Long-term treatment of alopecia

Abstract

The present invention relates to a topical composition for use in the treatment and / or prevention of alopecia in a subject, wherein the therapeutic effect lasts for at least 24 weeks or more after the last administration of the composition. Kits and methods for treating and / or preventing alopecia using the compositions are also disclosed.

Description

TECHNICAL FIELD

[0001] The present invention relates to compositions, preferably topical compositions, for use in the treatment and / or prevention of hair loss in a subject, wherein the therapeutic effect lasts for at least 24 weeks or more after the last administration of the composition. Kits and methods of using the compositions for the treatment and / or prevention of hair loss are also disclosed. BACKGROUND

[0002] Hair follicles (HFs) are unique micro-organs that undergo a continuous, life-long regeneration cycle. The lower part of a healthy anagen HF (bulge and hair bulb) enjoys relative immune privilege (IP), which protects the follicle from inflammatory processes and promotes immune tolerance. These unique HF compartments are characterized by factors that act as IP sentinels to maintain HF IP (Lintzeri, 2022).

[0003] As the hair follicle develops, blood vessels originating from the deep dermal vascular plexus surround the follicle. These blood vessels nourish the follicle and support nutrient transport, waste removal, and growth (Murphey, 2022). The vasculature controls the guidance of mobile immune cells, thereby supporting the protective immune function that protects our body from pathogens and the like. In case of inflammation or immune surveillance, cells arranged at the luminal side of the blood vessels, called endothelial cells (ECs), attract and guide mobile immune cells to the appropriate exit sites in the vasculature, allowing the cells to enter the underlying tissue. Thus, “immune-regulatory ECs” (IMECs) (Amersfoort, 2022) fulfill an important supportive role in the guidance and directional migration of immune cells in transport. During inflammation, ECs expose a variety of adhesion molecules on their surface, which slow down and arrest the movement of immune cells in the blood circulation. These adhesion molecules are thought to provide the immune cells with a guidance cue to cross the blood vessel wall through a multi-step process called transendothelial migration (TEM) or diapedesis (Schimmel, 2017).

[0004] In a normal hair growth cycle, only sporadic immune cells can be found around the hair bulb of anagen HFs, and very occasionally within the hair bulb of anagen HFs (Lintzeri, 2022). But in Alopecia Areata (AA), genetic or exogenous factors cause significant unwanted CD8+ T cell transendothelial migration (TEM), especially from the vasculature to the hair follicle. 8+ T cell transendothelial migration (TEM).

[0005] Histologically, AA lesions exhibit a unique, dense perifollicular and intrafollicular inflammatory cell infiltrate around the hair bulb region, resembling a swarm of bees, forcing HFs prematurely into catagen, dystrophy, and ultimately apoptosis. CD8+ T cells are often the first cells to penetrate into the intrafollicular location, severely disrupting HF integrity (Lintzeri, 2022).

[0006] During AA flares, the anagen phase of the hair growth cycle is significantly shortened, leading to an acute onset of scarring alopecia, ranging from small, well-demarcated patchy areas on the scalp to complete loss of scalp and body hair. While the exact etiology of AA is not fully elucidated, it is recognized that HF hair bulb IP collapse plays a key role in the pathophysiology of the disease. Exactly what causes this IP collapse is not fully understood.

[0007] Local inflammation in AA is primarily mediated by the JAK-STAT pathway. In AA, there is overexpression of proinflammatory cytokines that signal through their receptors via the JAK / STAT pathway. This leads to JAK-mediated production of IFN-g and IL-15, which in turn promotes an inflammatory feedback loop that further promotes local inflammation. Given the key role of the JAK-STAT pathway in mediating CD8+ NKG2D+ T cell responses, a major component of AA pathogenesis, it is not surprising that JAK inhibitor drugs, which inhibit the JAK enzymes, interfere with the JAK-STAT signaling pathway, thereby blocking downstream signaling of different cytokines, thereby protecting the hair follicle from damage and the start of a new anagen phase and hair regrowth (Dillon, 2021).

[0008] The first JAK inhibitor was approved for the treatment of AA in 2022, representing the first approved treatment for this debilitating chronic autoimmune disease, and more are in development (Dillon, 2021). Since the first JAK inhibitor was approved for the treatment of rheumatoid arthritis in 2011 (Shawky, 2022), real-world evidence of serious health-related risks has been gathered (Hoisnard, 2022), leading to a black box warning from the US Food and Drug Administration and the European Medicines Agency for the entire class of JAK inhibitors (Kragstrup, 2022).

[0009] Due to the serious risks associated with JAK inhibitor immunosuppression, a timely drug holiday is necessary.

[0010] However, since JAK inhibitors "only" block the JAK-STAT signaling pathway, treatment cessation systemically leads to rapid relapse of the disease within 3 months and only a full relapse (Askin, 2021). Tissue-resident memory T cells (TRM) are long-lived lymphocytes that reside in tissues and develop after the initiation of a T cell-mediated immune response. Indeed, endothelial cells are obligate interaction partners for T cell trafficking into inflamed tissues and thus can drive the development of TRM. Wienke et al. have shown that in a co-culture system of human cytokine-activated ECs and FACS-sorted T cells, the interaction of T cells with activated ECs primes for tissue residency (Wienke, 2022). The relapse of AA, the demonstrated upregulation of TRM cells, and the reported clinical benefit of JAK inhibitors during administration all support a key role of TRM cells in the pathogenesis of the disease (Ryan, 2021).

[0011] Prior to the first drug approval for AA, the standard of care consisted mainly of off-label use of corticosteroids in addition to the previous immunosuppressants (Meah, 2020). Unfortunately, corticosteroids, especially topical corticosteroids (TCS), are documented to cause potential local side effects, including impairment of epidermal barrier function and skin atrophy (Wollenberg, 2018). Moreover, transdermal absorption of TCS through a broken skin can lead to systemic exposure and subsequent growth impairment (Coureau, 2008). Due to actual and perceived side effects and potential long-term toxicity, people often have poor compliance with TCS due to "corticophobia" (Stalder, 2017).

[0012] Likewise, due to the severe risks associated with corticosteroids, a timely drug holiday is necessary. However, similar to JAK inhibitors, disease relapse after treatment cessation is the norm (Lintzeri, 2021).

[0013] Systemic disease relapse after treatment cessation puts the life of AA patients into a demoralizing "emotional rollercoaster" (McGettigan, 2013), the rhythm of which is that hair can regrow for a while after treatment and then fall out for a while after treatment cessation.

[0014] There is a high unmet medical need for a treatment that is both safe enough and does not require a drug holiday and that is effective after treatment interruption. SUMMARY

[0015] This object is achieved by providing a composition for use in the treatment and / or prevention of alopecia in a subject, said composition comprising as active ingredient an effective amount of an Allium species (Allium species ) extract, Citrus spp. Citrus species ) extract, Acanthopanax spp. Paullinia species ) aqueous extract and Theobroma spp. Theobroma species ) extract, wherein the composition is applied topically for a period of time necessary to detect one or more therapeutic effects consisting of slowing hair loss, stimulating hair growth and / or increasing hair density, and wherein the one or more therapeutic effects last for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks or more after the last application of the composition, the therapeutic effects being evidenced by measuring the Severity of Alopecia Tool (SALT) score.

[0016] It is a further object of the present application to provide a method for treating and / or preventing hair loss in a subject, the method comprising administering a composition comprising, as active ingredients, an effective amount of extract of Allium spp., extract of Citrus spp., extract of Acanthopanax spp. and extract of Theobroma spp., wherein the composition is applied topically for a period of time necessary to detect one or more therapeutic effects consisting of slowing hair loss, stimulating hair growth and / or increasing hair density, and wherein the one or more therapeutic effects last for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks or more after the last application of the composition, the therapeutic effects being evidenced by measuring the Severity of Alopecia Tool (SALT) score.

[0017] It is a further object of the present application to provide the use of the composition of the present application for the manufacture of a medicament for treating and / or preventing hair loss in a subject.

[0018] It is a further object of the present application to provide a kit for treating and / or preventing hair loss, the kit comprising the composition of the present application or the composition for use of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 Example of SALT score (adapted from Olsen, 2004).

[0020] Figure 2 The SALT score of the patients treated with the composition continued to improve and throughout the follow-up the change in SALT score was continuous and statistically significantly better than placebo. DETAILED DESCRIPTION

[0021] Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The publications and applications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present application is not entitled to antedate such publication. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0022] In the event of conflict, the present specification, including definitions, controls. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this subject matter belongs. As used herein, the following definitions are provided for ease of reference.

[0023] The terms "comprise" (or comprise), "comprising," (or comprising) and "comprises" (or comprises) are generally used in the sense of "including" (or including), i.e., permitting the presence of one or more features or components. The term "comprise" (or comprise), "comprising," (or comprising) and "comprises" (or comprises) also includes the more restrictive terms "consist of" and "consisting essentially of," respectively.

[0024] As used in the specification and claims, the singular form "a," "an" and "the" include plural references unless the context clearly dictates otherwise.

[0025] As used herein, the terms "subject" / "subject in need thereof or "patient" / "patient in need thereof are art-recognized and used interchangeably herein to refer to a mammal, including dogs, cats, rats, mice, monkeys, cows, horses, goats, sheep, pigs, camels, and most preferably, humans. In some instances, the subject is a subject in need of treatment or a subject suffering from a disease or disorder. In other aspects, however, the subject can be a normal subject. The term does not denote a particular age or sex. Adult and newborn subjects, whether male or female, are therefore intended to be encompassed by this term. Preferably, the subject is a human, most preferably, a human suffering from hair loss or who can be at risk of suffering from hair loss.

[0026] The term "about," particularly with reference to a given quantity, number, or percentage, means a deviation of plus or minus ten percent (+10%). For example, about 5% includes any value between 4.5% and 5.5%, such as 4.5, 4.6, 4.7, 4.8, 4.9, 5, 4.1, 5.2, 5.3, 5.4, or 5.5.

[0027] As used herein, "at least one" means "one or more", "two or more", "three or more", and the like. For example, at least 8 weeks means 8 weeks or more, i.e. 9 weeks, 10 weeks, 11 weeks, and the like.

[0028] In the context of the treatment and prevention of alopecia, such as alopecia areata (AA), the inventors surprisingly found that the therapeutic effect of the composition of the present invention persists for several weeks or even months after the last administration.

[0029] It is known that AA recurs after existing treatments have stopped (Lintzeri-2022).

[0030] The inventors found that the use of the composition of the present invention comprising aqueous extract of Paullinia cupana produces a more potent anti-inflammatory effect, which is an unexpected synergistic effect produced together with the other components in the composition. Without being bound by theory, the use of aqueous extract of Paullinia cupana allows to obtain more caffeine than with aqueous-alcoholic extraction. The introduction of more caffeine into the composition by aqueous extraction, in contrast to aqueous-alcoholic extraction, unexpectedly produces this potent anti-inflammatory effect together with the other components in the composition, achieving an unexpected synergistic effect.

[0031] The inventors believe that the administration of the composition of the present invention not only inhibits the activation of ECs, improves endothelial permeability, but also unexpectedly leads to the persistence of the therapeutic effect after treatment interruption (i.e. long-term effect after treatment stop).

[0032] In one aspect, the present invention provides a composition comprising, as active ingredients, an effective amount of extract of Allium species, extract of Citrus species, aqueous extract of Paullinia cupana and extract of Theobroma species, wherein the composition is topically administered for a period of time necessary to detect one or more therapeutic effects consisting of slowing down hair loss, stimulating hair growth and / or increasing hair density, and wherein the one or more therapeutic effects persist after the last administration of the composition, which is evidenced by measuring the Severity of Alopecia Tool (SALT) score.

[0033] In one aspect, the one or more therapeutic effects persist for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks or more.

[0034] The composition according to the present invention can be prepared according to any conventional method known in the art.

[0035] The term “treatment” refers to any administration of the compositions, pharmaceutical compositions, therapeutic agents, active ingredients, compounds, etc. of the present disclosure to a subject for the following purposes: (i) inhibiting the disease, i.e., arresting the development of clinical symptoms; (ii) reversing the disease, and / or (iii) relieving the disease, i.e., causing the regression of clinical symptoms.

[0036] The term “prevention” as used herein refers to any administration of the compositions, pharmaceutical compositions, therapeutic agents, active ingredients, compounds, etc. of the present disclosure to a subject for the purpose of preventing the disease, i.e., for the purpose of not developing clinical symptoms and signs of the disease.

[0037] In the context of the present invention, the disease is alopecia. In one aspect, the alopecia is an immune-mediated disease, preferably an autoimmune disease. More preferably, the autoimmune disease is alopecia areata.

[0038] Alopecia areata (AA) is an immune-mediated autoimmune inflammatory hair disease affecting pediatric and adult patients. The immune inflammatory attack of the scalp hair follicle aborts the normal hair cycle, leading to its premature entry into telogen phase, and to hair loss. The disease starts with the appearance of perifollicular inflammation and immune inflammatory cell invasion around the growing hair hair bulb, leading to breakdown of HF immune privilege, tissue dystrophy, HF cell death, and hair shaft shedding (Lintzeri-2022). NKG2D + T cells, natural killer (NK) cells, and autoreactive CD8+ T lymphocytes play a key role, which recognize self-antigens upon MHCI, MHC II expression exposure on follicular cells. T cell activation is accompanied by an increase in IFN-g secretion, recruiting or activating more inflammatory cells (including macrophages, mast cells, and dendritic cells), leading to accelerated cell death and apoptosis of hair follicle cells (Bertolini, M., 2020).

[0039] The early clinical presentation of AA is manifested as random patchy hair loss, which can sometimes lead to total scalp hair loss (alopecia totalis), or even to spread throughout all body sites (alopecia universalis). Patients affected by AA come from all races, but are more common in females than in males, both with similar clinical presentation. Most importantly, when AA starts early in infancy, it is usually more severe in prognosis and associated with higher frequency of unpredictable relapses in adulthood (Villasante Fricke, A.C., 2015).

[0040] As used herein, the term "effective amount" means, within the bounds of reasonable medical judgment, a therapeutically effective amount of the compositions, pharmaceutical compositions, formulations, therapeutic agents, active ingredients, compounds, etc. of this disclosure, which is high enough to significantly and positively alter the symptoms and / or condition to be treated, but low enough to avoid serious side effects (at a reasonable risk / benefit ratio).

[0041] In the context of this invention, the compositions of this invention comprise effective amounts of Allium species extract, Citrus species extract, Aquatic extract of Cercis chinensis species, and Cocoa species extract as active ingredients.

[0042] The term Allium genus extract or water-alcohol extract specifically refers to extracts from all species of the Allium genus (Liliaceae family), particularly onion (Onion). Allium cepa The water-alcohol extract and natural extract obtained from )

[0043] The term "citrus species extract" or "water-alcohol extract" specifically refers to extracts from all species of the genus Citrus (Rutaceae), particularly lemon (Citrus spp.). Citrus limon The water-alcohol extract and natural extract obtained from )

[0044] The term "extract from the genus *Awakening Vine*" (nebulized or non-nebulized) or its aqueous extracts specifically refers to extracts from the genus *Awakening Vine* (family Sapindaceae). Sapindaceae All species, especially guarana ( Paullinia cupana Water extracts and natural extracts obtained from ).

[0045] The term "cocoa extract" (atomized or non-atomized) or "water-alcohol extract" specifically refers to extracts from the genus *Coccus* (family Malvaceae). Malvaceae All species, especially cocoa trees ( Theobroma cacao The water-alcohol extract and natural extract obtained from )

[0046] In one aspect of the invention, a period of time is required for the composition of the invention or the composition used in the invention to be topically applied (typically to the external skin surface of the skull) to detect one or more therapeutic effects, said therapeutic effects including slowing hair loss, stimulating hair growth and / or increasing hair density, or consisting of slowing hair loss, stimulating hair growth and / or increasing hair density.

[0047] The time required to detect one or more therapeutic effects typically includes about 16 weeks to about 48 weeks, preferably about 20 weeks to about 40 weeks, more preferably about 20 weeks to about 32 weeks, and even more preferably about 24 weeks.

[0048] Any method known in the art can be used to detect, monitor, and / or demonstrate the effects of one or more treatments, which consist of slowing hair loss, stimulating hair growth, and / or increasing hair density.

[0049] In one aspect, the one or more therapeutic effects are detected, monitored and / or evidenced by measuring a Severity of Alopecia Tool (SALT) score.

[0050] The SALT score is a calculation based on a scoring system. The scalp is divided into four regions: left scalp, which accounts for 18% of the scalp surface area; right scalp, which accounts for 18% of the scalp surface area; top, which accounts for 40% of the scalp surface area; and back, which accounts for 24% of the scalp surface area. The percentage of hair loss in each of the four scalp regions is determined independently, each multiplied by its coefficient (left and right: 0.18 each; top: 0.40; back: 0.24). The coefficient for each region varies depending on the location. The resulting percentages of hair loss for all four regions are then added together as the final total percentage of hair loss, designated as the SALT score.

[0051] Olsen et al. also describe a SALT score assessment that is more suitable for clinical trials. According to the scalp region, each region is further subdivided into four quadrants (5%+4%+4%+5%; 10%+10%+10%+10% or 6%+6%+6%+6%), see Figure 1 .

[0052] The percentage of hair loss in each quadrant is determined independently, each multiplied by its coefficient (0.04, 0.05, 0.06 or 0.1). The coefficient for each quadrant varies depending on the location (see Figure 1 ). The resulting percentages of hair loss for all four regions (16 quadrants) are then added together as the final total percentage of hair loss, designated as the SALT score.

[0053] Since obtaining the final SALT score requires a significant amount of calculation, and since the variation of the coefficient depends on the region, there is a risk of error with a manual SALT score. Therefore, in one aspect, the SALT score is computer-implemented. In a preferred aspect, the SALT score assessment described by Olsen is used to detect, monitor and / or evidence the one or more therapeutic effects.

[0054] Typically, the subject’s baseline SALT score is determined prior to initiating administration of the composition of the application.

[0055] In one aspect, a decrease in the detected SALT score of at least about 2% or more, at least about 5% or more, at least about 10% or more, at least about 15% or more, at least about 20% or more, at least about 30% or more, at least about 40% or more, or at least about 50% or more, when compared to the subject’s baseline SALT score, indicates that one or more therapeutic effects are detected and that administration of the composition of the application is effective.

[0056] A decrease in SALT score of at least about 2% or more, at least about 5% or more, at least about 10% or more, at least about 15% or more, at least about 20% or more, at least about 30% or more, at least about 40% or more, or at least about 50% or more detected when compared to the subject's SALT score determined after the last administration of the composition indicates that one or more therapeutic effects are detected and persist after the topical administration is discontinued. In some aspects, the one or more therapeutic effects persist, and even improve.

[0057] Generally, the compositions of the present application are topically administered at least once a day, at least twice a day, or more, preferably topically administered to the external skin surface of the skull.

[0058] In related aspects, the compositions of the present application are formulated for topical administration. Preferably, the composition is mixed with a diluent and / or excipient to formulate for topical administration containing about 20% or more by weight of the composition based on the total weight of the composition.

[0059] Generally, a volume of about 0.5 ml to 2.5 ml of the compositions of the present application formulated for topical administration is administered at least once a day, at least twice a day, or more, to cover the entire scalp of the subject.

[0060] In one aspect, the compositions of the present application or the compositions for use according to the present application comprise: about 65% to about 93% by weight of water-alcohol extract of Allium species, about 5% to about 33% by weight of water-alcohol extract of Citrus species, about 0.25% to about 2.5% by weight of water extract of Polygala species, and about 0.25% to about 2.5% by weight of water-alcohol extract of Theobroma species.

[0061] In one preferred aspect, the compositions of the present application or the compositions for use according to the present application comprise: about 65% to about 93% by weight of water-alcohol extract of Allium cepa, about 5% to about 33% by weight of water-alcohol extract of Citrus limon, about 0.25% to about 2.5% by weight of water extract of Paullinia cupana, and about 0.25% to about 2.5% by weight of water-alcohol extract of Theobroma cacao.

[0062] In one even more preferred aspect, the compositions of the present application or the compositions for use according to the present application comprise: about 87% by weight of water-alcohol extract of Allium cepa, about 12% by weight of water-alcohol extract of Citrus limon, about 0.67% by weight of water extract of Paullinia cupana, and about 0.67% by weight of water-alcohol extract of Theobroma cacao.

[0063] In one aspect, the composition of the present application or the composition for use according to the present application further comprises an excipient. Examples of excipients include about 0.05 wt% to about 8.0 wt% sodium chloride and about 1 wt% to about 40 wt% glycerin, based on the total weight of the composition.

[0064] In one aspect, the composition of the present application or the composition for use according to the present application comprises about 0.05 wt% to about 8.0 wt%, preferably about 0.1 wt% to about 7.0 wt%, more preferably about 0.4 wt% to about 6.0 wt%, even more preferably about 0.9 wt% to about 3 wt% sodium chloride, based on the total weight of the composition.

[0065] In one aspect, the composition of the present application or the composition for use according to the present application comprises about 1 wt% to about 20 wt%, preferably about 1.2 wt% to about 15 wt%, more preferably about 1.8 wt% to about 10 wt%, even more preferably about 2 wt% to about 5 wt% glycerin, based on the total weight of the composition.

[0066] Compositions of the present application suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin, such as solutions, lotions, shakes, creams, ointments, gels, foams, transdermal patches, powders, solids, sponges, strips, vapors, pastes, tinctures, microparticles, microcapsules, nanoparticles, liposomes, or emulsions. Preferably, the composition of the present application suitable for topical administration is in the form of a solution or a lotion.

[0067] The present application also contemplates a method of treating and / or preventing hair loss in a subject.

[0068] In one aspect, the method of treating and / or preventing hair loss in a subject comprises: administering a composition comprising, as active ingredients, an effective amount of an extract of Allium species, an extract of Citrus species, an extract of Polygala species, and an extract of Theobroma species, wherein the composition is administered topically for a period of time necessary to detect one or more therapeutic effects consisting of slowing hair loss, stimulating hair growth, and / or increasing hair density, and wherein the one or more therapeutic effects last for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks, or more after the last administration of the composition, the therapeutic effects being evidenced by measuring the Severity of Hair Loss Tool (SALT) score.

[0069] The present application also contemplates the use of the composition of the present application in the manufacture of a medicament for treating and / or preventing hair loss in a subject, comprising administering the medicament comprising, as active ingredients, an effective amount of an extract of Allium species, an extract of Citrus species, an extract of Polygala species, and an extract of Theobroma species, wherein the medicament is applied topically for a period of time required to detect one or more therapeutic effects consisting of slowing hair loss, stimulating hair growth, and / or increasing hair density, and wherein the one or more therapeutic effects persist for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks, or more after the last application of the composition, the therapeutic effects evidenced by measuring a Severity of Alopecia Tool (SALT) score.

[0070] The present application also contemplates kits for use in the treatment and / or prevention of alopecia as described herein. In one aspect of the application, the kit comprises a composition of the present application or a composition for use as described in the present application.

[0071] The kits of the present application can further include a container and a label or package insert on or associated with the container. Suitable containers include, for example, bottles, vials, syringes, dispensers, spray devices, and the like. The containers can be formed from a variety of materials such as glass or plastic.

[0072] The label or package insert can include instructions for use. The instructions included can be affixed to the container, or can be included as a package insert. While the instructions are typically written materials or printed materials, they are not limited thereto. Any medium capable of storing such instructions and communicating them to the end user is contemplated by the present disclosure.

[0073] The present disclosure should be considered in all respects as illustrative and not restrictive, the scope of the application being indicated by the appended claims, and all changes which come within the meaning and range of equivalency are intended to be embraced therein.

[0074] Examples Materials and Methods Composition The composition comprises about 87% by weight of water-alcohol extract of Allium cepa, about 12% by weight of water-alcohol extract of Citrus limon, about 0.67% by weight of water extract of Paullinia cupana, and about 0.67% by weight of water-alcohol extract of Theobroma cacao.

[0075] Topical formulation The final topical solution product comprises at least 20% of the above composition. The remainder of the finished product consists of water and other conventional excipients in skin solutions such as betaine, glycerol, sodium chloride, and the like.

[0076] A final skin solution containing 22.25% of the composition (LH-8) has been evaluated in a clinical trial assessing the safety and efficacy of the composition in subjects with AA (RAAINBOW study, ClinicalTrials.gov Identifier NCT03240627). About 1 mL of the composition was used on the entire scalp, twice a day, with an interval of about 12 hours (e.g., in the morning and in the evening).

[0077] The RAAINBOW study was a double-blind, randomized, multicenter study (versus placebo). 107 subjects (male and female) participated and were analyzed for safety. Of these, 62 subjects, who met the inclusion criteria (i.e., moderate to severe AA), were analyzed for efficacy, representing the Full Analysis Set (FAS) population.

[0078] Results Below is a summary of the safety analysis.

[0079]

[0080] No subject in the composition group experienced a serious adverse event (AE). Only 5.6% of the subjects treated with the composition experienced an AE that was possibly related to the drug (11.1% for the placebo group). One AE was severe, consisting of severe scalp and facial eczema. However, eczema is the most common comorbidity of AA, and patients with AA are more likely to have atopic dermatitis, eczema (17.4% vs. 2.2% control) (Conic, 2020). All other AEs were mild, moderate, local, and transient.

[0081] No side effects observed with JAK inhibitors and corticosteroids were reported in this study.

[0082] In addition, the RAAINBOW study was specifically designed to assess the efficacy of the composition on AA after 6 months of treatment, and also to assess the potential disease relapse after treatment discontinuation (measured 6 months after discontinuation).

[0083] AA was assessed by the scalp alopecia severity score (which is called SALT (Scalp Alopecia Severity Tool) score) based on global standardized scalp photographs. The SALT score was developed by Olsen in 2004 (Olsen, 2004) “to help facilitate well-controlled clinical trials for alopecia.” It consists of a standardized method to assess the degree of hair loss, with a 100% SALT score being completely bald. It is based on a 4-picture-based scoring system, where each side of the head is divided into 4 quadrants and scored accordingly by the investigator (see Figure 1). The SALT score is a standard measurement endpoint used in all trials evaluating the efficacy of new treatments for AA. An example is presented below.

[0084] In the RA INBREED study, the SALT score was measured at baseline (V1), after 3 months of treatment (V2) and after 6 months of treatment (V3) Figure 2 ). Treatment was then discontinued and patients were re-evaluated after 3 months of treatment-free follow-up (V4) and after 6 months of treatment-free follow-up (V5).

[0085] After checking the assumption of normality of the data (using Q-Q plot and Shapiro-Wilk test), the generalized linear model (GLIMMIX procedure in SAS) was preferred to the MMRM. The GLIMMIX model had as dependent variable the rank of the relative change in SALT score from baseline, as fixed effects the treatment, visit, treatment-visit interaction and as covariate the baseline severity based on SALT score.

[0086] There was a statistically significant difference between the LH-8 group and the placebo group in terms of both the relative change and the absolute change in SALT score from baseline to week 24, with the LH-8 group showing greater change. Thus, the planned primary endpoint was met (p-value: <0.0001).

[0087] Among the subjects treated with the composition who improved during the 6-month treatment period (between V1 and V3), only 4% of the subjects relapsed after treatment discontinuation, as measured after the 6-month treatment-free period (V5). 96% of the subjects who responded to treatment did not relapse after treatment discontinuation. Furthermore, as shown in Figure 2 , the SALT score of the patients treated with the composition continued to improve and the change in SALT score was continuous and statistically significantly better than placebo throughout the follow-up period.

[0088] In chronic conditions, a period of 6 months is considered to represent the evolution of the disease (Ostbye, 2005). Thus, a 6-month treatment-free follow-up period can be considered to represent the long-term evolution of the disease Figure 2 ).

[0089] In the present case, we can conclude that, except for 4% of the cases, discontinuing treatment with the composition does not lead to a relapse of the disease. Thus, unlike JAK inhibitors, corticosteroids and despite the safety that makes discontinuation unnecessary, treatment with the composition can be discontinued without the risk of a relapse of the disease. While AA is a chronic condition that requires long-term treatment, the composition allows treating AA not in a long-term manner, but only until the disease is controlled, after which treatment can be safely interrupted.

[0090] References

[0091]

Claims

1. A composition for use in treating and / or preventing hair loss in a subject, comprising effective amounts of Allium species extract, Citrus species extract, Aquatic extract of Cercis chinensis species and Cocoa species extract as active ingredients; in, The composition is applied topically for a period of time during which one or more therapeutic effects are detected, the therapeutic effects consisting of slowing hair loss, stimulating hair growth, and / or increasing hair density; Furthermore, the therapeutic effect of one or more of the said treatments lasts for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks, or longer after the last application of the composition, and the therapeutic effect is demonstrated by measuring the Severity of Hair Loss Tool (SALT) score.

2. The composition for use according to claim 1, wherein the hair loss is an immune-mediated disease.

3. The composition for use according to claim 2, wherein the immune-mediated disease is an autoimmune disease.

4. The composition for use according to claim 3, wherein the autoimmune disease is alopecia areata.

5. The composition for use according to any one of the preceding claims, wherein the time required to detect one or more therapeutic effects includes about 16 weeks to about 48 weeks, preferably about 20 weeks to about 40 weeks, more preferably about 20 weeks to about 32 weeks, and even more preferably about 24 weeks.

6. The composition for use according to any one of the preceding claims, wherein the one or more therapeutic effects are detected or demonstrated by measuring the Severity of Hair Loss Tool (SALT) score.

7. The composition for use according to any one of the preceding claims, wherein when compared with the baseline SALT score of the subject determined before the administration of the composition, a decrease in SALT score of at least about 2% or more, at least about 5% or more, at least about 10% or more, at least about 15% or more, at least about 20% or more, at least about 30% or more, at least about 40% or more, or at least about 50% or more is detected.

8. The composition for use according to any one of the preceding claims, wherein, When compared with the subject's SALT score determined after the last administration of the composition, a decrease in SALT score of at least about 2% or more, at least about 5% or more, at least about 10% or more, at least about 15% or more, at least about 20% or more, at least about 30% or more, at least about 40% or more, or at least about 50% or more was detected.

9. A composition for use according to any one of the preceding claims, wherein the composition comprises: about 65% to about 93% by weight of an aqueous-alcoholic extract of an Allium species, about 5% to about 33% by weight of an aqueous-alcoholic extract of a Citrus species, about 0.25% to about 2.5% by weight of an aqueous extract of a Caulis Aristolochia, and about 0.25% to about 2.5% by weight of an aqueous-alcoholic extract of a Cocoa species.

10. A composition for use according to any one of the preceding claims, wherein the composition comprises: about 65% to about 93% by weight of an aqueous-alcoholic extract of onion, about 5% to about 33% by weight of an aqueous-alcoholic extract of lemon, about 0.25% to about 2.5% by weight of an aqueous extract of guarana, and about 0.25% to about 2.5% by weight of an aqueous-alcoholic extract of cocoa tree.

11. The composition for use according to claim 10, wherein the composition comprises: about 87% by weight of an aqueous-alcoholic extract of onion, about 12% by weight of an aqueous-alcoholic extract of lemon, about 0.67% by weight of an aqueous extract of guarana, and about 0.67% by weight of an aqueous-alcoholic extract of cocoa.

12. A composition for use according to any one of the preceding claims, wherein the composition is mixed with a diluent and / or excipient to formulate a composition suitable for topical application containing about 20% by weight or more based on the total weight of the composition.

13. A composition for use according to any one of the preceding claims, wherein, It also contains, as an excipient, about 0.05% to about 8.0% by weight of sodium chloride and about 1% to about 40% by weight of glycerol, based on the total weight of the composition.

14. The composition for the use according to claim 13, wherein, Based on the total weight of the composition, the composition contains about 0.05% to about 8.0% by weight, preferably about 0.1% to about 7.0% by weight, more preferably about 0.4% to about 6.0% by weight, and even more preferably about 0.9% to about 3% by weight of sodium chloride.

15. The composition for the use according to claim 13 or 14, wherein, Based on the total weight of the composition, the composition contains about 1% to about 20% by weight, preferably about 1.2% to about 15% by weight, more preferably about 1.8% to about 10% by weight, and even more preferably about 2% to about 5% by weight of glycerol.

16. A composition comprising, as an active ingredient, an effective amount of an extract of an Allium species, an extract of a Citrus species, an aqueous extract of a Caulis Aristolochia species, and an extract of a Caulis Cocoa species, and at least one excipient.

17. The composition of claim 16, wherein the composition is mixed with a diluent and / or excipient to formulate a composition suitable for topical application containing about 20% by weight or more based on the total weight of the composition.

18. A method of treating and / or preventing hair loss in a subject, comprising administering the composition of any one of claims 1 to 17, The time required for the composition to be detected after topical application, wherein the therapeutic effect comprises slowing hair loss, stimulating hair growth, and / or increasing hair density. And among them, The one or more therapeutic effects last for at least 8 weeks, at least 12 weeks, at least 16 weeks, at least 20 weeks, at least 24 weeks or longer after the last application of the composition, and the therapeutic effects are demonstrated by measuring the Severity of Hair Loss Tool (SALT) score.

19. Use of the composition of any one of claims 1 to 17 in the preparation of a medicament for treating and / or preventing hair loss in a subject.

20. A kit for treating and / or preventing hair loss, comprising the composition of claim 16 or 17, or the composition for use as described in any one of claims 1 to 15.

21. The kit of claim 20, further comprising one or more containers, and a label or packaging insert on or associated with said one or more containers.

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