Composition comprising an extract of rhodiola rosea, tryptophan and vitamin k for preventing or reducing hair loss due to stress

Abstract

The invention relates to a cosmetic and / or pharmaceutical composition for preventing or reducing hair loss of a mammal due to stress and for use in the treatment of a hair growth disease or hair growth disorder exacerbated by stress.

Description

[0001] P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0002] TITLE:

[0003] COMPOSITION FOR PREVENTING OR REDUCING HAIR LOSS DUE TO

[0004] STRESS

[0005] ********************

[0006] FIELD OF THE INVENTION

[0007] The present invention relates to a composition for preventing or reducing hair loss due to stress.

[0008] In the present invention when the term “hair” is used, it is intended to comprise also skin appendages, i.e. beard, eyelashes, eyebrows and nails.

[0009] Specifically, the invention relates to a composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K.

[0010] The composition can be used in cosmetics and can be used in the treatment of hair growth disorders or hair growth diseases that are exacerbated by stress.

[0011] STATE OF THE ART

[0012] Stress is broadly defined as the body’s reaction to external harmful stimuli that disrupts its internal equilibrium. This response involves various psychophysical and behavioral changes designed to enable defense mechanisms. Stress can be categorized in two key ways: homeostasis, which maintains vital physiological parameters (e.g., pH, temperature, blood oxygen levels) within specific ranges essential for survival, and allostasis, which reflects physiological adjustments (like heart rate and blood pressure) in response to stressors. Initially, these adaptations can be beneficial, enhancing performance. However, if stress persists or becomes overwhelming, it leads to strain on the body’s adaptive systems, resulting in functional disturbances and eventually serious health issues.

[0013] The relationship between stress and hair growth is a complex interplay of physiological mechanisms, primarily mediated by hormonal changes, particularly the elevation of cortisol levels (Thom E. (2016). Stress and the Hair Growth Cycle: Cortisol-Induced Hair Growth Disruption. Journal of drugs in dermatology: JDD, 15(8), 1001-1004). Stress, whether acute or chronic, has been shown to significantly affect the hair growth cycle, leading to conditions such as telogen effluvium, where hair follicles prematurely enter the resting phase, resulting in increased hair shedding (Thom E. (2016). Stress and the Hair Growth Cycle: P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0014] Cortisol-Induced Hair Growth Disruption. Journal of drugs in dermatology: JDD, 15(8), 1001-100). The mechanisms underlying these effects are multifaceted, involving neuroendocrine pathways, immune responses, and direct cellular interactions within hair follicles.

[0015] Cortisol, a glucocorticoid hormone produced by the adrenal glands, plays a pivotal role in the body’s response to stress. Under stress, cortisol levels rise, which can inhibit hair growth by disrupting the normal hair follicle cycle (Thom E. (2016). Stress and the Hair Growth Cycle: Cortisol-Induced Hair Growth Disruption. Journal of drugs in dermatology: JDD, 15(8), 1001-1004). Research has demonstrated that acute stress can lead to a two-fold increase in cortisol levels, which correlates with inhibited hair growth in animal models. This elevation in cortisol is not merely a transient response; chronic stress conditions can lead to sustained high levels of cortisol, further exacerbating hair loss.

[0016] The hair growth cycle consists of three distinct phases: anagen (growth), catagen (regression), and telogen (resting). Stress-induced hormonal changes can accelerate the transition from anagen to telogen, leading to increased hair shedding and reduced overall hair density. The mechanisms by which cortisol affects hair follicles involve several pathways. Cortisol can influence the dermal papilla cells, which are critical for hair follicle growth and cycling. Studies have shown that stress can lead to changes in the morphology and function of these cells, potentially impairing their ability to support hair growth. Acute stress-induced changes in follicular dermal papilla cells and mobilization of mast cells: implications for hair growth. Additionally, stress can mobilize mast cells in the skin, which release inflammatory mediators that can further disrupt hair follicle function and promote hair loss.

[0017] The role of neuropeptides and cytokines in this process cannot be overlooked, as they are also implicated in the modulation of hair follicle activity during stress. Moreover, the incorporation of cortisol into hair shafts serves as a biomarker for chronic stress, providing insights into the long-term effects of stress on hair growth. Hair cortisol concentration (HCC) reflects the cumulative exposure to cortisol over time, with each centimeter of hair representing approximately one month of growth. This method allows researchers to assess the impact of stress on hair growth P024192WO-01 Notarbartolo & Gervasi S.p.A. retrospectively, revealing patterns that correlate with periods of heightened stress. The relationship between stress and hair growth is further complicated by individual differences in hair growth rates and hair follicle cycling. Factors such as genetics, age, and overall health can influence how stress affects hair growth. For instance, hair growth rates can vary significantly across different regions of the scalp, and the physiological response to stress may differ based on these variations. Additionally, the presence of other hormones, such as androgens, can interact with cortisol, influencing hair follicle behavior and potentially leading to conditions like androgenetic alopecia.

[0018] Furthermore, the psychological aspects of stress, including anxiety and depression, can also contribute to hair loss. The mind-body connection is well-documented, with psychological stressors leading to physiological changes that can impact hair growth. For example, studies have indicated that individuals with high levels of psychological stress may exhibit increased hair loss, further linking mental health to physical manifestations such as alopecia.

[0019] A recent study highlighted the role of corticosterone, the mouse equivalent of human cortisol, in regulating hair follicle stem cell (HFSC) behavior. This research indicated that systemic conditions, such as chronic stress, impact HFSC activity, an area that is not yet fully understood. The study specifically examined how the adrenal glands, responsible for producing stress hormones, affect hair growth. Mice that underwent adrenal gland removal displayed significantly shorter telogen phases compared to control animals, indicating heightened hair growth activity. This suggests that stress hormones play a critical role in quiescence and activation of HFSCs.

[0020] Further exploring the mechanisms, it was found that in the bulge niche of hair follicles, various regulatory molecules influence HFSC behavior. During the early stages of telogen, growth factors secreted by neighboring cells keep HFSCs inactive, whereas in late telogen, high levels of corticosterone can suppress key molecules like Gas6, inhibiting the transition to the anagen phase and thus prolonging hair follicle dormancy. This highlights a dual role of stress mediators in both targeting and being generated by cellular processes associated with hair growth.

[0021] Stress induces systemic (general) production of cortisol which, through the blood P024192WO-01 Notarbartolo & Gervasi S.p.A. stream, reaches the follicle, attacking it and causing it to fall.

[0022] Also at the local level (of the hair follicle), in response to stress, an increase in the production of cortisone is then converted into cortisol by the enzyme 11 beta-HSD1 : 11 [3-hydroxyasteroid dehydrogenase type 1 . Corticosterone, a hormone produced by the adrenal gland of rodents as a result of stress, is the equivalent of cortisol in humans.

[0023] This hormone (and cortisol in humans) regulates the quiescence of hair follicle stem cells (HFSC) and hair growth in mice. Under chronic stress, increased corticosterone levels prolong the HFSC quiescence and keep hair follicles in a prolonged resting phase anticipating fall and delaying new growth. Mechanically, corticosterone acts on the dermal papillae to suppress the expression of Gas6, a gene that encodes for a protein called "growth-stopping factor specific 6". Gas6 is a protein containing the gamma-carboxylglutamic acid (Gia) domain which is believed to be involved in stimulating cell proliferation.

[0024] The connection between psychological stress and hair loss has become increasingly relevant, particularly in light of recent events like the COVID-19 pandemic. Conditions such as telogen effluvium (TE) and alopecia areata (AA) are well-recognized reversible forms of hair loss that may stem from or be worsened by perceived stress. Various interactions between stress and hair loss can be identified. For instance, stress can directly trigger TE or act as a complicating factor in disorders with different primary causes. Additionally, stress can occur as a secondary response to existing hair loss, potentially creating a vicious cycle. About one-quarter of individuals recovering from COVID-19 report experiencing hair loss approximately six months after symptoms appear. This may stem from the systemic shock of the infection and the subsequent recovery process. Data during the pandemic revealed that 42.5% of participants with a history of alopecia areata experienced a relapse following COVID-19 infection, typically within two months post-infection. A systematic review conducted in 2022 further pointed out that while some studies found exacerbation of AA following SARS-CoV-2 infection, the variability among reported cases makes it challenging to establish definitive correlations.

[0025] The worsening of alopecia areata can be considered multifactorial. Physical stress P024192WO-01 Notarbartolo & Gervasi S.p.A. resulting from an infection like COVID-19, combined with the considerable psychosocial challenges stemming from a pandemic, may significantly contribute to the deterioration of underlying dermatological conditions. The intricate balance between stress, immune response, and hair follicle health reveals a complex interplay that necessitates further investigation to understand better the dynamics of hair loss in stressful contexts.

[0026] The inventors were aware that the effects of stress on hair growth are mediated through a complex network of hormonal, immune, and psychological factors. The elevation of cortisol during stress plays a central role in disrupting the hair growth cycle, leading to increased shedding and potential long-term hair loss.

[0027] Therefore the main object is managing stress in an effective way in order to preserve hair health and to mitigate hair loss conditions such as alopecia areata and other forms of alopecia, temporary or permanent hair loss conditions.

[0028] SUMMARY OF THE INVENTION

[0029] In general terms, the present invention provides new uses in the cosmetic and trichology fields of a combination of biologically active ingredients.

[0030] The inventors have found that an extract of Rhodiola Rosea, tryptophan and Vitamin K synergistically act by guaranteeing or increasing hair health by preventing or reducing the hair loss when the extract of Rhodiola Rosea and tryptophan are in specific amounts.

[0031] Therefore in a first aspect the invention relates to a composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K, wherein the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition.

[0032] In the present invention when the term “hair” is used, it is intended to comprise also skin appendages, i.e. beard, eyelashes, eyebrows and nails, of a mammal.

[0033] Therefore “hair”, includes hair and skin appendages such as, preferably, beard, eyelashes, eyebrows and nails of a mammal which is a human being, or an animal being, for example a dog or cat or another pet or companion animal.

[0034] According to the invention the mammal is preferably a human being.

[0035] Advantageously, a specific subject of the invention is hence a non-therapeutic, P024192WO-01 Notarbartolo & Gervasi S.p.A. cosmetic use of the composition of the invention in promoting hair growth in a mammal, in particular a human being, specifically when subjected to stress.

[0036] In accordance with certain specific aspects of the invention, the hair loss reduction activity of the composition described herein affects a population of individuals who are affected by a greater than physiological hair loss and which can cause thinning on the scalp.

[0037] The composition of the invention that finds application in the cosmetic field by preventing or reducing the hair loss, preferably due to stress, allows the achievement of a desired external aspect of hair and skin appendages, such as for example their volume, their shine and / or their compactness, their light-reflecting capacity and the liveliness of their color.

[0038] According to another aspect, the invention relates to a composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K, wherein the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition for use in the treatment of a hair growth disease or hair growth disorder exacerbated by stress.

[0039] Preferably the hair disorder or hair disease is alopecia, more preferably alopecia areata.

[0040] The composition described herein is also suitable for the treatment of forms of alopecia other than alopecia areata, that are exacerbated by stress, for example telogen effluvium, androgenetic alopecia, ringworm and cicatricial alopecia.

[0041] The composition described herein is also suitable for use in other forms of alopecia, temporary or permanent hair loss conditions.

[0042] Specifically, the temporary hair loss conditions comprise the use of the composition in a treatment combination with medicaments or medications that cause stress, such as those due to medication side-effects or hormonal fluctuations.

[0043] The composition finds application also in the treatment of hair growth diseases or hair growth disorders due to stress derived from psychological conditions or acute stress due to a stressful lifestyle or other personal events.

[0044] Advantageously, it was found that the composition described herein, when administered to a subject suffering from thinning or loss of hair, including skin P024192WO-01 Notarbartolo & Gervasi S.p.A. appendages due to stress, causes progressive hair thickening.

[0045] Some aspects of the present invention are illustrated with reference to the accompanying Figures.

[0046] DESCRIPTION OF THE FIGURE

[0047] Figure 1. Expression of the Insuline-Hke growth factor 1 (IGF-1) gene in Human Follicle Dermal Papilla Cells (HFDPC) as determined by RT-PCR. NCTC2544 cells were treated at 37°C for 24h, under 5% CO2, with: basal medium (Positive control); basal medium and Cortisol 50pM (Positive control); Rhodiola rosea extract comprising 1 % of salidroside 0.0033 mg / mL and Cortisol 50pM; Tryptophan 0.0066 mg / mL and Cortisol 50pM; Vitamin K 0.0125 pg / mL and Cortisol 50pM; the combination of Rhodiola rosea extract 0.0033 mg / mL, Tryptophan 0.0066 mg / mL, Vitamin K 0.0125 pg / mL and Cortisol 50pM in complete culture medium. Values are means of 2 experiments in duplicate. *p<0.05; **p<0.01.

[0048] Figure 2. Expression of the tumor necrosis growth factor (TGF-[31 ) gene in Human Follicle Dermal Papilla Cells (HFDPC) as determined by RT-PCR. NCTC2544 cells were treated at 37°C for 24h, under 5% CO2, with: basal medium (Positive control); basal medium and Cortisol 50pM (Positive control); Rhodiola rosea extract comprising 1 % of salidroside 0.0033 mg / mL and Cortisol 50pM; Tryptophan 0.0066 mg / mL and Cortisol 50pM; Vitamin K 0.0125 pg / mL and Cortisol 50pM; the combination of Rhodiola rosea extract 0.0033 mg / mL, Tryptophan 0.0066 mg / mL, Vitamin K 0.0125 pg / mL and Cortisol 50pM in complete culture medium. Values are means of 2 experiments in duplicate. *p<0.05; **p<0.01.

[0049] Figure 3. Expression of the 11 -beta dehydrogenase 1 (Hsd11 b1 ) gene in Human Follicle Dermal Papilla Cells (HFDPC) as determined by RT-PCR. NCTC2544 cells were treated at 37°C for 24h, under 5% CO2, with: basal medium and Cortisol 50pM (Positive control); Rhodiola rosea extract comprising 1 % of salidroside 0.0033 mg / mL and Cortisol 50pM; Tryptophan 0.0066 mg / mL and Cortisol 50pM; Vitamin K 0.0125 pg / mL and Cortisol 50pM; the combination of Rhodiola rosea 0.0033 mg / mL, Tryptophan 0.0066 mg / mL, Vitamin K 0.0125 pg / mL and Cortisol 50pM in complete culture medium. Values are means of 2 experiments in duplicate. *p<0.05; **p<0.01. P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0050] Figure 4. Expression of the Growth Arrest Specific Protein 6 (GAS6) gene in Human Follicle Dermal Papilla Cells (HFDPC) as determined by RT-PCR. NCTC2544 cells were treated at 37°C for 24h, under 5% CO2, with: basal medium and Cortisol 50pM (Positive control); Rhodiola rosea extract comprising 1 % of salidroside 0.0033 mg / mL and Cortisol 50pM; Tryptophan 0.0066 mg / mL and Cortisol 50pM; Vitamin K 0.0125 pg / mL and Cortisol 50pM; the combination of Rhodiola rosea 0.0033 mg / mL, Tryptophan 0.0066 mg / mL, Vitamin K 0.0125 pg / mL and Cortisol 50pM in complete culture medium. Values are means of 2 experiments in duplicate. *p<0.05; **p<0.01.

[0051] DETAILED DESCRIPTION OF THE INVENTION

[0052] The inventors have found that an extract of Rhodiola Rosea, tryptophan and Vitamin K synergistically act by guaranteeing or increasing hair health by preventing or reducing the hair loss, when the extract of Rhodiola rosea and tryptophan are in specific amounts so as to be in a determined ratio.

[0053] Therefore, in a first aspect the invention relates to a composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K, wherein the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition.

[0054] The composition according to the invention, regardless of the cosmetic and / or therapeutic uses, comprises an extract of Rhodiola rosea as biologically active ingredient or component in an amount in the range of 0.0015-1.5 % with respect to the total amount of the composition.

[0055] Rhodiola rosea, often referred to as golden root or Arctic root, is a perennial herb recognized for its adaptogenic properties, which help the body cope with various stressors. This plant has garnered attention for its potential to alleviate stress and enhance cognitive performance, particularly in the context of fatigue and anxiety. The mechanisms through which Rhodiola rosea exerts its effects on stress are diverse, including the modulation of neurotransmitter levels, enhancement of immune function, and reduction of oxidative stress.

[0056] The inventors found out that the extract of Rhodiola rosea acted in a synergistic way with tryptophan and vitamin k on the hair loss when the loss is caused by stress. P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0057] This was surprising since it was not expected that the extract of Rhodiola rosea could act on the hair follicle biology in order to develop effective interventions for stress-related hair loss.

[0058] Preferably the extract of Rhodiola rosea is an extract comprising as key active compound salidroside. The inventors deem that this specific extract was preferred since it could influence neurotransmitter levels such as serotonin and dopamine in an effective way when in composition with vitamin K and tryptophan.

[0059] This neurochemical modulation of the extract makes the composition capable to decrease the perception of stress, making the composition comprising the extract of Rhodiola rosea capable to effectively act on the hair loss of individuals facing high levels of stress.

[0060] Additionally, the composition comprising the extract of Rhodiola rosea had immunomodulatory effects increasing the activity of T-lymphocytes and promoting the secretion of antibodies, which are vital for the immune response. Therefore, the composition of the invention comprising the extract of Rhodiola rosea strengthened immune responses and helped the body manage stress more effectively.

[0061] The inventors were aware that oxidative stress was another critical factor in the body's response to stress, and found out that the composition comprising the extract of Rhodiola rosea in the claimed amounts possessed significant antioxidant properties.

[0062] In conclusion, the composition comprising the extract of Rhodiola rosea aided the body in managing stress through various mechanisms, including the modulation of neurotransmitter levels, enhancement of immune function, reduction of oxidative stress, improvement of physical performance and in the recovery of hair growth.

[0063] The composition according to the invention comprises hence the extract of Rhodiola rosea in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition, preferably the extract of Rhodiola rosea in an amount in the range of 0.03-1.5%, more preferably in the range of 0.3-1.5% with respect to the total amount of the composition.

[0064] The parts used for extraction are mainly and preferably the roots, which are collected, dried and then extracted with a suitable extraction solvent. The plant extract of Rhodiola rosea can be preferably obtained by extraction from plant roots P024192WO-01 Notarbartolo & Gervasi S.p.A. using a physiologically acceptable solvent as an extraction medium. Preferably the extraction solvent is a hydroalcoholic solvent.

[0065] The term “physiologically acceptable solvent” means a solvent that does not produce significant adverse reactions when introduced into the human body or applied to the human body. A suitable solvent for obtaining the plant extract is a physiologically acceptable liquid, in which the biologically active components of the selected plant are soluble and in which they do not undergo an alteration that deprives them of activity.

[0066] In some embodiments, the physiologically acceptable solvent is a protic polar solvent, preferably selected from the group consisting of water, acetic acid, ethanol, n-butanol ethyl acetate, n-butanol, isopropanol, n-propanol, and mixtures thereof. Preferably the extraction solvent is a polar protic solvent selected from the group consisting of water, ethanol, ethyl acetate and mixtures thereof, more preferably it is a mixture of water and ethyl acetate.

[0067] To obtain the plant extract from roots, well-known extraction techniques such as, for example, solid-liquid extraction, can be used to separate / extract one or more biologically active components from the vegetal tissues of the tree.

[0068] Preferably the extract of Rhodiola rosea in the claimed amounts is a hydroalcoholic extract, more preferably comprising the compound salidroside, preferably in an amount from 0.0010 to 10%, preferably 0.003 to 1 .5 %, still more preferably 1 % with respect to the total amount of the extract.

[0069] The composition according to the invention comprises tryptophan as an essential active component in an amount in the range of 0.0030-3 % with respect to the total amount of the composition.

[0070] Tryptophan, an essential amino acid, plays a significant role in the body's response to stress, primarily through its conversion to serotonin, a neurotransmitter that regulates mood, anxiety, and overall emotional well-being. The relationship between tryptophan and stress is complex, involving metabolic pathways that can influence both psychological and physiological responses.

[0071] One of the key pathways through which tryptophan affects stress is the kynurenine pathway. Under conditions of stress, the metabolism of tryptophan can shift from serotonin production towards the formation of kynurenine and its metabolites, which P024192WO-01 Notarbartolo & Gervasi S.p.A. are associated with neurotoxicity and inflammation.

[0072] Surprisingly the composition of the invention comprising tryptophan in the claimed amounts was capable of guaranteeing and increasing this activity, due to the synergistic action with the extract of Rhodiola rosea and vitamin K.

[0073] Without being bound to any theory the inventors found out the right combination of tryptophan with the extract of Rhodiola rosea and vitamin K, when the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition, was capable to manage the relationship between tryptophan, stress, and hair growth, involving metabolic pathways that can significantly influence both psychological and physiological outcomes.

[0074] It was in fact known that under stress the metabolism of tryptophan can shift from serotonin production to the kynurenine pathway, which leads to the formation of kynurenine and its metabolites. This shift is often mediated by increased activity of the enzyme indoleamine 2,3-dioxygenase (IDO), which is upregulated during stress. Elevated levels of kynurenine can contribute to neuroinflammation and oxidative stress, both of which have been associated with hair loss and impaired hair follicle function.

[0075] Therefore a correct combination in the specific amounts as in the composition of the invention allowed to manage this relationship, avoiding the reduction of serotonin levels, which are crucial for maintaining mood and overall well-being, and this reduction can negatively impact hair growth.

[0076] The composition according to the invention comprises hence tryptophan in an amount in the range of 0.0030-3% with respect to the total amount of the composition.

[0077] Preferably tryptophan is in the form of tryptophan L.

[0078] The composition of the invention comprises also Vitamin K.

[0079] It is known that Vitamin K has been implicated in hair growth through its influence on cellular signaling pathways. Research indicates that vitamin K can enhance the expression of Gas6 in dermal papilla cells, which are essential for hair follicle development and cycling. P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0080] Anyway, corticosterone inhibits GAS6 to govern hair follicle stem-cell quiescence. Gas6 promotes the activation of hair follicle stem cells (HFSCs) and supports their transition from a quiescent state to an active growth phase. In conditions of stress, such as elevated corticosterone levels, the expression of Gas6 can be suppressed, leading to prolonged quiescence of HFSCs and subsequent hair loss. Corticosterone inhibits GAS6 to govern hair follicle stem-cell quiescence. Restoring Gas6 expression has been shown to counteract this inhibition.

[0081] Surprisingly the composition of the invention comprising adequate levels of vitamin K and in view of the presence of the extract of Rhodiola rosea and tryptophan, when the extract of Rhodiola rosea is in an amount in the range of 0.0015-1.5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition, generates a synergistic activity, may help maintaining Gas6 expression and supporting hair growth. Furthermore, in view of the presence of vitamin K, the composition by modulating inflammation contributes to guarantee effects on hair health. The composition, by influencing the inflammatory response, may help mitigate stress- induced hair loss. Gas6 itself has been recognized for its protective effects on hair follicles. It interacts with receptor tyrosine kinases, promoting cell survival and preventing apoptosis in hair follicle cells. This protective mechanism is crucial during stress, as elevated stress hormones can lead to increased apoptosis and hair follicle degeneration. The composition of the invention maintains the right relationship between Gas6 and vitamin K and enhances Gas6 signaling, thereby promoting hair follicle health and reducing the risk of hair loss.

[0082] The composition according to the invention comprises preferably Vitamin K in an amount in the range of 0.00015-1.5 % with respect to the total amount of the composition.

[0083] In a further aspect the invention relates to a non-therapeutic, cosmetic use of the composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K, wherein the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition, for preventing or reducing hair loss of a mammal due to stress. P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0084] In the present invention when the term “hair” is used, it is intended to comprise also skin appendages, i.e. beard, eyelashes, eyebrows and nails, of a human being.

[0085] Advantageously, a specific subject of the invention is hence a non-therapeutic, cosmetic use of the composition of the invention in promoting hair growth in a human being.

[0086] In accordance with certain specific aspects of the invention, the hair loss reduction activity of the composition described herein affects a population of individuals who are affected by a greater than physiological hair loss and which can cause thinning on the scalp due to stress.

[0087] The composition of the invention that finds application in the cosmetic field by preventing or reducing the hair loss due to stress allows the achievement of a desired external aspect of hair and skin appendages, such as for example their volume, their shine and / or their compactness, their light-reflecting capacity and the liveliness of their color.

[0088] According to another aspect, the invention relates to a composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K for use in the treatment of a hair growth disease or hair growth disorder exacerbated by stress, wherein the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition.

[0089] Preferably the hair disorder or hair disease is alopecia, more preferably alopecia areata.

[0090] The composition described herein is also suitable for the treatment of forms of alopecia other than alopecia areata, that are exacerbated by stress, for example telogen effluvium, androgenetic alopecia, ringworm and cicatricial alopecia.

[0091] The composition described herein is also suitable for use in other forms of alopecia, temporary or permanent hair loss conditions.

[0092] Specifically, the temporary hair loss conditions comprise the use of the composition in a treatment combination with medicaments or medications that cause stress, such as those due to medication side-effects or hormonal fluctuations.

[0093] The composition finds application also in the treatment of hair growth diseases or hair growth disorders due to stress derived from psychological conditions or acute P024192WO-01 Notarbartolo & Gervasi S.p.A. stress due to a stressful lifestyle or other personal events.

[0094] Advantageously, it was found that the composition described herein, when administered to a subject suffering from thinning or loss of hair, including skin appendages due to stress, causes progressive hair thickening

[0095] In a preferred embodiment for cosmetic and / or therapeutic uses the composition of the invention is formulated with at least one physiologically and / or pharmaceutically acceptable carrier, diluent or excipient.

[0096] The physiologically acceptable carrier of the composition of the invention is an excipient, carrier or diluent suitable for oral application and / or topical administration. The physiologically and pharmacologically acceptable carriers can be the same carriers.

[0097] In the context of the present document, the term “carrier” refers to an excipient, carrier, diluent, or adjuvant that may be present in the composition of the invention. Any carrier and / or excipient suitable for the desired preparation form for administration is contemplated herein.

[0098] Accordingly, the physiologically and / or pharmaceutically acceptable carrier, diluent or excipient may be selected based on the route of administration for which the composition is intended.

[0099] Preferably the composition is formulated for oral administration.

[0100] The compositions for oral administration can be in solid or liquid form.

[0101] Typical compositions in solid form comprise tablets, capsules, powders, granules, pills and gummies. Preferably the composition is formulated as a gummy.

[0102] If desired, the tablets may be coated with a suitable polymeric agent or an agent to achieve sustained / extended release by conventional techniques.

[0103] Exemplary excipients used in solid forms include cellulose derivatives such as hydroxymethyl cellulose, hydroxypropyl methylcellulose, methylcellulose, hydroxypropyl cellulose, hydroxyethyl cellulose, carboxyethyl cellulose, ethylhydroxyethyl cellulose, cellulose acetate butyrate, cellulose acetate phthalate, starch or its derivates and mixtures thereof.

[0104] Other excipients can be sugar alcohols or polyols.

[0105] Further examples of suitable excipients include polymers of the lactam class, typically pyrrolidone and derivatives thereof, such as polyvinylpyrrolidone, P024192WO-01 Notarbartolo & Gervasi S.p.A. polyvinylpolypyrrolidone and mixtures thereof, inorganic salts such as calcium or dicalcium phosphate, lubricants such as magnesium stearate, triacylglycerols and mixtures thereof.

[0106] Tablets, pills, capsules, and the like may also contain a binder such as gum tragacanth, acacia, com starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as com starch, potato starch, alginic acid; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, lactose, saccharin, or other sweeteners. When a unit dosage form is a capsule, in addition to materials of the type indicated above, it may contain a liquid carrier such as a fatty oil.

[0107] In certain embodiments, the composition of the invention further comprises one or more additional components, such as additives, fillers, stabilizers, emulsifiers, structurants, film-forming agents, aromas, plasticizers, wetting agents, and thickeners.

[0108] Various other materials may be present as coatings or to modify the physical shape of the dosage unit. For example, tablets may be coated with shellac, sugar, or both. In addition to the active ingredient, a syrup or elixir may contain sucrose as a sweetening agent, methyl and propylparaben as preservatives, a coloring agent and a flavoring agent such as cherry or orange flavor. To prevent rupture during transit through the upper portion of the gastrointestinal tract, the composition may be an enteric-coated formulation.

[0109] Typical compositions in liquid form for oral administration comprise solutions, suspensions, syrups, emulsions, gels.

[0110] In these forms of administration, suitable carriers include water, glycols, or oil-in- water or water-in-oil emulsions containing the biologically active components dispersed therein.

[0111] In some embodiments, the route of administration of the composition of the invention is topical. The compositions for topical administration can be in solid, semi-liquid or liquid form. Typically, the topical composition is applied to the skin, and especially to the scalp in an effective amount.

[0112] The composition according to the present invention can comprise also one or more excipients suitable for the wished administration formulation. P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0113] The composition of the invention can also comprise other active ingredients / principles.

[0114] The composition of the invention may hence comprise one or more additional ingredients having trichological activity.

[0115] In some embodiments, compositions further comprise additional ingredients such as vitamins, for example vitamin A, vitamin C, additional B vitamins, piroxidine hydrochloridrate (vitamin B6), niacin, and mixtures thereof.

[0116] Preferably the additional ingredients can be shikimic acid, also in form of extract, maslinic acid also in the form of a basil extract.

[0117] Further ingredients that can be present in the composition of the invention are dietary supplements, preferably a dietary supplement of zinc, more preferably zinc gluconate.

[0118] According to a preferred embodiment the composition is administered as a solid oral dosage form, preferably a gummy.

[0119] Suitable excipients that can be cited for preparing a gummy are gelatin, pectin, agar- agar, carrageenan, gelling gum; sweeteners such as maltitol, sorbitol, sucrose, glucose syrup, fructose syrup, xylitol, erythritol; edible colorants; natural and artificial flavors; modified starches; maltodextrin; gums; solvents such as glycerin, propylene glycol and other known excipients.

[0120] Typically, the active ingredients of the composition are formulated in dosage units. The dosage unit may contain from 0.1 to 1000 mg of active ingredient per dosage unit of the finished product for daily administration.

[0121] In accordance with some embodiments, a dosage form comprises the extract of Rhodiola rosea in an amount in the range from 0.005 mg to 500 mg, preferably from 0.5 to 50 mg per dose, more preferably 10 mg.

[0122] In accordance with some embodiments, a dosage form comprises tryptophan in an amount in the range from 0.005 mg to 500 mg, preferably from 0.5 to 60 mg per dose, more preferably 20 mg.

[0123] In accordance with some embodiments, a dosage form comprises Vitamin K in an amount in the range from 0.005 mg to 500 mg, preferably from 0.05 to 60 mg per dose, more preferably 75pg. P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0124] For oral uses, a cosmetically or pharmaceutically active amount of the composition of the invention is administered preferably twice a day.

[0125] The amount administered and the frequency of administration of the composition for use in medical field will depend on the type and severity of the tricholog ical condition to be treated.

[0126] The invention will be detailed in the following examples to better illustrate the advantages of the composition of the invention.

[0127] EXPERIMENTAL PART

[0128] EXAMPLE 1 The ingredients in the amount indicated in Table 1 were used for preparing gummies with further excipients needed to prepare a gummy to have a gummy of about 3000 mg.

[0129] Table 1 EXAMPLE 2

[0130] The ingredients in the amount indicated in Table 2 were used for preparing a topical liquid composition to be contained in a vial.

[0131] Table 2 P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0132] EXAMPLE 3

[0133] Assessment of the synergic activity of Rhodiola rosea extract comprising 1% salidroside, Tryptophan and Vitamin K (composition of the invention)

[0134] PromoCell® Human Follicle Dermal Papilla Cells (HFDPC) were used in order to test the hair growth promoting activity of Rhodiola Rosea extract, tryptophan and Vitamin K.

[0135] Human follicle dermal papilla cells are found in the dermal papillae at the base of hair follicles. These cells are essential for induction and maintenance of hair growth. Since these cells express androgen receptors, they represent a well-suited model system for androgen-related research.

[0136] PromoCell® Human Follicle Dermal Papilla Cells (HFDPC) were purchased by PromoCell. The cell line was cultured in a specific medium (follicle dermal papilla cell growth medium-ready to use with its supplement mix, PromoCell) in conditions of complete sterility and maintained at 37°C with 5% carbon dioxide (CO2) atmosphere.

[0137] HFDPC are isolated principally from normal human dermis of the lateral scalp. Shortly after isolation, all Human Follicle Dermal Papilla Cells are cryopreserved at passage 2 (P2) using proprietary serum-free freezing medium, Cryo-SFM.

[0138] Every two days confluent cultures were splitted 1 :3-1 :6, after washing with phosphate buffered saline (PBS) (without Ca2+and Mg2+), using trypsin / EDTA, and seeded at a density of 1 x106cells per well into 12-well plates for qRT-PCR.

[0139] All the compounds were tested in single and in combination.

[0140] Following concentrations were used:

[0141] 1 . Negative control: HFDPC in complete culture medium

[0142] 2. Rhodiola rosea extract 0.0033 mg / mL in complete culture medium P024192WO-01 Notarbartolo & Gervasi S.p.A.

[0143] 3. Tryptophan 0.0066 mg / mL in complete culture medium

[0144] 4. Vitamin K 0.0125 pg / mL

[0145] 5. Rhodiola rosea estract 0.0033 mg / mL + Tryptophan 0.0066 mg / mL + Vitamin K 0.0125 pg / mL in complete culture medium

[0146] Twenty-four hours after seeding on 12-well plates, HFDPC 80% confluent cells were exposed to compounds taking into account the ratio at which the single compounds in the botanical extracts, respectively.

[0147] RNA for qRT-PCR analysis was extracted 24h after the treatment. Tri Reagent (Sigma Aldrich) methods were used and the cDNA was then synthesized from 2pg RNA template in a 20pl reaction volume, using the PrimeScript RT-PCR Kit (Takara, Japan). The cDNA was amplified and detected by the Stratagene Mx3000P Real- Time PCR System (Agilent Technologies Italia S.p.A., Milan, Italy).

[0148] The amplification of cDNA from NCTC cells was conducted using the following Taqman gene expression assays: Hs01547870_m1 (Hsd11 b1 ), Hs01547656_m1 (IGF-1 ), Hs00155410_m1 (TGF- [31 ), and Hs999999 ml (human glyceraldehyde-3- phosphate dehydrogenase, GAPDH). GAPDH was used as housekeeping gene.

[0149] PCR amplifications were carried out in a 20pl of total volume. The mixture of reaction contained 10pl of 2X Premix Ex Taq (Takara, Japan), 1 pl of 20 TaqMan gene expression assay, 0.4 pl of RoX Reference Dye II (Takara, Japan), 4.6 pl of water and 4pl of DNA. PCR conditions were the following: 95°C for 30 sec followed by 40 cycles of 95°C for 5 sec, 60°C for 20 sec. PCR reactions were performed in duplicate using an MX3000p PCR machine (Stratagene, La Jolla, CA). A cycle threshold. (34] Surprisingly, as can be seen in the results shown in Figure 1 , the combination of Rhodiola rosea extract 0.0033 mg / mL, Tryptophan 0.0066 mg / mL and Vitamin K 0.0125 pg / mL showed a higher significant stimulatory effect on IGF-1 than the compounds tested alone (same concentration).

[0150] Contextually, the combination of Rhodiola rosea 0.0033 mg / mL, Tryptophan 0.0066 mg / mL and Vitamin K 0.0125 pg / mL showed a higher inhibitory effect on TGF-[31 gene than the compound tested alone (same concentration) (Figure 2).

[0151] Surprisingly, the combination of Rhodiola rosea 0.0033 mg / mL, Tryptophan 0.0066 mg / mL and Vitamin K 0.0125 pg / mL showed a significant (p<0.05) inhibitory effect on the hydroxysteroid 11 -beta dehydrogenase 1 (Hsd11 b1 ) gene and the effect was P024192WO-01 Notarbartolo & Gervasi S.p.A. higher than the compound tested alone (same concentration) (Figure 3). When active, the enzyme can regulate the negative effects of glucocorticoids on DPCs.

[0152] High levels of corticosterone / stress suppress Gas6 and prevent the transition from late telogen to anagen prolonging late telogen. Most interesting, the combination of Rhodiola rosea 0.0033 mg / mL, Tryptophan 0.0066 mg / mL and Vitamin K 0.0125 pg / mL showed a significant (p<0.05) positive effect on the expression of Growth

[0153] Arrest Specific Protein 6 (GAS6) gene and the effect was higher than the compound tested alone (same concentration) (Figure 4).

[0154] Taking together, the data on the expression of IGF-1 , TGF-[31 , Hsd11 b1 and GAS6 genes suggested a strong hair growth promoting effect of the combination of

[0155] Rhodiola rosea extract 0.0033 mg / mL, Tryptophan 0.0066 mg / mL and Vitamin K 0.0125 pg / mL and this effect is synergic.

Claims

P024192WO-01 Notarbartolo & Gervasi S.p.A.CLAIMS1. A composition comprising an extract of Rhodiola Rosea, tryptophan and Vitamin K, wherein the extract of Rhodiola rosea is in an amount in the range of 0.0015-1 .5 % with respect to the total amount of the composition and tryptophan is in an amount in the range of 0.0030-3 % with respect to the total amount of the composition.

2. The composition according to claim 1 , wherein the extract of Rhodiola rosea comprises the compound salidroside in an amount from 0.0015 to 10 %, preferably about from 0.003 to 1 .5%, more preferably about 1 % with respect to the total amount of the extract.

3. The composition according to claim 1 or claim 2, wherein tryptophan is in the form of tryptophan L.

4. The composition according to anyone of claims 1 -3, wherein Vitamin K is in an amount in the range of 0.00015-1.5 % with respect to the total amount of the composition.

5. A non-therapeutic, cosmetic use of the composition according to anyone of claims 1 -4 for preventing or reducing hair loss of a mammal due to stress.

6. The non-therapeutic cosmetic use of claim 5, wherein the mammal is a human being.

7. A composition according to anyone of claims 1 -4 for use in the treatment of a hair growth disease or hair growth disorder exacerbated by stress.

8. The composition for use according to claim 7, wherein the hair growth disorder or hair growth disease is alopecia, preferably selected from the group consisting of alopecia areata, telogen effluvium, androgenetic alopecia, ringworm and cicatricial alopecia.

9. The non-therapeutic cosmetic use of anyone of claims 5-6 and the composition for use of anyone of claims 7-8, wherein the composition is formulated for the oral administration or the topical administration.

10. The non-therapeutic cosmetic use of claim 9, wherein the composition is orally administered in the form of gummy.

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