Garcinia mangostana extract for promoting hair growth

Abstract

To provide a composition for promoting hair growth, applicable in the fields of cosmetics and dermatology.SOLUTION: Provided is a dermatological composition for use in the prevention or treatment of alopecia by promoting hair growth, the composition comprising at least one hydroalcoholic extract of Garcinia mangostana and at least one dermatologically acceptable excipient.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a mangosteen (Garcinia mangostana L.) extract and a composition comprising said extract for promoting hair growth, in particular for stimulating hair regrowth, with applications in the fields of cosmetics and dermatology. [Background technology]

[0002] Hair care, not only for cosmetic purposes but also to prevent hair loss and to regenerate hair, is a constant subject of research. In an attempt to determine the etiology of hair loss in cases of baldness, alopecia, etc., many theories attribute it to seborrhea, increased tension of the tissue on the cranial sphere, reduced blood supply, or some kind of endocrine or nervous disorder.

[0003] Hair follicles are small organs anchored in the subcutaneous tissue of the skin and their main function is the generation of hair shafts. Their distribution is established during fetal development and their number is genetically determined. Hair follicles are dynamic structures that generate hair in a cycle of tissue growth and remodeling. This cycle can be divided into three stages: · Growth phase (anagen): Cells in the dermal papilla (fibroblasts) send signals to stem cells in the hair bulb, which causes them to multiply. The cells transform to envelop the dermal papilla and form the hair's sulfur matrix. They divide and differentiate into follicular keratinocytes, the cells that give the hair its structure. To provide the hair's structure, these keratinocytes require sulfur proteins, vitamin B6, and various minerals (e.g. zinc and magnesium). The duration of this phase determines the length of the hair and depends on the proliferation and differentiation of matrix cells at the base of the hair follicle. Phase of regression (catagen): The matrix dies, so that the hair papilla is no longer in contact with this matrix. There is no more exchange between cells. The hair follicle and the hair papilla return to the epidermis. · Resting phase (telogen): The cells of the hair papilla and hair bulb are intact and inactive. The hair falls out. The cycle must be restarted in order for a new hair to grow.

[0004] Therefore, hair is constantly being renewed, and most of the 100,000 to 150,000 hairs that make up the scalp are in the growth phase. In a healthy scalp, normal and physiological hair loss occurs at about 60 to 100 hairs per day. Any further hair loss, whether occasional or constant, is said to be a disease.

[0005] The term alopecia refers to partial or total hair loss. Alopecia can be related to many factors, such as genetic factors, age, sex, illness, stress, hormonal disorders, side effects of medications, scars. Several forms of alopecia can be distinguished: Hereditary androgenetic alopecia is the most common and causes premature hair loss in genetically predisposed individuals, particularly men. It manifests as reduced hair volume and even baldness, affecting 50% of men over the age of 50. Postmenopausal alopecia is the most common cause of baldness in women. Hair loss is more diffuse and widespread in women than in men. Diffuse alopecia in women is a disorder that often develops after menopause and affects approximately 40% of women over the age of 70. The term diffuse refers to the fact that, unlike in men, hair loss occurs evenly across the scalp. Acute or reactive alopecia may be associated with medications, stress, childbirth, severe nutritional deficiencies, iron deficiency, and hormonal disorders and is the simultaneous, diffuse loss of large amounts of hair. Cicatricial alopecia can be caused by skin diseases (tumours, burns, abrasions), acute irradiation, lupus erythematosus or parasites (tinea, lichen). Alopecia areata is thought to be autoimmune in origin and is characterized by a more or less large patchy attack in one or more areas. Congenital alopecia is rare and corresponds to the absence of a hair follicle or an abnormality (mutation) of the hair.

[0006] Alopecia is essentially associated with a disturbance in hair renewal, resulting in an acceleration of cycle frequency, at the expense of first the quality of hair, then its quantity. The most frequent phenomenon is a shortening of the duration of the growth (anagen) phase, accompanied by the cessation of cell proliferation. The result is an early induction of catagen and an increase in the number of hair follicles in the telogen phase, and thus an increase in hair loss. Therefore, to combat alopecia, it is necessary to restart the hair cycle (e.g. by activating the anagen phase).

[0007] To date, various products have been proposed for the treatment of alopecia, in particular for inducing or promoting hair growth. Many of them combine several active ingredients that may have a beneficial effect on the biological parameters involved in hair loss. Among the most commonly used active ingredients, we can mention by way of example vitamins, such as vitamins A, E, B5, B6, C, H and PP; trace elements, such as zinc, copper, magnesium, silicon; protein derivatives, such as peptides, sulfur-containing amino acids (e.g. methionine, cystine, cysteine ​​or derivatives); lipophilic or hydrophilic plant essential oils or extracts, but these are not limited to them; antifungal agents, such as piroctone olamine, undecycline derivatives, ciclopirox olamine; chemically synthesized molecules known to act specifically on the androgen receptor level or on the activity of 5α-reductase. Minoxidil (i.e. 2,4-diamino-6-piperidinopyrimidine-3-oxide) is today applied in the treatment of androgenetic alopecia. Its mechanism of action has not been clearly elucidated, despite many theories.Moreover, in many clinical cases, even if stabilization of hair loss is observed, its effectiveness is limited, because as soon as treatment is stopped, resumption of alopecia process is observed.Its restriction of daily use is considered to be the cause of undesirable side effects seen in patients who use it for a long time, such as local skin reaction or systemic effects.

[0008] Furthermore, for hair restoration, compositions containing a wide range of active ingredients have been proposed, these active ingredients being, for example, 2,4-diaminopyrimidine-3-oxide derivatives as described in patent application EP0522964. Clinical studies have shown that PGF2α analogues have hair and eyelash growth inducing properties in humans and animals (Johnstone, Am J Opht, 124(4), 544-547, 1997). In humans, tests carried out on the scalp have shown that the prostaglandin E2 analogue Viprostol has hair density increasing properties. Patent WO98 / 33497 describes pharmaceutical compositions containing prostaglandins or prostaglandin derivatives for promoting hair growth.

[0009] Thus, despite the many options currently available, consumers still need new products to promote hair regrowth that are as effective as chemical active ingredients, yet are natural and environmentally friendly.

[0010] Recently, researchers (Harel et al., Sci. Adv. 2015; e1500973) found that applying drugs to the skin that inhibit enzymes of the Janus kinase (JAK) family promotes rapid and dense hair growth in experiments carried out on mouse and human hair follicles. Two JAK inhibitors have already been approved by the US Food and Drug Administration (FDA), one called rusolitinib for the treatment of blood disorders and the other tofacitinib for the treatment of rheumatoid arthritis. Both of them are the subject of clinical trials for alopecia areata. These studies confirmed that topical application of JAK inhibitors to mouse skin resulted in hair regrowth, showing that they not only thwart autoimmune attacks but also act on hair follicles to bring them out of the telogen phase. Mice treated with either of the two JAK inhibitors for five days showed new hair regrowth within 10 days, indicating a rapid acceleration of the growth process, whereas control mice showed no hair regrowth during the same period.

[0011] Furthermore, mangosteen is a tropical tree native to Southeast Asia that is now cultivated in many tropical countries for its edible fruit, mangosteen, also known as the "fruit of the gods" or "queen of fruits."

[0012] Mangosteen is a dioecious tree that can reach a height of 20 m in the wild. The leaves are smooth, glossy, leathery, elliptic to elliptic-oblong, 14-25 cm by 5-10 cm. They are cuneate to subcircular at the base with a short, pointed apex. Male flowers are rare and occur in groups of 2-9 at the ends of twigs; female flowers are slightly larger than the male flowers and occur singly or in pairs. The fruit consists of a thick pericarp that is pink to very dark, tinged with purple at maturity, contains edible white pulp and encases 4-5 seeds.

[0013] Mangosteen has been known for centuries in Asian traditional medicine for its antioxidant, anti-inflammatory and antibacterial properties. The peel, in particular, is used for skin infections, diarrhea, abdominal pain, urinary tract infections or bruises. It is also used for fevers (Ovalle-Magallanes et al., Food Chem Toxicol 109:102-122, 2017). There has also been a huge craze for mangosteen in the West over the last decade. There are many juices based on the pulp and peel, dietary supplements and topical preparations containing mangosteen on the market, and recent clinical studies have shown benefits in various indications, such as weight loss (Udani et al., Nutr J,8:48, 2009) and periodontitis (Mahendra et al., J Investig Clin Dent,8(4),2017). Mangosteen extracts are also found as active ingredients in some cosmetic products for anti-aging or slimming effects.

[0014] The mangosteen peel is an important source of xanthones, the main ones being α-mangosteen and γ-mangosteen (Ovalle-Magallanes et al., 2017). In addition, mangosteen peel contains tannins, anthocyanins and sugars. Fruit extracts, especially those of mangosteen peel more or less enriched in xanthones, have been the subject of many pharmacological studies, both in vitro and in vivo. The main actions demonstrated are antitumor (especially in the prostate, lung, breast and colon), anti-inflammatory, antioxidant, antidiabetic, antihyperlipidemic and antibacterial.

[0015] Mangosteen extracts combined with other extracts to form specific compositions for topical use have been the subject of several patent applications in the field of hair science. Patent application JP2016069334 relates to mangosteen extracts combined with hydrolyzed silk in liquid form, positioning it as a hair protectant against UV rays. Patent application CN103735453 relates to a composition for dyeing hair. Patent application US2006210515 relates to a topical composition containing partially hydrolyzed fucoidan, which may be useful for hair regrowth. Natural ingredients, such as mangosteen, can be added to this composition, but there is no activity related to this in the field of hair science. A Thai study aimed to screen plant extracts that have 5α-reductase inhibitory activity and are therefore useful in the treatment of benign prostatic hyperplasia and / or hereditary alopecia. An ethanol extract of mangosteen peel was screened, but no relationship between the content of total phenolic compounds and 5α-reductase inhibitory activity was established. The 5α-reductase inhibitory activity of mangosteen extracts is also the subject of two patent applications. Application JP2000229857 attributes the 5α-reductase inhibitory activity to xanthones, including α-mangosteen. Xanthones can be extracted and isolated from plants of the families Hypericaceae (Guttiferae), Gentianaceae, Mulberry, Polygonaceae, Liliaceae, and Plumageae. The preparation of a benzene extract of mangosteen is described. Application JP5017365 relates to several plant extracts, including aqueous extracts of mangosteen or extracts obtained with hydrophilic organic solvents (e.g. methanol extracts), which can be formulated for topical application.

[0016] Thus, to date, there is no bibliographical data stating that mangosteen extracts may have activity in hair regrowth. Summary of the Invention

[0017] Surprisingly, the inventors have found that mangosteen (Garcinia mangostana L.) extract has the desired pharmacological activity for promoting hair growth, in particular for promoting hair regrowth, by acting on JAK targets. These activities are detailed in Examples 11-13.

[0018] The present invention therefore relates to the treatment of hair loss insofar as mangosteen (Garcinia mangostana L.) extract acts on the functional or biological mechanisms at the origin of hair growth.

[0019] More particularly, the present invention relates to a hydroalcoholic extract of mangosteen (Garcinia mangostana) for use in preventing or treating alopecia by promoting hair growth, and to a dermatological composition comprising said extract and at least one dermatologically acceptable excipient.

[0020] The present invention also relates to the non-therapeutic use of a hydroalcoholic extract of mangosteen (Garcinia mangostana) for promoting hair regrowth.The present invention also relates to a cosmetic composition for promoting hair regrowth, in particular a cosmetic composition for promoting hair regrowth comprising at least one hydroalcoholic extract of mangosteen (Garcinia mangostana) and at least one dermatologically acceptable excipient. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] definition In the present invention, the plant Garcinia mangostana L. may be abbreviated by the term Garcinia mangostana.

[0022] "Mangosteen (Garcinia mangostana) extract" refers to an extraction product of all or part of the mangosteen plant.

[0023] "Extract" means the product obtained after extraction of a part of a plant with a solvent called the extractant, i.e. the product present in the extractant (which may be in concentrated or dried form after evaporation of part or all of the extractant). The extract may be a dry extract.

[0024] In the present invention, "dry extract" refers to an extract that does not contain an extracting solvent or carrier, or that contains only an insignificant trace amount of an extracting solvent or carrier. Thus, such a dry extract contains only substances derived from mangosteen. Also, the dry extract may contain an insignificant trace amount of an extracting solvent.

[0025] In the present invention, "about" means that the value in question may be 10%, particularly 5%, and especially 1% lower or higher than the stated value. This definition applies in particular when defining the weight content of α-mangostin based on the weight of the dry extract.

[0026] "Hydrophilic solvent" means in particular a solvent selected from the group consisting of water, subcritical water, water-miscible alcohols such as ethanol, C3-C5 glycols, glycerol, acetone, and mixtures thereof.

[0027] In the present invention, "non-polar solvent" means a solvent selected from, for example, heptane, hexane, limonene, halogenated hydrocarbons (chloroform, dichloromethane), supercritical CO2, a mixture of supercritical CO2 and ethanol.

[0028] In the present invention, "moderately polar solvents" refers in particular to solvents selected from the group consisting of C1-C5 alcohols, glycols (such as propylene glycol, butylene glycol, butanediol or pentylene glycol), glycerol, acetone, alkyl esters (such as ethyl acetate, isopropyl acetate, etc.), water-miscible solvents (such as hydroalcohol mixtures or mixtures of acetone and water). This group also includes alternative solvents of hydrotropic type (water-soluble amphiphilic molecules which, from a sufficient concentration, can extract moderately polar compounds as described in the characterization of the extract).

[0029] "Head and body hair" means head hair, body hair, eyebrows, eyelashes and / or fur, preferably hair.

[0030] "Skin appendage" means head hair, body hair, eyebrows, eyelashes and / or nails, preferably hair.

[0031] "Alopecia" refers to the complete or partial loss of scalp and / or body 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 alopecia, scarring alopecia, alopecia areata and congenital alopecia. The result of alopecia is the temporary or permanent, partial or complete absence of scalp and / or body hair.

[0032] "Treating" alopecia means stopping, reducing and / or alleviating alopecia. Thus, "treating" alopecia includes limiting scalp and / or body hair loss and / or promoting scalp and / or body hair growth and / or increasing hair follicle density and / or regulating the phase of the hair follicle cycle.

[0033] "Preventing" alopecia means reducing the risk of the occurrence of, or slowing the progression of, alopecia in a mammal, preferably a human, susceptible to developing alopecia.

[0034] "Limit" means to slow, reduce, decrease, and / or stop.

[0035] "Promote" means to increase, intensify, advantage, amplify, and / or accelerate.

[0036] In the present invention, "cosmetically or dermatologically acceptable" means something that is useful in the preparation of a cosmetic or dermatological composition and is generally safe, non-toxic, free from undesirable toxic properties, biologically or otherwise, and acceptable for cosmetic or dermatological use, in particular for topical application to the hair and / or scalp.

[0037] "Topical application" means application to the skin, in particular the scalp, mucous membranes and / or skin appendages, in particular the hair and scalp.

[0038] Detailed Description of the Invention According to a first aspect, the present invention relates to a mangosteen extract for use to promote hair growth, in particular to promote hair regrowth.

[0039] In the present invention, the mangosteen extract is obtained from one or more parts of the mangosteen plant selected from the above-ground parts (eg, fruit, fruit skin, fruit pulp, seeds, leaves, stems and / or bark, etc.).

[0040] In a particular embodiment of the invention, the mangosteen extract is obtained from the mangosteen fruit and / or from the pericarp of the mangosteen fruit. Advantageously, the mangosteen extract is obtained from the pericarp of the mangosteen fruit.

[0041] In a particular embodiment of the present invention, the mangosteen extract is obtained from a culture of mangosteen cells.

[0042] The mangosteen extract is a hydroalcoholic extract, in particular a hydroethanolic extract.

[0043] According to a preferred embodiment, the extract according to the invention can be obtained by the method described below.

[0044] The mangosteen plant or plant parts may be fresh, dried, whole, cut or crushed and then subjected to the extraction process.

[0045] The method for preparing the extract according to the present invention comprises the steps of extracting all or part of the mangosteen plant with a mixture of alcohol and water.

[0046] In one embodiment of the present invention, the extraction solvent is a mixture of alcohol and water, and the alcohol may be a C3-C5 glycol or a C1-C5 alcohol.

[0047] Advantageously, the mixture of alcohol and water is characterized in that the alcohol / water ratio is between 9:1 and 7:3 (v / v), and even more advantageously, the mixture of alcohol and water is characterized in that the alcohol / water ratio is 9:1 (v / v).

[0048] Advantageously, the mixture of alcohol and water is a mixture of ethanol and water.

[0049] More advantageously, the mixture of ethanol and water is characterized in that the ethanol / water ratio is between 9:1 and 7:3 (v / v).

[0050] More advantageously, the mixture of ethanol and water is characterized in that the ethanol / water ratio is 9:1 (v / v).

[0051] According to another particular embodiment of the invention, the extraction is carried out under stirring or statically, at reflux, at room temperature or at a temperature between room temperature and reflux. The extraction can be assisted by ultrasound, microwave, flash expansion or extrusion, for 1 min to 48 h, with a ratio of plant weight to volume of solvent that can vary from 1:3 to 1:30. The extraction can be repeated 2 to 3 times.

[0052] According to another particular embodiment of the invention, the pomace is then separated from the extract by centrifugation or filtration in order to recover a clear, particle-free liquid phase. The liquid phase contained in the extract can be concentrated to a greater or lesser extent until a dry extract is obtained.

[0053] In another embodiment of the invention, a carrier may be added during the concentration step to obtain an extract containing 1-75% dry extract. The carrier may be maltodextrin, lactose, silica, glycerin, glycol, vegetable oil, hydrotrope, mixture of water and solubilizer, mixture of water and surfactant or any other cosmetically acceptable carrier that solubilizes the extract, preferably of biological origin, such as biobased glycols (1,2-pentanediol; 1,3-butanediol; 1,3-propanediol, etc.), esterified fatty acids and hydrotropes such as alkylglycosides (Sepiclear, Apyclean, APXC4, etc.).

[0054] According to a particular embodiment of the invention, the mangosteen extract may be decolorized, for example with activated carbon. Advantageously, the mangosteen extract is not decolorized.

[0055] The hydroalcoholic extract of mangosteen according to the present invention induces inhibition of the JAK-STAT signaling pathway.Furthermore, the hydroalcoholic extract of mangosteen according to the present invention inhibits melanin synthesis.

[0056] According to a second aspect, the present invention relates to a dermatological or cosmetic composition for use to promote hair growth, in particular to promote hair regrowth, comprising at least one mangosteen extract and at least one dermatologically or cosmetically acceptable excipient.

[0057] Advantageously, the extracts comprised in the dermatological or cosmetic composition are as described above.

[0058] According to a particular embodiment of the present invention, the cosmetic or dermatological composition comprising at least one mangosteen extract as described above and at least one cosmetically or dermatologically acceptable excipient comprises 0.001 to 5 wt. %, more preferably 0.002 to 2 wt. %, more preferably 0.005 to 1 wt. %, and even more preferably 0.01 to 0.5 wt. % of mangosteen extract by weight of dry extract, based on the total weight of the composition.

[0059] Preferably, the present invention relates to a cosmetic or dermatological composition according to the invention, which is in a form suitable for topical application, in particular for topical application to the scalp and / or hair.

[0060] The cosmetic or dermatological compositions according to the invention may thus be in the forms usually known for topical administration, i.e. in particular lotions, shampoos, balms, foams, gels, dispersions, emulsions, sprays, serums, masks or creams, and may in particular use excipients allowing penetration, in order to improve the properties and the accessibility of the active ingredients.

[0061] Advantageously, the compositions according to the invention may be in the forms normally known for topical administration to the hair and scalp, i.e. in particular as a shampoo, a conditioner, a hair cream, a hair lotion, a pack or a leave-in spray.

[0062] A distinction is made between formulated products that can be rinsed and formulated products that do not need to be rinsed.

[0063] Preferably, the composition according to the invention has a light texture that allows optimal penetration without making the hair and / or body hair, nor the scalp, oily.

[0064] In a particular embodiment of the invention, the composition according to the invention is characterized in that it is in a form suitable for oral administration.

[0065] According to another embodiment of the present invention, the composition according to the present invention may be in a form generally known for oral administration, namely, in particular, tablets, capsules, powders, granules, and oral solutions or suspensions.When preparing a solid composition in tablet form, the main active ingredient is mixed with a pharmaceutical carrier such as gelatin, starch, lactose, magnesium stearate, talc, gum arabic, silica, etc.The tablet may be coated with sucrose or other suitable materials, and may be processed in such a way that the activity is extended or delayed, and a predetermined amount of active ingredient is continuously released.

[0066] Capsules can be obtained by mixing the active ingredient with a diluent and pouring the resulting mixture into soft or hard capsules.

[0067] The compositions according to the invention, for example those administered topically or orally, generally contain, in addition to the extracts described above, a physiologically acceptable medium, generally a water-based or solvent-based medium, such as an alcohol, ether or glycol, and may also contain surfactants, complexing agents, preservatives, stabilizers, emulsifiers, thickeners, gelling agents, humectants, emollients, trace elements, essential oils, fragrances, dyes, moisturizers, geothermal water, etc.

[0068] From the very first step of administering the composition according to the invention, for example topically or orally, the hair regains strength and vitality.

[0069] Thus, according to the first or second aspect of the invention, the extract as defined above or the composition as defined above can be used to promote hair growth, in particular to promote hair regrowth.

[0070] In certain embodiments, the mangosteen extract described above is the sole active ingredient for promoting hair growth, and in particular for promoting hair regrowth.

[0071] According to a particular embodiment of the present invention, the above-mentioned mangosteen extract is characterized by an α-mangosteen content of 0.5 to 80%, in particular 5 to 80%, more in particular 10 to 60%, and even more in particular about 15%, based on the weight of the dry extract.

[0072] According to one embodiment of the present invention, the above-mentioned mangosteen extract or the above-mentioned cosmetic or dermatological composition comprising at least one mangosteen extract and at least one cosmetically or dermatologically acceptable excipient is intended for topical and / or oral application, preferably for topical application.

[0073] The cosmetic or dermatological extracts or compositions described above can be used by individuals who have received hair micrografts.

[0074] According to a particular embodiment of the present invention, the mangosteen extract as described above or the composition as described above prevents or treats alopecia, which may be selected from the group consisting of male pattern baldness, reactive alopecia, postmenopausal alopecia and alopecia areata.

[0075] The subject of the present invention is a mangosteen extract as described above or a dermatological composition as described above comprising at least one mangosteen extract and at least one dermatologically acceptable excipient, for use in preventing and / or treating alopecia, which can be selected from the group consisting of reactive alopecia and alopecia areata.

[0076] Another object of the present invention is the use of a mangosteen extract as defined above, or of a dermatological composition as defined above comprising at least one mangosteen extract and at least one dermatologically acceptable excipient, for the manufacture of a pharmaceutical or dermatological composition for promoting hair growth, in particular for the prevention and / or treatment of alopecia, such as reactive alopecia and alopecia areata.

[0077] The present invention also relates to the use of a mangosteen extract as described above, or of a dermatological composition as described above comprising at least one mangosteen extract and at least one dermatologically acceptable excipient, for promoting hair growth, in particular for promoting hair regrowth, in particular for preventing and / or treating alopecia, such as reactive alopecia and alopecia areata.

[0078] The present invention also relates to a method for promoting hair growth, in particular for preventing and / or treating alopecia, such as reactive alopecia and alopecia areata, which comprises administering to a patient in need thereof an effective amount of the mangosteen extract described above, or of a cosmetic or dermatological composition described above comprising at least one mangosteen extract and at least one cosmetically or dermatologically acceptable excipient.

[0079] A subject of the present invention is also the (non-therapeutic) use of said extract according to the above-mentioned embodiments or of said cosmetic or dermatological composition according to the above-mentioned embodiments as a cosmetic for the care of the hair and / or the scalp and / or for promoting hair growth, in particular for promoting hair regrowth and / or for increasing the density of hair follicles and / or for obtaining more covering hair and / or for promoting hair regrowth and / or for treating alopecia, which can be selected from androgenetic alopecia and postmenopausal alopecia.

[0080] The subject of the present invention is also a non-therapeutic cosmetic method for the care of the hair and / or the scalp and / or for promoting hair growth, in particular for promoting hair regrowth and / or for increasing the density of hair follicles and / or for obtaining a higher hair coverage and / or for promoting hair follicle regrowth and / or for treating alopecia, which can be chosen from male pattern baldness and postmenopausal alopecia.

[0081] Another subject of the present invention is a non-therapeutic cosmetic method for promoting hair growth, in particular for promoting hair regrowth, in particular for preventing and / or treating alopecia, such as androgenetic alopecia and postmenopausal alopecia, which comprises using a mangosteen extract as defined above or a cosmetic or dermatological composition as defined above comprising at least one mangosteen extract and at least one cosmetically or dermatologically acceptable excipient. EXAMPLES

[0082] The following examples illustrate the invention without limiting its scope.

[0083] The first ten examples relate to the preparation of extracts for use in the present invention.

[0084] Example 1: Reflux extraction with 90% ethanol 377 g of crushed dried mangosteen peel was extracted with 3.8 L of ethanol and water mixture with ethanol / water ratio of 90:10 (v / v) in a reactor under reflux and stirring for 1 h. The extract was then filtered through a K900 filter plate and the solvent was evaporated to obtain 100 g of orange powder with a mass yield of 26%. The obtained dry extract contains 15.9% α-mangosteen by weight.

[0085] Example 2: Reflux extraction with 90% ethanol and post-treatment 200 g of crushed dried mangosteen peel was extracted with 2 L of ethanol and water mixture with ethanol / water ratio of 90:10 (v / v) in a reactor under reflux and stirring for 1 hour. The extract was then filtered through K900 filter disc and dried with maltodextrin to obtain 142 g of extract as orange-beige powder. The resulting extract contains 75% maltodextrin and 3.5% α-mangosteen based on the weight of the extract.

[0086] Example 3: Reflux extraction with 90% ethanol and post-treatment 1000 g of crushed dried mangosteen peel was extracted with 10 L of ethanol and water mixture with ethanol / water ratio of 90:10 (v / v) in a reactor under reflux and stirring for 1 hour. The extract was then filtered through K900 filter plate and dried with 1,2-pentanediol to obtain 747 g of extract in the form of a dark viscous liquid. The extract obtained contains 70% 1,2-pentanediol and 4.9% α-mangosteen based on the weight of the extract.

[0087] Example 4: Reflux extraction with hexane 17 g of crushed dried mangosteen peel was extracted with 170 mL of hexane in a reactor under reflux for 1 hour. The extract was then filtered through a K900 filter plate and the solvent was evaporated to obtain 200 mg of orange-yellow paste with a mass yield of 1.2%. The resulting dried extract contained 75.0% by weight of α-mangosteen.

[0088] Example 5: Ultrasonic extraction using 96% ethanol 36 g of crushed dried mangosteen peel was contacted with 350 mL of 96% ethanol and extracted under the action of ultrasound (20 kHz) at 100% amplitude, 3 times per minute. The extract was filtered through a K900 filter plate and the solvent was evaporated to obtain 7.9 g of purple-red powder with a mass yield of 22%. The obtained dry extract contained 16.4% by weight of α-mangosteen.

[0089] Example 6: Reflux extraction with 96% ethanol 20 g of crushed dried mangosteen peel was extracted with 200 mL of 96% ethanol under reflux in a reactor for 1 hour. The extract was then filtered through a K900 filter plate and the solvent was evaporated to obtain 5.3 g of purple-red powder with a mass yield of 26%. The resulting dry extract contained 16.0% α-mangosteen by weight.

[0090] Example 7: Hydrotropic extraction with 1.5M aqueous heptyl glucoside solution 26 g of crushed dried mangosteen peel was extracted with 260 mL of 1.5 M aqueous heptyl glucoside solution at 40° C. for 2 h under stirring. After filtration through a K900 filter plate, the filtrate was acidified to pH=2 and diluted with 11 volumes of acidified water at pH=2. After centrifugation, the pellet was collected and dried to give an orange paste with a mass yield of 7.3%. The obtained dry extract contains 17.0% α-mangostin by weight.

[0091] Example 8: Hydrotropic extraction with 25% butyl xyloside aqueous solution 20 g of crushed dried mangosteen peel was extracted with 200 mL of 25% by weight butyl xyloside aqueous solution at room temperature for 2 hours with stirring. After filtration through a K900 filter plate, the filtrate was diluted with 2 volumes of water. After centrifugation, the pellet was collected. The supernatant was diluted with 2 volumes of water. After centrifugation, the pellet was collected and combined with the previous pellet to obtain a brown powdery mangosteen dry extract with a mass yield of 4.9%. The obtained dry extract contains 53.0% by weight of α-mangostin.

[0092] Example 9: Extrusion-assisted hydrotropic extraction using aqueous butyl xyloside solution (APXC4) 670 g of dried mangosteen peel was introduced into the first barrel of a Clextral BC45 twin-screw extruder at a flow rate of 40 kg / h. Then, a 26% by weight aqueous solution of butyl xyloside was introduced at a flow rate of 120 kg / h. The temperature applied to the different barrels was 60° C. After 1 minute, the mangosteen peel extract was collected at the outlet of the extruder through a filtering barrel that allows solid-liquid separation. After clarification, the solution was diluted with 2 volumes of water. After centrifugation, a pellet was collected. The supernatant was diluted with 2 volumes of water. After centrifugation, the pellet was collected and combined with the previous pellet to obtain an orange-coloured, pasty mangosteen dry extract with a mass yield of 0.4%. The obtained dry extract contains 50.0% by weight of α-mangosteen.

[0093] Example 10: Supercritical CO 2 Extraction by 470 g of crushed dried mangosteen peel was extracted with supercritical CO2 at a flow rate of 10 kg / h at 50°C and 50 bar for 2 hours. The extract was then solubilized in ethanol, filtered through a K900 filter plate, and the solvent was evaporated to obtain 3.4 g of orange-yellow paste with a mass yield of 0.7%. The resulting dry extract contained 9.45% α-mangosteen by weight.

[0094] Then, 466 g of the pomace was extracted with supercritical CO2 and co-solvent ethanol (flow rates of 10 kg / h and 1 kg / h, respectively) at 50°C and 50 atm for 1 h. The extract was filtered through a K900 filter disc and the solvent was evaporated to give 2.3 g of orange-yellow paste with a mass yield of 0.5%. The resulting dry extract contained 22.3% α-mangostin by weight.

[0095] Example 11: Evaluation of the inhibitory effect of mangosteen extract on human JAK1, JAK2 and JAK3 JAK-STAT is a signaling pathway that controls growth, survival, differentiation and pathogen resistance. This pathway mediates the effects of cytokines, interferons and growth factors. In mammals, the JAK family is composed of four members, namely JAK1, JAK2, JAK3 and TYK2. Studies performed on mouse hair follicles have shown that the JAK-STAT pathway is dynamically regulated during the hair cycle. Indeed, the JAK-STAT pathway is activated during the catagen and telogen phases and inhibited early in the anagen phase. Furthermore, topical treatment of telogen with JAK-STAT pathway inhibitors such as tofacitinib (JAK1 / 3>JAK2>TYR2) and ruxolitinib (JAK1 / 2>TYR2>JAK3) has been shown to result in rapid re-entry into early anagen in mice (Harel et al., 2015 Sci. Adv. 1, e1500973). This study also showed that JAK-STAT inhibition in human hair follicles increases hair growth ex vivo. These data suggest that the JAK-STAT signaling pathway may be a novel target for stimulating hair growth.

[0096] The purpose of this example is to evaluate whether mangosteen extract can inhibit the JAK-STAT signaling pathway, as assessed by recombinant human JAK1, JAK2 or JAK3 proteins.

[0097] method Mangosteen extract according to Example 1 is diluted in DMSO and tested at various concentrations (0.1-1000 μg / mL). Positive controls (tofacitinib and ruxolitinib) are also evaluated in this study.

[0098] The test article is incubated with recombinant human JAK1, JAK2 or JAK3 protein together with reaction buffer (Tris / HCl for JAK1, MOPS (3-morpholino-1-propanesulfonic acid) for JAK2 and JAK3), EDTA and the specific peptide substrate. The phosphorylation reaction is then started by adding a mixture of magnesium acetate and radiolabeled ATP (45 μM for JAK1 and JAK2, 10 μM for JAK3). After 40 minutes of incubation at room temperature, the reaction is stopped by adding phosphoric acid. Small molecules including the labeled ATP are eluted by washing four times with phosphoric acid and once with methanol. Finally, the radioactivity of the phosphorylated specific substrate is counted. Compounds are tested in three different experiments, each experiment performed in duplicate.

[0099] Inhibition curves are generated and IC50 values ​​are calculated for each JAK subtype. IC50 (50% inhibitory concentration) is the concentration of a compound that inhibits 50% of the observed effect.

[0100] The IC50 values ​​(in μg / mL) of each recombinant JAK1, JAK2 and JAK3 protein as a function of the test article incubated are shown in Table 1 below.

[0101] [Table 1]

[0102] The results obtained with the reference compounds (ruxolitinib and tofacitinib) are in line with the expected results. Indeed, both compounds strongly inhibit JAK-STAT activity (maximum inhibition of 97-100% and very low IC50 values ​​(in the ng / mL range)). Ruxolitinib shows a higher affinity for JAK1 or JAK2 than for JAK3, and tofacitinib shows a higher affinity for JAK1 or JAK3 than for JAK2. These expected results substantiate the validity of the study.

[0103] Mangosteen extract showed strong inhibition of JAK1 (100% maximal inhibition) with an IC50 value of 6.6 μg / mL, strong inhibition of JAK2 (100% maximal inhibition) with a slightly higher IC50 value of 46.2 μg / mL, and strong inhibition of JAK3 (100% maximal inhibition) with an IC50 value of 1.4 μg / mL.

[0104] This study revealed that mangosteen extract induces inhibition of tyrosine kinase activity (having stronger affinity for JAK1 and JAK3 than JAK2) and has the desired pharmacological activity to promote hair growth.

[0105] Example 12: Confirmation of JAK-STAT pathway inhibitory activity of mangosteen extract at the cellular level The purpose of this example is to confirm the JAK-STAT signal pathway inhibitory activity of mangosteen extract in a cell model, i.e., hair follicle keratinocytes of the outer epithelial sheath of the hair follicle (outer root sheath (ORS) model). In this model, interleukin IL-6 is used to activate the JAK1 / 2-STAT3 pathway through the activation of IL-6 receptor and GP130 receptor, both of which are expressed in the hair follicle epithelial sheath. IL-6 is a cytokine that functions as an inhibitor of the hair growth cycle, and overexpression of IL-6 in a transgenic mouse model results in delayed hair growth. Furthermore, it has been reported that in male pattern baldness, IL-6 is overexpressed in dermal papilla cells under the influence of androgens (Kwack et al., 2012, J Invest Dermatology 132(1)43-9). It has also been reported that IL-6 slows hair follicle growth in humans.

[0106] method The present example is carried out in hair follicle keratinocytes (outer root sheath (ORS) model) derived from the outer epithelial sheath of the hair follicle. The keratinocytes are cultured in 96-well plates using the appropriate medium (CnTa-PR from Cellntec). After 24 hours, the cells are washed with alkaline phosphate buffered saline (PBS) and the medium is changed to CnT-PR-H medium (standard maintenance medium for primary human keratinocytes). After 48 hours, the cells are treated with the test compounds (10 and 30 μg / mL mangosteen extract diluted in DMSO) and the reference compounds (5 μM ruxolitinib diluted in DMSO and 5 μM tofacitinib diluted in DMSO) for 1 hour. The mangosteen extract tested is the extract according to Example 1 of the present invention. Then, IL-6 stimulation treatment is carried out at 100 ng / mL for 15 minutes.

[0107] All conditions are tested on cells from n=3 different donors. Mangosteen extracts are tested in n=3 different experiments, and control conditions for stimulation and control conditions using reference compounds are performed in n=6 experiments. In each experiment, data are obtained in triplicate.

[0108] JAK activity is measured by evaluating the phosphorylation level of STAT3 protein. Statistical analysis is performed by parametric tests after checking normality and equivalence of variance, otherwise non-parametric tests are selected.

[0109] result The results are summarized in Table 2 below.

[0110] [Table 2]

[0111] Treatment of keratinocytes with IL-6 induced a significant increase in the phosphorylation level of STAT3, which reached 19% and was statistically significant (p<0.01). Reference compounds (ruxolitinib and tofacitinib tested at 5 μM) significantly reduced the phosphorylation level of STAT3 by 90% and 59%, respectively (p<0.05 for both compounds). These expected results validate the validity of the study.

[0112] Mangosteen extract significantly and highly significantly inhibited the phosphorylation level of STAT3 induced by IL-6 stimulation at the two concentrations tested.

[0113] Thus, the inventors have demonstrated the usefulness of mangosteen extract in hair regrowth.

[0114] Example 13: Evaluation of mangosteen extracts on versican synthesis and release Anchoring of hair follicles occurs due to a complex organization composed of proteoglycans, matrix proteins (e.g. collagen, etc.) and anchoring structures (e.g. desmosomes, hemidesmosomes, etc.). Proteoglycans are involved in cell adhesion to the extracellular matrix, cell-cell adhesion and cell differentiation. Desmosomes mediate cell-cell adhesion and allow anchoring of the intermediate filament network to the plasma membrane. Research has shown that mutations in hair follicle anchoring proteins, more specifically in desmosomal proteins, result in hypotrichosis in mice as well as in humans (Nagasawa et al., Dermatol Ther (Heidelb), 2016, 6:59-68). Hypotrichosis is a disease of both the skin and scalp and refers to the arrest of all hair growth. It has also been reported that male pattern baldness is associated with miniaturization of hair follicles and ultimately with a loss of anchoring properties.

[0115] Proteoglycans are composed of glycosaminoglycans, which are anionic polysaccharides, bound to basic proteins. Each basic protein preferentially binds to a specific glycosaminoglycan chain. Glycosaminoglycans are classified into chondroitin sulfate, heparan sulfate, keratin sulfate, and dermatan sulfate depending on their chemical structure. It has been shown that the distribution of these proteoglycan components can vary depending on the location and growth state of the hair. Dermal papilla cells contain high levels of basic proteins and a wide variety of glycosaminoglycans. These glycosaminoglycans are known to bind to many biomolecules involved in cell differentiation or proliferation and regulate their functions. These include fibroblast growth factors (FGF), vascular endothelial growth factors (VEGF), sonic hedgehog (Shh), bone morphogenetic proteins (BMPs), wingless integration site (Wnt), hepatocyte growth factor (HGF), etc. All of these factors are well known to be involved in hair follicle morphogenesis. This network also functions as a reservoir of regulatory and growth factors by forming a specific microenvironment for the hair follicle, and is therefore involved in the homeostasis of the hair follicle and the control of hair follicle cell proliferation and differentiation.

[0116] Versican is one of the large proteoglycans of the chondroitin sulfate family. It is produced by fibroblasts, smooth muscle cells and keratinocytes and is involved in the maintenance and firmness of the skin. It is involved in cell adhesion within the extracellular matrix. It plays an important role in cell migration, proliferation and differentiation. It is expressed in the dermal papilla cells of the hair follicle and in the proximal part of the connective tissue sheath, gradually decreasing towards the distal part. The expression of specific genes in the dermal papilla cells is high during the anagen phase of hair and decreases at the beginning of the catagen phase. Its expression is regulated by the β-catenin signaling pathway. This specific expression of versican indicates its importance in the anagen phase of hair.

[0117] The purpose of this example is to evaluate whether mangosteen extract affects the synthesis and release of versican, an anchoring component of hair follicles.

[0118] method The experiments are carried out using human dermal papilla cells derived from hair follicles of three donors. The cells are seeded in 96-well plates and cultured for 24 hours with the necessary supplements. They are then treated with the test product (3 or 10 μg / mL mangosteen extract diluted in DMSO) for 48 hours. The mangosteen extract tested is the extract according to Example 1 of the present invention. At the end of the culture, the supernatant is collected and an ELISA is performed to evaluate the synthesis and release of versican. All experimental conditions are carried out with n=3 donors and n=3 technical replicates. The amount of versican in the supernatant is measured using a specific ELISA kit according to the supplier's instructions (Cusabio).

[0119] result The measurement results of versican concentration in the supernatant are shown in Table 3 below.

[0120] [Table 3]

[0121] Treatment of hair papilla cells with 10 μg / mL of mangosteen extract induces a statistically significant (p<0.01) strong activation of versican synthesis and release. Concentrations of mangosteen extract as low as 3 μg / mL are not sufficient to activate this target. Mangosteen extract inhibits JAK-STAT activity with IC50 values ​​ranging from 1 to 46 μg / mL depending on the subtype, as shown in Example 10. Therefore, it is not illogical that concentrations higher than 3 μg / mL are required to significantly increase proteoglycan synthesis and release, since activation of versican is not related to inhibition of the JAK / STAT pathway.

Claims

1. 1. A hydroalcoholic extract of mangosteen for use in preventing or treating alopecia by promoting hair growth.

2. 2. An extract for use according to claim 1, characterized in that the extract is an extract of the pericarp of the mangosteen fruit.

3. 3. Extract for use according to claim 1 or 2, characterized in that the extract is intended for topical application.

4. 4. The extract for use according to any one of claims 1 to 3, characterized in that the alopecia is selected from the group consisting of androgenetic alopecia, reactive alopecia, postmenopausal alopecia and alopecia areata.

5. Non-therapeutic use of a hydroalcoholic extract of mangosteen to promote hair regrowth.

6. A dermatological composition for use in preventing or treating alopecia by promoting hair growth, comprising at least one hydroalcoholic extract of mangosteen and at least one dermatologically acceptable excipient.

7. Dermatological composition for use according to claim 6, characterized in that said extract is an extract of the pericarp of the mangosteen fruit.

8. Dermatological composition for use according to claim 6 or 7, characterized in that the composition comprises 0.001 to 5% by weight of dry extract, preferably 0.005 to 1% of mangosteen extract, based on the total weight of the composition.

9. Dermatological composition for use according to any one of claims 6 to 8, characterized in that said composition is intended for topical application.

10. Dermatological composition for use according to any one of claims 6 to 9, characterized in that the alopecia is selected from the group consisting of androgenetic alopecia, reactive alopecia, postmenopausal alopecia and alopecia areata.

11. A cosmetic composition for promoting hair growth, in particular for promoting hair regrowth, comprising at least one hydroalcoholic extract of mangosteen and at least one dermatologically acceptable excipient.

12. 12. The cosmetic composition according to claim 11, wherein said hydroalcoholic extract of mangosteen is the only active ingredient for promoting hair growth.

13. 1. Non-therapeutic use of a cosmetic composition comprising at least one hydroalcoholic extract of mangosteen and at least one cosmetically acceptable excipient for promoting hair growth.

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