Verbena extract for preventing and / or treating hair loss

Abstract

This invention relates to a verbena extract and compositions containing the verbena extract, which are used in the fields of cosmetics and dermatology, and in the field of hair care, particularly for the prevention and / or treatment of hair loss or for promoting hair growth.

Description

Technical Field

[0001] This invention relates to a verbena ( Verbena officinalis Verbena extract and compositions containing the verbena extract are used in the fields of cosmetics and dermatology, and in the field of hair care, particularly for the prevention and / or treatment of hair loss or for promoting hair growth. Background Technology

[0002] Hair care has always been a subject of research, not only for cosmetic purposes, but also for preventing hair loss and promoting hair regrowth. Many theories attempt to elucidate the causes of hair loss in conditions such as baldness, alopecia areata, and patchy hair loss, attributing it to seborrhea, increased tension in the skull tissues, reduced blood flow, or certain endocrine or neurological disorders.

[0003] Hair follicles are tiny organs embedded in the skin and extending into the subcutaneous tissue, whose primary function is to produce hair shafts. Their distribution is determined during intrauterine growth, and their number is genetically determined. Hair follicles are dynamic structures that produce hair during the tissue's growth and remodeling cycle. This cycle consists of three phases: - Anagen Phase (Early Hair Growth Phase): Cells in the dermal papilla (fibroblasts) send signals to the stem cells of the hair follicle, enabling them to proliferate. These cells transform and surround the dermal papilla to form the hair matrix. They divide and differentiate into keratinocytes, the cells responsible for hair structure. For hair to have a good structure, these keratinocytes require sulfur-containing proteins, vitamin B6, and various minerals such as zinc and magnesium. The duration of this phase determines hair length and depends on the proliferation and differentiation of stromal cells at the base of the hair follicle.

[0004] - Regression phase (mid-hair growth phase): The matrix dies, therefore, the dermal papilla no longer comes into contact with the matrix. There is no longer any exchange between cells. The hair follicle and dermal papilla move towards the epidermis.

[0005] - Telogen effluvium (telogen phase): The cells of the dermal papilla and hair follicle remain intact and inactive. Hair falls out. The cycle must restart in order for new hair to grow.

[0006] The term hair loss refers to the partial or complete loss of hair. Many factors can cause hair loss, including genetics, age, gender, illness, stress, hormonal imbalances, medication side effects, and scarring. Several forms of hair loss can be distinguished: - Hereditary androgenetic alopecia: This is the most common type. Early hair loss occurs in individuals with a genetic predisposition and affects men particularly. It manifests as a progressive and localized reduction in hair volume, sometimes leading to baldness, affecting 50% of men over 50. Androgenetic alopecia, also known as male or female pattern baldness, is a progressive and hereditary form of hair loss. It is primarily caused by the action of androgens on hair follicles. The role of 5-α-reductase in androgenetic alopecia is well-established. This enzyme converts testosterone to dihydrotestosterone (DHT), a more potent hormone. DHT binds to receptors on hair follicles, causing them to shrink and shorten the hair's growth phase (the initial stage of hair growth). This process leads to a gradual reduction in follicle size, resulting in thinner, shorter hair and ultimately, hair loss. Postmenopausal hair loss: This is the most common cause of hair loss in women. In women, hair loss is more diffuse and widespread than in men. Diffuse female pattern baldness is a condition that typically begins around menopause and affects approximately 40% of women over 70. The term "diffuse" indicates that, unlike in men, hair loss affects the entire scalp in a uniform manner.

[0007] - Emotional alopecia, also known as reactive alopecia, or sometimes called acute alopecia, is a form of diffuse hair loss that typically occurs after a stressful event or physiological change. Emotional / reactive alopecia is characterized by an increase in the number of hairs that prematurely enter the telogen (resting) phase of the hair growth cycle (telogen effluvium). It can be associated with chemotherapy, stress, childbirth, significant nutritional deficiencies, iron deficiency, and hormonal imbalances; it involves the simultaneous and diffuse shedding of large amounts of hair. The term emotional / reactive alopecia is generally used to describe hair loss caused by emotional or psychological factors such as intense stress or emotional trauma. Emotional / reactive alopecia specifically includes a form known as telogen effluvium. Telogen effluvium can be a manifestation of this form of hair loss because emotional stress is one of the known triggers for telogen effluvium.

[0008] Telogen effluvium is a form of diffuse hair loss that occurs when hair follicles prematurely enter the telogen phase (the resting phase of the hair growth cycle). This can be triggered by a variety of factors, including emotional stress, hormonal changes, illness, medications, or physiological events such as childbirth. In short, telogen effluvium can be considered a specific form of reactive / emotional hair loss, typically triggered by emotional or physiological factors. Telogen effluvium can be acute or chronic. When hair loss occurs suddenly, rapidly, and diffusely, it is called acute telogen effluvium. Acute telogen effluvium can become chronic and fluctuate for several years, during which time the hair cycle shortens; in this case, it can be called chronic telogen effluvium. When hair loss persists for more than 6 months, telogen effluvium is considered chronic.

[0009] Unlike androgenetic alopecia, which has a genetic origin, reactive / emotional hair loss (as the name suggests) occurs in response to external events (stress, pregnancy, breastfeeding, seasonal changes, fatigue, illness, etc.). Hair loss typically occurs 2 to 3 months after the event. This type of hair loss can affect the entire scalp and is more common in women.

[0010] According to Kubanov et al. (Biomedical & Pharmacology Journal Vol. 10(1), 191-198(2017)), various growth factors (VEGF, KGF, EGF, TGF-β1) contribute to the development of telogen effluvium (or emotional or reactive alopecia), with VEGF having the greatest impact. Analysis showed that women with telogen effluvium had significantly lower VEGF expression in their scalps compared to women without this form of hair loss. This decrease in VEGF was associated with an increased likelihood of developing telogen effluvium. VEGF plays a crucial role in angiogenesis and vascularization, thereby stimulating hair growth and regeneration. Therefore, decreased VEGF expression in hair follicles is a key factor in the development of this form of hair loss.

[0011] - Scarring alopecia: It can be caused by skin problems (tumors, burns, alopecia areata), acute radiation, lupus erythematosus, or parasites (tinea, lichen); - Alopecia areata: It appears to have an autoimmune origin and is characterized by plaque-like lesions of varying sizes appearing in one or more locations; - Congenital hair loss: rare, it corresponds to missing hair roots or abnormal hair (mutation).

[0012] Hair loss, especially androgenetic alopecia, is primarily associated with the disruption of hair renewal, which initially leads to an accelerated cycle frequency, first at the expense of hair quality, and then quantity. The most common phenomenon is a shortened duration of the anagen phase (the initial stage of hair growth) due to the cessation of cell proliferation. The result is premature induction of the catagen phase and a greater number of hair follicles entering the telogen phase, leading to more significant hair loss.

[0013] To date, a variety of products have been developed to combat hair loss, particularly those that allow the induction or stimulation of hair growth. Most products combine several active ingredients that may have beneficial effects on the biological parameters involved in hair loss. Among the most common active ingredients, examples include: vitamins such as vitamins A, E, B5, B6, C, H, and PP; trace elements such as zinc, copper, magnesium, and silicon; protein derivatives such as peptides and sulfur-containing amino acids (e.g., methionine, cysteine, or derivatives); lipophilic or hydrophilic plant-derived essential oils or extracts (this list is not exhaustive); and antifungal agents such as piroctone olamine, undecenoic acid derivatives, and ciclopirox olamine. A chemically synthesized molecule known for its specific action on androgen receptors or 5-α-reductase activity—minoxidil, or 2,4-diamino-6-piperidinylpyrimidine 3-oxide—is now the standard treatment for androgenetic alopecia, preventing hair loss.

[0014] In addition, another alternative aims to stimulate / restart hair regeneration and proposes compositions containing a wide variety of active ingredients in hair regeneration, such as derivatives of 2,4-diaminopyrimidine 3-oxide, for example those described in patent application EP0522964.

[0015] Therefore, despite the many options available today, consumers still need new products that are natural and environmentally friendly, yet just as effective as chemically active substances in promoting hair regeneration.

[0016] verbena ( Verbena officinalis *L.* (Verbenaceae) is a perennial herb native to Europe, commonly known as common verbena. It has been widely naturalized outside its native range, for example, in North America.

[0017] Common verbena is a perennial plant with spindle-shaped, pale yellow roots. It is an erect or ascending plant, often more or less perennial. The stem is slightly hairy, quadrangular, alternately grooved on two opposite sides, branching and then erect. Leaves are opposite, with short petioles or even sessile, obovate to lanceolate, with unequal lobes, said lobes deeply lobed or crenate. Pale blue or pale purple flowers are small, sessile, arranged in long, loose, linear spikes in terminal panicles. Each flower has a slender, green bract at its base, less than 3 mm long (shorter than the calyx). The funnel-shaped corolla includes a prominent tube with nearly flat leaves and five nearly equal lobes. The fruit is a brown, dehiscent capsule composed of four achenes that separate upon maturity. It is a common plant found everywhere: on dikes, gravel, roadsides, especially in villages, on old walls, in uncultivated areas, except in extremely high mountains. Today, it is widely distributed in all temperate regions of the world.

[0018] Verbena officinalis possesses a rich phytochemical composition. The main components of its bioactive secondary metabolites are glycosylated iridoids, including verbenacin (approximately 6% in the dried plant), aucubin, and eugenol (approximately 0.6% in the dried plant), as well as glycosylated phenylpropanoids and caffeic acid derivatives: verbacin (approximately 3% in the dried plant), isoveracin (approximately 0.3% in the dried plant), and eugenol. Furthermore, numerous flavonoids have been identified in verbena extracts, including common plant-based compounds such as kaempferol and luteolin, as well as specific flavonoids such as baicalin and linalool. It should also be noted that phenolic acids are present: derivatives of chlorogenic acid, ferulic acid, protocatechuic acid, rosmarinic acid, and dicaffeoylquinic acid. Finally, the presence of terpenoids such as citral, limonene, eucalyptol, carvone, diterpenoids such as salviaol, salviaol, rosmarinic acid, sesquiterpenoids such as caryophyllene oxide, α-curcumene, and triterpenoids such as ursolic acid and its derivatives was described in common verbena (Kubica P et al., Planta Med 2020; 86: 1241–1257).

[0019] Recent pharmacological studies, both in vitro and in vivo, have attempted to establish important new applications for extracts of common verbena by demonstrating antioxidant, antibacterial, anti-inflammatory, anticancer, neuroprotective, analgesic, or anticonvulsant activities (Kubica P et al., Planta Med 2020; 86: 1241–1257).

[0020] However, to date, there is no written data mentioning that verbena extract can be active in the prevention and / or treatment of hair loss, especially emotional or reactive hair loss (more specifically telogen effluvium), by promoting hair regeneration. Summary of the Invention

[0021] This invention relates to the use of a verbena extract for the prevention and / or treatment of hair loss, and to a cosmetic or dermatological composition for the prevention and / or treatment of hair loss, particularly reactive / emotional alopecia or more particularly telogen effluvium (more particularly by stimulating hair regrowth), said cosmetic or dermatological composition comprising at least one verbena extract and at least one dermatologically or cosmetically acceptable excipient.

[0022] Other aspects of the invention are described in the claims and below. Detailed Implementation

[0023] The inventors unexpectedly discovered that verbena extract can inhibit apoptosis of outer root sheath cells and possesses particularly significant pharmacological activity in stimulating angiogenesis, especially by acting on VEGF targets to restart hair regeneration. Indeed, VEGF (vascular endothelial growth factor) is known to play a crucial role in angiogenesis and vascularization, thereby stimulating hair growth. Decreased VEGF expression in hair follicles is associated with an increased likelihood of developing telogen effluvium (also known as reactive / emotional alopecia).

[0024] In addition, this verbena extract also has pharmacological activity that promotes hair growth, particularly by acting on the JAK / STAT pathway.

[0025] Therefore, the present invention relates to hair loss, particularly reactive / emotional hair loss, preferably telogen effluvium, and advantageously to the prevention and / or treatment of acute telogen effluvium, since verbena extract acts on the biological functions or mechanisms responsible for hair growth.

[0026] definition In this invention, the plant verbena ( Verbena officinalis L.) can be abbreviated as verbena ( Verbena officinalis "Verbena extract" refers to a product extracted from the whole or part of the verbena plant.

[0027] "The above-ground parts of verbena" refers to all the above-ground parts of the plant, namely, the stems, leaves, flowers, fruits, and seeds.

[0028] The term "extraction product" refers to the product obtained after extracting whole or part of a plant using a solvent called an extraction solvent. That is, the product present in the extraction solvent, which may be in concentrated or dried form after partial or complete evaporation of the extraction solvent. It can be a dry extract.

[0029] For the purposes of this document, "dry extract" refers to an extract that contains no extraction solvent or carrier, or contains only trace amounts of such solvent or carrier. Therefore, this dry extract contains only the material obtained from verbena extract.

[0030] In this invention, "about" means that the relevant value may be 10% lower or higher than the specified value, particularly 5%, and especially 1% higher. This definition is particularly applicable when defining the weight content of iridoid compounds (such as verbenatin) relative to the weight of the dry extract.

[0031] "Hydrophilic solvents" refers to solvents selected from, in particular, water, subcritical water, water-miscible alcohols such as ethanol, C3 to C5 glycols, glycerol, acetone, and mixtures thereof.

[0032] For the purposes of this invention, "medium polar solvent" refers to a solvent selected particularly from C1 to C5 alcohols, glycols such as propylene glycol, butylene glycol, butanediol, or pentanediol, glycerol, acetone, alkyl esters such as ethyl acetate, isopropyl acetate, water-miscible solvents (e.g., water-alcohol mixtures or acetone / water mixtures), and mixtures of supercritical CO2 and ethanol. This group also includes alternative water-soluble growth promoters (water-soluble amphiphilic molecules that, at sufficient concentrations, can extract the medium polar compound as described in the characterization of the extract) or ionic liquids such as natural eutectic solvents (NaDES) formed from a mixture of hydrogen bond donor and hydrogen bond acceptor compounds in a defined proportion.

[0033] "Nonpolar solvents" refer to solvents selected from heptane, hexane, 2-methyltetrahydrofuran (MTHF), limonene, halogenated hydrocarbons (chloroform, dichloromethane), and supercritical CO2.

[0034] "Hair and body hair" refers to scalp hair, body hair, eyebrows, eyelashes and / or animal fur, with scalp hair being preferred.

[0035] "Skin appendages" refers to hair, body hair, eyebrows, eyelashes and / or nails, with hair being preferred.

[0036] "Hair loss" refers to the total or partial loss of hair, for example, due to reduced hair growth and / or accelerated hair loss. This term includes, but is not limited to, androgenetic alopecia, postmenopausal alopecia, reactive / emotional alopecia, cicatricial alopecia, alopecia areata, and congenital alopecia. The consequence of hair loss is temporary or permanent, partial or total hair loss. This discussion focuses primarily on reactive / emotional alopecia, and more specifically on telogen effluvium. Preferably, telogen effluvium is selected from acute and chronic telogen effluvium. In one embodiment, telogen effluvium is chronic telogen effluvium. In a preferred embodiment, telogen effluvium is acute telogen effluvium.

[0037] The term "treatment" of hair loss refers to stopping, reducing, and / or alleviating hair loss. Therefore, "treatment" of hair loss encompasses various stages of limiting hair loss and / or promoting hair growth, increasing hair follicle density, and / or regulating the hair follicle cycle. Specifically and preferably, it involves promoting and stimulating hair regeneration.

[0038] The term "preventing" hair loss refers to reducing the risk of hair loss or slowing the progression of hair loss in mammals (preferably men or women) that are prone to hair loss.

[0039] The term "limitation" refers to slowing down, reducing, lowering, and / or stopping.

[0040] The term "promote" means to increase, enlarge, benefit, amplify, and / or accelerate.

[0041] In this invention, "cosmetic or dermatologically acceptable" means a substance that is useful in the preparation of cosmetic or dermatological compositions, is generally safe and non-toxic, and is not biologically or otherwise undesirable, and is acceptable for cosmetic or dermatological use, particularly by topical application to hair and / or scalp.

[0042] "Topical application" refers to application to the skin, especially the scalp, mucous membranes and / or hair appendages, particularly the hair and / or scalp.

[0043] Explanation of the implementation plan Verbena extract This invention relates to a verbena extract for the use of preventing and / or treating hair loss.

[0044] In the context of this invention, verbena extract is obtained from one or more parts of the verbena plant, said parts being selected from the above-ground parts, such as stems, leaves, flowers, and fruits.

[0045] In one particular embodiment of the invention, the extract is obtained from a cell culture of verbena.

[0046] According to a particular embodiment of the invention, the extract is a hydroethanol extract, particularly a hydroethanol extract.

[0047] According to a preferred embodiment, the extract according to the invention can be obtained by the method described below.

[0048] Verbena plants or parts thereof can be fresh or dried, whole, cut or crushed, and then the extraction process is carried out.

[0049] The method for preparing the extract according to the invention includes the step of extracting whole or part of the verbena plant with a hydrophilic to nonpolar solvent, preferably a moderately polar solvent.

[0050] In one embodiment of the present invention, the extraction solvent may be selected from ethyl acetate, isopropyl acetate, a mixture of supercritical CO2 and ethanol, C3 to C5 diols, C1 to C5 alcohols, alcohol / water mixtures, water-soluble growth promoters, NaDES, or mixtures thereof.

[0051] Advantageously, it is an ethanol / water mixture or a water-soluble growth promoter solution.

[0052] More advantageously, it is an ethanol / water mixture.

[0053] Even more advantageously, the ethanol / water mixture is characterized by an ethanol / water ratio of 9:1 to 1:9 (v / v).

[0054] Even more advantageously, a moderately polar solvent is an ethanol / water mixture in a ratio of 7:3 (v / v).

[0055] Extraction can be carried out under stirring, in a dynamic recirculation system, or statically, under reflux, at room temperature, or at a temperature between room temperature and reflux. It can be assisted by ultrasound, microwave, flash expansion, or extrusion, with a plant weight / solvent volume ratio of 1 / 3 to 1 / 30, for a duration of 1 minute to 48 hours. Extraction can be repeated 2 to 3 times. Advantageously, verbena extraction is carried out with ultrasound assistance, with a plant weight / solvent volume ratio of 1 / 5 to 1 / 30, for a duration of 5 to 120 minutes.

[0056] The residue can then be separated from the extract by centrifugation or filtration to obtain a clear liquid phase free of particles. The liquid phase representing the extract can then be concentrated to varying degrees by evaporating the extraction solvent to obtain a dry extract.

[0057] A carrier may be added during the concentration step to obtain an extract containing 1 to 75% by weight of the dry extract. The carrier may be maltodextrin, lactose, silica, gum arabic or acacia gum, glycerin, glycol, vegetable oil, water-soluble growth promoter, water / solvent or water / surfactant mixture, or any other cosmetically acceptable carrier for dissolving the extract, preferably a bio-based carrier, such as bio-based glycols (1,2-pentanediol; 1,3-butanediol; 1,3-propanediol…), esterified fatty acids, and water-soluble growth promoters, such as alkyl glycosides (Sepiclear, Apyclean, APXC4…). Advantageously, the verbena extract is standardized in a maltodextrin matrix or a glycerin / water mixture.

[0058] Verbena extract can be decolorized to remove chlorophyll, for example, using activated charcoal or by concentration / precipitation / filtration. Advantageously, verbena extract is decolorized by concentration / precipitation / filtration.

[0059] Verbena extract can be characterized by the presence of at least one phytochemical family of glycosylated iridoids, glycosylated phenylethyl glycosides, or flavonoids.

[0060] Verbena extract may have a content of 0.1% to 60%, particularly 1% to 30%, more particularly 2% to 20%, or even more particularly about 10% of glycosylated iridoid compounds relative to the weight of the dry extract.

[0061] Verbena extract may have a content of 0.01 to 60%, particularly 1 to 40%, or even more particularly about 10% of glycosylated phenylethanol glycosides relative to the weight of the dry extract.

[0062] Verbena extract may have a flavonoid content of 0.01 to 50%, particularly 0.01 to 20%, or even more particularly about 5%, relative to the weight of the dry extract.

[0063] Verbena extract may have a content of 1 to 15% by weight relative to the dry extract, particularly 5 to 15% verbenacin.

[0064] Verbena extract, as described above, can prevent or treat hair loss, particularly selected from androgenetic alopecia, reactive alopecia, postmenopausal alopecia, and alopecia areata.

[0065] Preferably, verbena extract can be used to treat or prevent androgenetic alopecia.

[0066] Even more preferably, the verbena extract according to the invention can prevent or treat reactive / emotional alopecia, preferably telogen effluvium. According to one embodiment, the verbena extract according to the invention can prevent or treat acute telogen effluvium. According to one embodiment, the verbena extract according to the invention can prevent or treat chronic telogen effluvium.

[0067] Preferably, the extract according to the invention is used as an anti-hair loss active ingredient. Advantageously, the extract according to the invention is used to promote hair regeneration.

[0068] In particular, the extract according to the invention stimulates hair regeneration. More specifically, hair regeneration is achieved by stimulating VGEF.

[0069] The extract helps limit hair loss and / or promote hair growth and / or increase hair follicle density and / or achieve denser hair coverage and / or promote hair follicle regeneration. Preferably, the prevention and / or treatment of hair loss is achieved through one or more of the following mechanisms: • It prolongs the lifespan of hair by inhibiting the activation of the JAK / STAT pathway in the outer root sheath cells. • It stimulates blood vessel growth to promote hair regeneration by producing VEGF. • It limits hair loss and stimulates hair growth by inhibiting apoptosis of outer root sheath cells induced by prostaglandin D2 metabolites.

[0070] Dermatological or cosmetic compositions The present invention also relates to a dermatological or cosmetic composition for the prevention and / or treatment of hair loss, said dermatological or cosmetic composition comprising at least one verbena extract and at least one dermatologically or cosmetically acceptable excipient.

[0071] Hair loss may be specifically selected from androgenetic alopecia, reactive alopecia, postmenopausal alopecia, and alopecia areata; preferably, it is androgenetic alopecia. According to a particular embodiment, it is reactive / emotional alopecia, preferably telogen effluvium.

[0072] According to one embodiment, the composition according to the invention is capable of preventing or treating acute telogen effluvium. According to another embodiment, the composition according to the invention is capable of preventing or treating chronic telogen effluvium.

[0073] The composition helps to limit hair loss and / or promote hair growth and / or increase hair follicle density and / or achieve denser hair coverage and / or promote hair follicle regeneration.

[0074] Advantageously, extracts included in dermatological or cosmetic compositions are as described above.

[0075] Advantageously, the cosmetic or dermatological composition comprises an extract obtained by hydro-alcohol extraction, i.e., a hydro-alcohol extract of verbena, particularly an ethanol extract, more specifically an ethanol / water mixture, and even more advantageously, the ethanol / water mixture being characterized by an ethanol / water ratio of 9:1 to 1:9 (v / v). Advantageously, the ethanol / water mixture is an ethanol / water mixture with a ratio of 7:3 (v / v).

[0076] Cosmetic or dermatological compositions may contain 0.001 to 5%, preferably 0.002 to 2%, preferably 0.005 to 1%, or even more preferably 0.01 to 0.5% of verbena dry extract relative to the total weight of the composition.

[0077] The present invention preferably relates to cosmetic or dermatological compositions suitable for topical application, particularly for topical application to the scalp and / or hair.

[0078] Therefore, the cosmetic or dermatological compositions according to the invention can be in forms commonly known for topical application, i.e., in particular lotions, shampoos, balms, foams, gels, dispersions, emulsions, sprays, serums, masks, or creams, having excipients that particularly allow penetration to improve the properties and accessibility of the active ingredients.

[0079] Advantageously, the compositions according to the invention can be in forms commonly known for topical application to hair and scalp, i.e., in particular shampoos, conditioners, hair creams, hair liquids, hair masks, or dry shampoo sprays.

[0080] Therefore, it is possible to distinguish between rinseable formulations and formulations that do not require rinsing.

[0081] Preferably, the composition according to the invention has a light texture, which also allows for optimal penetration without making hair and / or body hair, or scalp oily.

[0082] The compositions according to the invention (e.g., for topical application) typically contain a physiologically acceptable medium other than the extracts described above, which is usually water-based or solvent-based, such as an alcohol, ether, or glycol. It may also contain surfactants, complexing agents, preservatives, stabilizers, emulsifiers, thickeners, gelling agents, humectants, emollients, trace elements, essential oils, fragrances, colorants, moisturizers, or thermal spring water, etc.

[0083] From the first application (e.g., topical or oral) of the composition according to the invention, hair will regain strength and vitality and will promote regeneration.

[0084] Therefore, the extracts or compositions described above can be used to promote hair regeneration.

[0085] According to one embodiment of the invention, the verbena extract as described above, or a cosmetic or dermatological composition comprising at least one verbena extract and at least one cosmetically or dermatologically acceptable excipient as described above, is intended for topical application.

[0086] The present invention also relates to a cosmetic method comprising applying a composition containing the above-mentioned verbena extract to hair.

[0087] This invention relates to the use of verbena extract as defined above, or dermatological compositions comprising at least one verbena extract and at least one dermatologically acceptable excipient, for the prevention and / or treatment of hair loss.

[0088] The present invention also relates to the use of verbena extract as defined above or a dermatological composition comprising at least one verbena extract and at least one dermatologically acceptable excipient in the preparation of a medicament or dermatological composition intended to promote hair growth by preventing hair loss.

[0089] This invention also relates to the use of verbena extract as defined above, or a dermatological composition comprising at least one verbena extract and at least one dermatologically acceptable excipient, for promoting hair regeneration. Advantageously, this invention relates to the use of extracts or compositions according to the invention for the prevention or treatment of reactive / emotional alopecia, more particularly telogen effluvium, preferably by promoting hair regeneration. According to a particular embodiment, this invention relates to the use of extracts or compositions according to the invention for the prevention or treatment of chronic telogen effluvium. In a preferred embodiment, this invention relates to the use of extracts or compositions according to the invention for the prevention or treatment of acute telogen effluvium.

[0090] The present invention also relates to a method for promoting hair growth, particularly for preventing and / or treating hair loss, especially reactive / emotional hair loss, and more particularly telogen effluvium, comprising administering to a patient in need an effective amount of verbena extract as defined above or a cosmetic or dermatological composition comprising at least one verbena extract and at least one cosmetically or dermatologically acceptable excipient.

[0091] This invention also relates to the use of verbena extract as described above or cosmetic or dermatological compositions as described above for hair and / or scalp care, and / or for promoting hair growth, and / or increasing hair follicle density, and / or achieving denser hair coverage, and / or promoting hair follicle regeneration, and / or for cosmetic use in combating hair loss, which may be selected from androgenetic alopecia, postmenopausal alopecia, more particularly reactive / emotional alopecia, and more particularly telogen effluvium. The cosmetic use according to the invention particularly relates to promoting hair growth to combat reactive / emotional alopecia, and more particularly telogen effluvium.

[0092] This invention also relates to a non-therapeutic cosmetic method for hair and / or scalp care, and / or for limiting hair loss and / or promoting hair growth, and / or increasing hair follicle density and / or achieving denser hair coverage and / or promoting hair follicle regeneration and / or combating hair loss, wherein the hair loss may be selected from androgenetic alopecia, postmenopausal alopecia, particularly reactive / emotional alopecia, and more particularly telogen effluvium, comprising applying verbena extract as described above or cosmetic compositions as described above to the hair and / or scalp. In a particular embodiment of the method according to the invention, the hair loss is chronic telogen effluvium. In a particularly preferred embodiment of the method according to the invention, the hair loss is acute telogen effluvium. Therefore, the method according to the invention is particularly intended to promote hair growth to combat reactive / emotional alopecia, and more particularly telogen effluvium. The present invention also relates to a non-therapeutic cosmetic method for preventing and / or treating hair loss, particularly for promoting hair growth, such as androgenetic alopecia, postmenopausal alopecia, especially reactive / emotional alopecia, and even more particularly telogen effluvium, comprising the use of verbena extract as defined above or a cosmetic or dermatological composition comprising at least one verbena extract and at least one cosmetically or dermatologically acceptable excipient.

[0093] Example The following examples illustrate the invention but do not limit its scope. Examples 1 to 5 illustrate different extraction types of the above-ground parts of verbena.

[0094] Example 1: Ultrasonic-assisted extraction with 70% ethanol 34 g of pulverized, dried verbena upper parts were contacted with 340 mL of 70% ethanol and then extracted with ultrasound (1000 W, 20 kHz) for 5 min with stirring. The extract was then filtered under vacuum using a Buchner funnel. The extract was subsequently decolorized by pre-concentration, cooled at 4°C for 12 h, and filtered again. The solvent was then evaporated to give 3.54 g of powder, with a yield of 10.4%. The resulting dry extract contained 7.3% verbenaside (a glycosylated iridoid compound specific to verbena) by weight.

[0095] Example 2: Microwave-assisted extraction with 90% ethanol 1.5 g of pulverized, dried verbena upper part was contacted with 15 mL of 90% (v / v) ethanol and then extracted using microwave (850 W, up to 50°C) for 30 min. The extract was then filtered under vacuum using a Buchner funnel. The solvent was evaporated to obtain a dry powder extract with a yield of 6.6% by mass and a verbenacin content of 9.4%.

[0096] Example 3: Extraction with 50% ethanol under reflux Five grams of pulverized, dried verbena upper parts were contacted with 50 mL of 50% (v / v) ethanol and then extracted by reflux with stirring for 60 minutes. The extract was then filtered under vacuum using a Buchner funnel. The solvent was then evaporated to give 0.9 g of powder, with a yield of 18%. The resulting dry extract contained 5.3% verbenacin by weight.

[0097] Example 4: Reflux extraction with ethyl acetate Five grams of pulverized, dried verbena upper parts were contacted with 50 mL of ethyl acetate and then extracted by reflux with stirring for 60 minutes. The extract was then filtered under vacuum using a Buchner funnel. The solvent was then evaporated to give 0.25 g of powder, with a yield of 5%. The resulting dry extract contained 8.5% verbenacin by weight.

[0098] Example 5: Ultrasonic-assisted extraction of 2-methyltetrahydrofuran (MTHF) Five grams of pulverized, dried verbena upper parts were mixed with 50 mL of MTHF in a beaker and then extracted with ultrasound (1000 W, 20 kHz) for 1 minute with stirring. The extract was then filtered under vacuum using a Buchner funnel. The solvent was then evaporated to give 0.17 g of powder, with a mass yield of 10.4%. The resulting dry extract contained 3.4% verbenacin by weight.

[0099] Example 6: Evaluation of the inhibitory effect of verbena extract on 15-dPGJ2 The hair growth cycle is a highly regulated biological mechanism during which the different compartments of the hair follicle (dermal papilla, resident stem cells of the hair follicle, and keratinocytes of the hair follicle matrix) maintain close relationships through various molecular exchanges. Recently, prostaglandin D2 (PGD2) has been overexpressed in bald areas compared to hairy areas on the scalp of men with androgenetic alopecia (Garza, Luis et al., Science Translational Medicine 2012, Vol. 4, No. 126, p126ra34). Consistent with these observations, DP2 (a PGD2 receptor) is overexpressed in the outer root sheath of keratinocytes in bald areas.

[0100] The metabolite 15-deoxy-12,14-prostaglandin (15d-PGJ2) is a non-enzymatic compound derived from PGD2 that has been identified as a pro-apoptotic factor, i.e., it induces apoptosis in outer root sheath cells and induces them to enter the regression phase (Joo, Hyun Woo et al., Naunyn-Schmiedeberg's Archives of Pharmacology 2016, Vol. 389, No. 7, p809613).

[0101] This study demonstrates that the metabolite 15d-PGJ2 induces apoptosis, which in turn leads to hair loss.

[0102] The purpose of this embodiment is to evaluate whether verbena extract can inhibit the apoptosis pathway induced by metabolite 15d-PGJ2.

[0103] Experiments were performed on keratinocytes from the outer root sheath of hair follicles from three human donors. Cells were seeded in 96-well plates and cultured for 24 hours with the necessary supplements. The medium was then changed, and the cells were treated for 24 hours with the product to be tested (verbena officinalis extract diluted in DMSO at concentrations of 3, 10, or 30 µg / ml) and 5 µM of metabolite 15-dPGJ2. After 24 hours of incubation, caspase 3 / 7 activity was measured on the cell layers by luminescence (Promega G6411) according to the manufacturer's instructions. All experimental conditions were performed in n=3 donor and n=3 technique replicates.

[0104] The cell model was validated and cytotoxicity was verified using verbena extract diluted in DMSO. No difference was found between treatment with 0.1% v / v DMSO and untreated cells. Therefore, it can be concluded that the tested concentration of DMSO does not alter cell viability and will be used as a control in subsequent experiments.

[0105] Next, after treatment with the metabolite 15d-PGJ2 in combination with a known caspase inhibitor such as Z-VAD-FMK (Sigma), caspase 3 / 7 activity was measured.

[0106] [Table 1] SEM: Standard Error of Mean The metabolite 15d-PGJ2 induced apoptosis in outer root sheath cells. The percentage of apoptosis significantly decreased upon the addition of compound Z-VAD-FMK, and was similar to the control value. The addition of compound Z-VAD-FMK showed a significant reduction in 15d-PGJ2-induced apoptosis, which reflects a reduction in hair loss.

[0107] [Table 2] The verbena extract obtained according to Example 1 was tested at three different concentrations: 3, 10, and 30 µg / ml. A reduction in apoptosis was observed in all three verbena extracts in a concentration-dependent manner.

[0108] Verbena officinalis extract induced concentration-dependent inhibition of 15d-PGJ2-induced apoptosis in outer root sheath cells (40% inhibition at 5 µg / ml and 76% inhibition at 30 µg / ml compared to the untreated control).

[0109] Therefore, verbena extract acts on prostaglandin D2 and helps limit hair loss by inhibiting apoptosis of outer root sheath cells, which helps promote hair growth.

[0110] Example 7: Evaluation of the inhibitory effect of verbena extract on the VEGF pathway It is known that VEGF is produced by dermal papillary cells of hair follicles (Lachgar (1996) J Invest Dermatol, 106(1): 17-23). ​​It is also known (Kubanov et al., Biomedical & Pharmacology Journal Vol. 10(1),191-198 (2017)) that emotional / reactive alopecia, or telogen effluvium, is characterized by diffuse hair loss during the telogen phase, often triggered by factors such as psychological stress. This study shows that all the growth factors studied (VEGF, KGF, EGF, TGF-β1) contribute to the development of telogen effluvium, with VEGF having the greatest impact. Therefore, a significant decrease in VEGF expression was observed in the scalp. VEGF plays a crucial role in angiogenesis and vascularization, thereby stimulating hair growth. Therefore, decreased VEGF expression in hair follicles is a key factor in the development of this form of hair loss. Conversely, stimulation of it promotes hair regeneration.

[0111] VEGF is a growth factor for endothelial cells; it promotes the supply of oxygen and nutrients to hair follicles. The expression level of this protein varies throughout the hair growth cycle: it is mainly produced in the early stages of hair growth (anagen phase) and decreases as the hair follicle enters the mid-stage (catagen phase). VEGF promotes the formation of new blood vessels, thus stimulating endothelial cells. Therefore, when induced, this growth factor stimulates angiogenesis, thereby promoting hair regeneration.

[0112] The purpose of this embodiment is to evaluate whether verbena extract can stimulate the production of VEGF factors, thereby restoring and stimulating hair regeneration and growth.

[0113] The experiment was conducted on human follicular dermal papillary cells (HFDPC) from three different donors.

[0114] Cells were seeded in 96-well plates and cultured for 24 hours in HFDPC medium (Promocell, C-26500) supplemented with an appropriate supplement (Promocell, C-39625). The medium was then replaced with test medium (HFDPC medium from Promocell without supplementation). The test medium may or may not contain the (control) compound being tested or a reference (manganese salt of pyrrolidone carboxylic acid (PCA)). Cells were incubated for 24 hours.

[0115] The culture supernatant was then collected for VEGF assay. All experimental conditions were performed at n=3.

[0116] [Table 3] Significant induction of VEGF release was observed in the presence of PCA manganese salt (which served as an internal reference) (+118% compared to the untreated control). This result was expected and validated the assay.

[0117] The verbena extract obtained according to Example 1 induced an increase in VEGF release from dermal papilla cells in a concentration-dependent manner (+23% at 10 µg / ml and +53% at 30 µg / ml compared to the untreated control).

[0118] Therefore, verbena extract leads to the production of VEGF factors and promotes hair regeneration by stimulating blood vessel growth.

[0119] Example 8: Evaluation of Verbena officinalis extract on the JAK-STAT pathway The aim of this embodiment was to confirm the inhibitory activity of Verbena officinalis extract on the JAK-STAT signaling pathway in a cell model, specifically, follicular keratinocytes (ORS cells) of the outer root sheath of hair follicles. In this model, interleukin IL-6 is used to activate the JAK1 / 2-STAT3 pathway by activating IL-6 and GP130 receptors, both of which are expressed in the root sheath of hair follicles. IL-6 is a cytokine that inhibits the hair growth cycle; its overexpression in a transgenic mouse model leads to delayed hair growth. IL-6 has also been reported to delay hair follicle growth in humans.

[0120] The JAK1 / 2-STAT3 pathway, activated by IL-6 and GP130 receptors, is expressed in the root sheath of hair follicles. This activation plays a crucial role in regulating hair growth. Overactivation of this pathway can contribute to hair loss by inducing inflammation and an autoimmune response against the hair follicle. JAK inhibitors can help treat this condition by reducing the activation of the JAK-STAT pathway, thereby modulating inflammation and promoting hair regrowth.

[0121] The study was conducted on follicular keratinocytes derived from the outer root sheath of hair follicles (ORS model). Keratinocytes were cultured in 96-well plates in a suitable medium (Cellntec CnT-PR). After 48 hours, the cells were washed with alkaline phosphate-buffered saline (PBS), and the medium was replaced with KSFM (standard maintenance medium for primary human keratinocytes). After 4 hours, the cells were treated for one hour with the test compound (verbena officinalis extract diluted in DMSO at 10, 30, and 100 µg / ml) and the reference compound (tofacitinib diluted in DMSO at 5 µM). The verbena officinalis extract used in the study was the extract according to Example 1 of the present invention. Then, IL-6 stimulation (Stim IL6) was performed at 100 ng / ml for 15 minutes.

[0122] All experimental conditions were performed with n=3 donors and n=3 techniques in a repeat.

[0123] The activation level of the JAK / STAT signaling pathway was estimated by assessing the phosphorylation level of the STAT3 protein. Statistical analysis was performed using parametric tests after validating normality and variance equivalence; otherwise, nonparametric tests were used.

[0124] [Table 4] Treatment of keratinocytes with IL-6 resulted in a significant increase in STAT3 phosphorylation levels; this increase was statistically significant (P < 0.0001). The reference compound tofacitinib, tested at 5 µM, significantly reduced STAT3 phosphorylation levels by 97.5% (p < 0.0001). These expected results validated the assay.

[0125] The verbena extract obtained according to Example 1 significantly and very markedly inhibited IL-6-induced STAT3 phosphorylation levels at all three tested concentrations. Therefore, it exerted its effect in a concentration-dependent manner starting from 10 µg / ml (p = 0.0001).

[0126] The inventors therefore emphasized the benefits of verbena extract in hair loss by extending the hair life cycle through significant inhibition of JAK / STAT pathway activation induced by IL6 in the outer root sheath cells.

[0127] In summary, the experimental evidence demonstrating the effects of the extracts according to the invention on vascular endothelial growth factor (VEGF) activation and JAK-STAT pathway inhibition offers promising prospects for the treatment of hair loss, particularly emotional / reactive alopecia or telogen effluvium. VEGF activation promotes angiogenesis and hair follicle nutrition, thereby stimulating hair growth and regeneration. Simultaneously, inhibition of the JAK-STAT pathway reduces inflammation and autoimmune responses, protecting hair follicles from damage. These combined effects not only prevent hair loss associated with stress and emotional responses but, most importantly, positively stimulate hair regeneration.

[0128] Therefore, the extracts according to the invention have been shown to be a natural and effective solution for promoting hair regeneration, hair growth and treating forms of hair loss associated with stress and emotional responses, such as reactive / emotional alopecia or telogen effluvium.

[0129] Implementation scheme of the invention 1. An aqueous ethanol extract of Verbena officinalis, used as an anti-hair loss active ingredient in dermatological or cosmetic compositions to promote hair regeneration as part of the prevention and / or treatment of hair loss.

[0130] 2. The extract according to the use described in Embodiment 1, wherein the prevention and / or treatment of hair loss is achieved through one or more of the following mechanisms: • It prolongs the lifespan of hair by inhibiting the activation of the JAK / STAT pathway in the outer root sheath cells. • It stimulates blood vessel growth to promote hair regeneration by producing VEGF. • It limits hair loss and stimulates hair growth by inhibiting apoptosis of outer root sheath cells induced by prostaglandin D2 metabolites.

[0131] 3. An extract for use according to embodiment 1 or 2, wherein the hair loss is selected from androgenetic alopecia, reactive / emotional alopecia (especially telogen effluvium), postmenopausal alopecia and alopecia areata.

[0132] 4. The extract according to the use described in embodiment 3, characterized in that the hair loss is reactive / emotional hair loss.

[0133] 5. The extract for use according to embodiment 4, characterized in that reactive / emotional alopecia is telogen effluvium.

[0134] 6. An extract for use according to any one of embodiments 1 to 6, for obtaining thicker hair coverage and / or promoting hair follicle regeneration.

[0135] 7. The extract according to any one of embodiments 1 to 7, characterized in that the extract is in a form suitable for topical application to the scalp.

[0136] 8. The extract for use according to any one of embodiments 1 to 8, characterized in that the verbena extract contains 1% to 15% verbenin.

[0137] 9. A dermatological or cosmetic composition for use in promoting hair regrowth in the prevention and / or treatment of hair loss, said dermatological or cosmetic composition comprising at least one aqueous ethanol extract of verbena as an anti-hair loss active ingredient and at least one dermatologically or cosmetically acceptable excipient, characterized in that the hair loss is reactive / emotional hair loss, more particularly telogen effluvium.

[0138] 10. The composition for use according to embodiment 9, wherein the prevention and / or treatment of hair loss is achieved through one or more of the following mechanisms: • It prolongs the lifespan of hair by inhibiting the activation of the JAK / STAT pathway in the outer root sheath cells. • It stimulates blood vessel growth to promote hair regeneration by producing VEGF. • It limits hair loss and stimulates hair growth by inhibiting apoptosis of outer root sheath cells induced by prostaglandin D2 metabolites.

[0139] 11. A dermatological or cosmetic composition for use according to any one of embodiments 10 or 11, characterized in that it comprises 0.001% to 10% by weight of verbena dry extract relative to the total weight of the composition, the dry extract comprising 1% to 15% verbena glycoside.

[0140] 12. A dermatological or cosmetic composition for use according to any one of embodiments 9 to 11, characterized in that the composition is in a form suitable for topical application to the scalp.

[0141] 13. A dermatological or cosmetic composition for use according to any one of embodiments 9 to 12, characterized in that the composition is capable of limiting hair loss and / or promoting hair growth and / or increasing hair follicle density and / or achieving denser hair coverage and / or promoting hair follicle regeneration.

[0142] According to a particularly preferred embodiment, the present invention relates to an aqueous ethanol extract of Verbena officinalis, which is used as an anti-hair loss active ingredient in dermatological or cosmetic compositions to promote hair regeneration as part of the prevention and / or treatment of hair loss, characterized in that the hair loss is reactive / emotional hair loss, and more particularly telogen effluvium.

[0143] More specifically, the present invention relates to an aqueous ethanol extract of Verbena officinalis, the use of which as an anti-hair loss active ingredient in dermatological or cosmetic compositions to promote hair regeneration in the prevention and / or treatment of hair loss, characterized in that the hair loss is reactive / emotional hair loss, more particularly telogen effluvium, wherein the prevention and / or treatment of hair loss is achieved through one or more of the following mechanisms: • Extending the lifespan of hair by inhibiting the activation of the JAK / STAT pathway in outer root sheath cells. • It stimulates blood vessel growth to promote hair regeneration by producing VEGF. • It limits hair loss and stimulates hair growth by inhibiting apoptosis of outer root sheath cells induced by prostaglandin D2 metabolites.

Claims

1. A type of verbena ( Verbena officinalis Extracts, used for the prevention and / or treatment of reactive / emotional alopecia.

2. The extract according to claim 1, characterized in that... Reactive / emotional alopecia is telogen effluvium.

3. The extract according to claim 1 or 2, characterized in that... The extract is used as an anti-hair loss active ingredient to promote hair regeneration.

4. The extract according to any one of claims 1 to 3, for limiting hair loss and / or promoting hair growth and / or increasing hair follicle density and / or obtaining denser hair coverage and / or promoting hair follicle regeneration.

5. The extract according to any one of claims 1 to 4, characterized in that... The verbena extract is an aqueous ethanol extract of verbena.

6. The extract according to any one of claims 1 to 5, characterized in that... The verbena extract has a verbena content of 1% to 15% by weight relative to the dry extract.

7. The extract according to any one of claims 1 to 6, characterized in that... The extract is in a form suitable for topical application.

8. A cosmetic or dermatological composition for the prevention and / or treatment of hair loss, comprising verbena extract and at least one dermatologically or cosmetically acceptable excipient, characterized in that... The hair loss described is reactive / emotional hair loss.

9. The cosmetic or dermatological composition for use according to claim 8, characterized in that... Reactive / emotional alopecia is telogen effluvium.

10. A cosmetic or dermatological composition for use according to claim 8 or 9, for promoting hair regeneration.

11. A cosmetic or dermatological composition for use according to any one of claims 8 to 10, characterized in that... The extract is used as an active ingredient in anti-hair loss treatment.

12. A cosmetic or dermatological composition for use according to any one of claims 8 to 11, characterized in that... The verbena extract is an aqueous ethanol extract of verbena.

13. A cosmetic or dermatological composition for use according to any one of claims 8 to 12, characterized in that... It contains 0.001% to 5% by weight of verbena dry extract relative to the total weight of the composition.

14. A cosmetic or dermatological composition for use according to any one of claims 8 to 13, characterized in that... The composition is in a form suitable for topical application.

15. A cosmetic or dermatological composition for use according to any one of claims 8 to 14, characterized in that... The composition can limit hair loss and / or promote hair growth and / or increase hair follicle density and / or achieve denser hair coverage and / or promote hair follicle regeneration.

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