the present invention, there is a statistically significant increase in eyebrow density compared to the starting point.

[0140] Example 13 An in vivo study to assess cilia curvature angle after long-term application in Caucasian female volunteers The study will be conducted for 56 days. A volunteer panel of 31 Caucasian female volunteers aged 25-50 years was included in the study. All individuals in the study applied the active and placebo products in a half-face design, with the active serum and mascara applied to the eyelashes on one half and the placebo serum and mascara applied to the other half. The serum was applied to the base of the eyelashes once in the morning and once in the evening, and the mascara was applied only in the morning for 56 days. During the treatment period, all volunteers used a cleanser product (Eucerin® DermatoCLEAN from Beiesdorf) only in the evening to remove the serum and mascara applied in the morning, before applying the serum at night. The active mascara and serum are described in Examples 8 and 9, respectively. The placebo mascara and serum had the same ingredients as the active mascara and serum, except for the Myrothamnus species extract. The subjects served as their own baseline and the results obtained on day 56 were compared with those obtained at the initial time. The effectiveness of the composition was evaluated by determining the change in eyelash curvature angle using the following formula:

number

[0141] Before (initial) and after 56 days of product use, lateral macrophotographs of the face were taken from 90 degrees using the software CameraScan (Orion Concept). Acquisition was performed with parallel polarized light and images were analyzed by Image J (National Institutes of Health) software. The topmost eyelash was selected for each image and the curvature angle was measured at the midpoint of the eyelash. [Table 10] Statistical significance of change in mean lash curvature angle after 56 days of product application versus baseline: calculated using unpaired Student's t-test versus baseline * p<0.001

[0142] The results demonstrate that after 56 days of application of the composition of the present invention, there is a statistically significant decrease in the average eyelash curvature angle compared to the starting point.

[0143] Example 14 In vivo study for the assessment of eyelash and eyebrow hair loss The study will be conducted for 28 days. Thirty-one Caucasian female volunteers between the ages of 25 and 50 were included in the study. All individuals in the study applied the active and placebo products in a half-face design. For the eyelashes, one half of the eyelashes was used with the active serum and mascara, and the other half with the placebo serum and mascara. The serum was applied to the base of the eyelashes once in the morning and once at night, and the mascara was applied only once in the morning for 28 days. The eyebrows were treated with the serum using the same pattern of active and placebo half design. The serum was applied to the eyebrows in the morning and at night for 28 days. During the treatment period, all volunteers used a cleanser product only at night to remove the serum, and applied mascara before applying the serum in the morning and at night. The active serum and mascara are described in Examples 8 and 9. Furthermore, the results obtained with the active formulation were compared to those obtained with the placebo formulation. The effectiveness of the compositions was assessed by the total number of hairs lost from the eyelashes and eyebrows.

[0144] The loss of eyelashes and eyebrows was counted daily, and each eyebrow and eyelash was washed separately to count the number of hairs lost for the active product and the placebo product.The results shown in Table 11 correspond to the total number of hairs lost for all volunteers according to each condition and location.

[0145] [Table 11]

[0146] The results demonstrate that after 28 days of application of the serum and mascara of the present invention, there is a reduction in the number of eyelash and eyebrow hairs lost when the active treatment is applied compared to a placebo.

Claims

1. A composition comprising a plant extract of Myrothamnus species for use in promoting hair growth and / or preventing hair loss and / or increasing hair thickness, wherein the extract is obtained in a solid / liquid extraction using subcritical water as the extraction solvent.

2. 1. A composition comprising a plant extract of Myrothamnus species for use in lengthening eyelashes, said extract being obtained in a solid / liquid extraction using subcritical water as the extraction solvent.

3. 1. A composition comprising a plant extract of Myrothamnus species for use in increasing the density of eyebrow hair, said extract being obtained in a solid / liquid extraction using subcritical water as the extraction solvent.

4. 1. A composition comprising a plant extract of Myrothamnus species for use in improving hair retention, said extract being obtained in a solid / liquid extraction using subcritical water as the extraction solvent.

5. The composition of claim 1 , wherein the hair is an eyelash or an eyebrow hair.

6. The composition described in claim 4, wherein the hair is eyelash or eyebrow hair.

7. The composition according to any one of claims 1 to 4, wherein the extract is obtained from the aerial parts of Myrothamnus species.

8. The composition according to any one of claims 1 to 4, wherein the plant extract is a plant extract of the species Myrothamnus flabellifolia.

9. The composition of any one of claims 1 to 4, wherein the plant extract comprises mykelianin and kaempferol-3-O-glucuronide; or mykelianin, kaempferol-3-O-glucuronide, and trehalose; or mykelianin, kaempferol-3-O-glucuronide, trehalose, and luteolin-7-O-glucuronide; or mykelianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and arbutin.

10. The plant extract may comprise: mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, and isorhamnetin-3-O-glucoside; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, and naringenin; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, and The composition of any one of claims 1 to 4, comprising: piceid; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, piceid, and coniferaldehyde; or mychellianin, kaempferol-3-O-glucuronide, trehalose, luteolin-7-O-glucuronide, isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

11. 5. The composition according to any one of claims 1 to 4, which promotes hair growth and / or increases hair thickness and / or prevents hair loss by inducing proliferation of hair follicle dermal papilla cells, inducing proliferation of keratinocyte cells and / or associated keratin for hair formation, increasing blood flow within the hair follicle, and / or increasing synthesis of collagen XVII and α6β4 integrin in the hair follicle.

12. The composition according to any one of claims 1 to 4, wherein the extract is an aqueous extract.

13. 13. The composition of claim 12, wherein the extract is in a composition further comprising a water-miscible organic solvent, the water-miscible organic solvent being a polyol.

14. 14. The composition of claim 13, wherein the weight ratio of the aqueous extract to the organic miscible solvent is from 1:1 to 1:

10.

15. 5. The composition according to any one of claims 1 to 4, wherein the extract is in a cosmetic composition comprising at least one cosmetically acceptable excipient or ingredient, the cosmetic composition being selected from the group consisting of a lotion, a shampoo, a serum, or a mascara composition.

16. 16. The composition according to claim 15, wherein the composition is in the form of a serum for the eyebrows or eyelashes, or a mascara composition for the eyelashes.

17. The composition of claim 16, wherein the concentration of the extract is 0.5 to 5% (w / w).

18. 1. A process for obtaining a plant extract from Myrothamnus species, comprising subjecting plant material of Myrothamnus species to extraction with subcritical water, said extraction comprising: i) contacting the plant material from Myrothamnus species with subcritical water at a temperature of 120-220°C and at a temperature and pressure suitable to maintain the water in a liquid state for at least 10 minutes to form an aqueous plant extract; ii) separating said plant material from said aqueous plant extract.

19. 19. The process of claim 18, wherein the temperature of the subcritical water is from 140°C to 160°C.

20. 19. An aqueous plant extract obtainable by the process according to claim 18.

21. 21. The aqueous plant extract of claim 20, comprising at least one compound selected from the group consisting of isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

22. 22. The aqueous plant extract of claim 21, comprising isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside.

23. 22. The aqueous plant extract of claim 21, wherein the concentration of each of isorhamnetin-3-O-glucoside, naringenin, piceid, coniferaldehyde, and naringenin-7-O-glucoside is at least 0.05 ppm.

24. 21. A composition comprising the aqueous plant extract of claim 20 and a water-miscible organic solvent, wherein the water-miscible organic solvent is a polyol.

25. The composition of claim 24, wherein the polyol is present in an amount of 50 to 90% by weight based on the weight of the total composition.

26. 25. A cosmetic composition comprising the aqueous plant extract of claim 20 or the composition of claim 24, and at least one cosmetically acceptable excipient or ingredient.

27. ​​A cosmetic composition as described in claim 26, comprising the composition as described in claim 24 in an amount of 0.01 to 20% by weight.

28. A mascara composition comprising a plant extract of the Myrothamnus species and at least one wax, said extract being obtained in a solid / liquid extraction using subcritical water as the extraction solvent.