Compositions for the treatment of hair loss and hair graying

Abstract

The invention is directed to a hair composition for treatment and prevention of alopecia or hair graying. More particularly, the invention, is directed to a composition comprising lutein and phycocyanin. The invention particularly relates to the field of anti-alopecia and anti-graying products.

Description

[0001] COMPOSITIONS FOR THE TREATMENT OF HAIR LOSS AND HAIR GRAYING

[0002] FIELD OF THE INVENTION

[0003] The invention relates to the field of hair care compositions. More specifically, it refers to composition for the treatment and prevention of alopecia and premature loss of hair pigmentation.

[0004] BACKGROUND ART

[0005] The appearance of hair plays an important role in people's overall physical appearance and self-perception. With today's increasing life expectation, the desire to look youthful plays a bigger role than ever. The hair care industry has become aware of this and also more capable to deliver active products that are directed toward meeting this consumer demand.

[0006] Hair loss and premature hair graying are common clinical hair disorders that have a major negative impact on people's mental and psychological well-being. In recent years, with the increasing competition in society, the pressure of people at work, in studying and in life gradually increases, leading to an elevated incidence of hair loss and premature hair graying year by year, with the age of onset getting younger.

[0007] Normally, all humans shed around 50 to 100 of the more than 100,000 hairs on the scalp each day. But problems begin when too many follicles enter rest phase permanently or are damaged or killed. When this happens, the rate of replacement may not keep up, eventually causing the hair to thin out visibly. This excessive hair loss is called alopecia.

[0008] Hair graying in humans is a phenomenon of physiological aging, and is known to be caused by a decrease in melanin pigment due to functional deterioration and depletion of melanocytes or melanocyte stem cells in hair follicles. Melanin pigment is an important factor that determines the color of hair, and melanin is synthesized in and secreted from the melanosomes present in melanocytes, transferred to nearby keratinocytes, fibroblasts, etc., and then moves along with hair growth and maintains the color of the hair. Currently, a simple dyeing procedure is used as a solution to hair graying, but dyeing is a temporary method and re-dyeing is essential due to growth of white hair, and ingredients contained in hair dyes have various side effects, such as irritation to the scalp and hair follicles, or causing allergies or dermatitis.

[0009] The loss of hair in alopecia and premature loss of hair pigmentation are not limited just to head hair but can happen anywhere on the body. Although not usually life-threatening, alopecia and premature loss of hair pigmentation are usually accompanied by serious emotional distress due to issues related to appearance. Accordingly, there is still a need for agents which are adequate for the treatment and the prevention of alopecia and premature loss of hair pigmentation..

[0010] SUMMARY OF THE INVENTION

[0011] The authors of the present invention have developed a comprehensive hair care composition designed to address and prevent alopecia and / or premature loss of hair pigmentation. The disclosed invention presents a unique formulation that combines Lutein and Phycocyanin and that has an unexpected effect in overall hair health.

[0012] Thus, in a first aspect, the present invention relates a composition comprising lutein and phycocyanin for use in the treatment or prevention of alopecia and / or hair graying.

[0013] In a second aspect, the present invention relates to a non-therapeutic cosmetic method for preventing or treating alopecia and / or hair graying comprising the steps of administering a composition comprising lutein and phycocyanin.

[0014] DESCRIPTION OF THE FIGURES

[0015] Figure 1 : Philpott experiment. Human hair follicles were isolated and incubated in vitro with Minoxidyl or the composition for 7 days. Data analysis was performed with Imaged software.

[0016] Figure 2: In vivo hair growth study measured by Phototricogram. Phototrichogram is carried out as recommended by TrichoScan® supplier.

[0017] Figure 3: In vivo hair density study measured by Phototricogram. Phototrichogram is carried out as recommended by TrichoScan® supplier.

[0018] Figure 4: In vivo hair graying study measured by colorimeter. The instrument evaluates the colour according to a standard color space defined by the International Lighting Commission (CIE).

[0019] Figure 5: In vivo scalp moisturization by conductance measurement. The method refers to conductance measurement. The hydration state is presented in the unit of microSiemens (pS). An increase of this index reflects an increase of the moisturisation.

[0020] DETAILED DESCRIPTION OF THE INVENTION The authors of the present invention have developed a comprehensive hair care composition designed to address and prevent alopecia and / or premature hair graying. The disclosed invention presents a unique formulation that combines lutein and phycocyanin to overall hair health.

[0021] The invention is defined by the following aspects:

[0022] In a first aspect, the present invention relates to a composition comprising lutein and phycocyanin for use in the treatment or prevention of alopecia or premature hair graying.

[0023] “Combination” or “composition”, as used herein, indicates that the lutein and the phycocyanin can be formulated in the same formulation or they can also be formulated in different ones.

[0024] As used herein, the terms “comprising”, “including”, “having” and grammatical variants thereof are to be taken as specifying the stated features, steps or components but do not preclude the addition of one or more additional features, steps, components or groups thereof.

[0025] The term “lutein”, as used herein, refers to the compound p, e-Carotene-3, 3'-diol or (1 / ?, 4 / ?)-4-{( 1 E,3E,5E,7E,9E, 11 E, 13E, 15E, 17E)-18-[(4F?)-4-Hydroxy-2,6,6- trimethylcyclohex-1-en-1-yl]-3,7,12,16-tetramethyloctadeca-1 ,3,5,7,9,11 ,13,15,17- nonaen-1-yl}-3,5,5-trimethylcyclohex-2-en-1-ol, and defined under the CAS number 127- 40-2.

[0026] As used herein, lutein is in the form of free xanthophylls, xanthophyll esters or other chemical forms of lutein. Lutein may be obtained or isolated by any method recognized by those skilled in the art. For example, lutein may be obtained by extraction from microalgae and, in particular, from Chlorella vulgaris, from marigolds or other xanthophylls-rich sources, chemical synthesis, fermentation or other biotechnology- derived and enriched xanthophyll sources.

[0027] As used herein, the term “phycocyanin” refers to the compound defined under the CAS number 11016-15-2. Phycocyanin is a pigment-protein complex from the light-harvesting phycobiliprotein family, along with allophycocyanin and phycoerythrin. Phycocyanins are found in some organisms such as cyanobacteria. It is an accessory pigment to chlorophyll.

[0028] The term “treatment”, as used herein, refers to any type of therapy, which is aimed at terminating, preventing, ameliorating or reducing the susceptibility to a clinical condition as described herein. In a preferred embodiment, the term treatment relates to prophylactic treatment (i.e. a therapy to reduce the susceptibility to a clinical condition), of a disorder or a condition as defined herein. Thus, “treatment,” “treating,” and their equivalent terms refer to obtaining a desired pharmacologic or physiologic effect, covering any treatment of a pathological condition or disorder in a mammal, including a human. The effect may be prophylactic in terms of completely or partially preventing a disorder or symptom thereof and / or may be therapeutic in terms of a partial or complete cure for a disorder and / or adverse effect attributable to the disorder. That is, “treatment” includes (1) preventing the disorder from occurring or recurring in a subject, (2) inhibiting the disorder, such as arresting its development, (3) stopping or terminating the disorder or, at least, symptoms associated therewith, so that the host no longer suffers from the disorder or its symptoms, such as causing regression of the disorder or its symptoms, for example, by restoring or repairing a lost, missing or defective function, or stimulating an inefficient process, or (4) relieving, alleviating, or ameliorating the disorder, or symptoms associated therewith, where ameliorating is used in a broad sense to refer to at least a reduction in the magnitude of a parameter.

[0029] The term “prevention”, “preventing” or “prevent”, as used herein, relates to the administration of a combination according to the invention orof a medicament comprising said combination to a subject who has not been diagnosed as possibly having a alopecia as an example, but who would normally be expected to develop said disease or be at increased risk for said disease. The prevention intends to avoid the appearance of said disease. The prevention may be complete (e.g. the total absence of a disease). The prevention may also be partial, such that for example the occurrence of a disease in a subject is less than that which would have occurred without the administration of the composition of the present invention. Prevention also refers to reduced susceptibility to a clinical condition.

[0030] As used herein, “alopecia” refers to the involuntary complete or partial hair loss from the head or body of an individual.

[0031] In a particular embodiment, the composition for use according to first aspect of the invention, the alopecia is selected from the group consisting of: (a) androgenic alopecia, (b) traction alopecia, (c) anagen effluvium, (d) telogen effluvium, (e) alopecia areata, (f) alopecia totalis, (g) alopecia universalis, (h) alopecia barbae, (i) alopecia mucinosa, (j) alopecia neoplastica, (k) cicatricial alopecia, and (I) scarring alopecia. As used herein, "androgenic alopecia", also known as male-pattern hair loss, is hair loss that occurs due to an underlying susceptibility of hair follicles to shrinkage due to the influence of androgenic hormones.

[0032] The term "traction alopecia", as used herein, means a form of alopecia (hair loss or hair shedding) associated with mechanical forces that pull the hair such as hair brushing hair combing, fiat ironing, wearing of extensions, hair braiding, and ponytail style hair. Under this definition, although chronic traction on the hair can lead to traction alopecia, the mechanical forces that pull the hair do not necessarily need to be chronic to lead to hair loss or excessive shedding.

[0033] As used herein, “anagen effluvium” refers to a hair loss due to chemicals or radiation, such as chemotherapy or radiation treatment for cancer.

[0034] The term "telogen effluvium", as used herein, refers to a scalp disorder characterized by the thinning or shedding of hair resulting from the early entry of hair in the telogen phase (the resting phase of the hair follicle)

[0035] As used herein, "alopecia areata", also known as spot baldness, is an autoimmune disease in which hair is lost from some or all areas of the body due to the body's failure to recognize its own cells and subsequent destruction of its own tissue.

[0036] The term "alopecia totalis", as used herein, refers to the loss of all head hair.

[0037] As used herein, "alopecia universalis" refers to a condition characterized by the complete loss of hair on the scalp and body. It is an advanced form of alopecia areata, a condition that causes round patches of hair loss.

[0038] The term “alopecia barbae”, as used herein, refers to a condition characterized by the loss of some or all of their beard.

[0039] As used herein, “alopecia mucinosa”, also known as follicular mucinosis, refers to a skin disorder that generally presents as erythematous plaques or flat patches without hair primarily on the scalp, neck, and face. It is characterized by the appearance of mucin around hair follicles as seen under the microscope. This condition results from the deposition of a jelly-like substance (called mucin) in and around the hair follicles.

[0040] The term “alopecia neoplastica”, as used herein, refers to a rare form of hair loss that is associated with malignant tumors or metastases. It typically presents as solitary or multiple smooth patches or plaques of scarring hair loss, appearing anywhere on the scalp. The hairless areas may be skin-colored or pinkish.

[0041] As used herein, "cicatricial alopecia" also called scarring alopecia, refers to a group of rare disorders that destroy hair follicles. The follicles are replaced with scar tissue, causing permanent hair loss. As used herein, "lichen planopilaris" is a type of scarring hair loss that occurs when a relatively common skin disease, known as lichen planus, affects areas of the skin with hair. Lichen planopilaris destroys the hair follicle replacing it with scarring.

[0042] As used herein, "ring alopecia" is a ring or band of alopecia encircling or partially encircling the head; it may extend along the posterior occipital area, around the temporal portion of the scalp above the ears or onto the forehead.

[0043] The terms “loss of hair pigmentation”, “canities” and "hair graying” refer to a consequence of melanogenesis reduction during the anagen phase of the hair cycle and is determined by precise genetic control and by a physiological slowing down protein synthesis.. In some embodiments, hair graying is cause by specific pathological conditions (albinism, piebaldism, phenylketonuria, vitiligo, etc.), which may have to be considered in very early (before the second decade) and extremely sudden cases of white hair.

[0044] As used herein, the term “premature loss of hair pigmentation” also known as premature graying hair (PGH) is defined as graying of hair before the age of 20 years in Caucasians and before 30 years in African American population.

[0045] The second aspect of the present invention relates to a non-therapeutic cosmetic method for preventing and / or treating alopecia or premature hair graying comprising the steps of administering a composition comprising lutein and phycocyanin.

[0046] The term “non-therapeutic cosmetic method”, as used herein, relates to a method used to prevent or treat alopecia in a subject. Cosmetic compositions used in the cosmetic method of the invention include, but are not limited to, skin-care creams, serums, lotions, powders, lipsticks, eye and facial makeup, towelettes, gels, deodorants, hand sanitizer, baby products, bath oils, bubble baths, bath salts, butters and many other types of products.

[0047] By “contacting” or "administering" or "application" it is meant that the composition of the invention is delivered to the subject in such a way that it can achieve the desired purpose. In a particular embodiment, the lutein that forms part of the compositions for use according to the first and second aspects of the invention is encapsulated into liposomes.

[0048] In a particular embodiment, the phycocyanin that forms part of the compositions for use according to the first and second aspects of the invention is encapsulated into liposomes.

[0049] In a particular embodiment, both the lutein and the phycocyanin that forms part of the compositions for use according to the first and second aspects of the invention are encapsulated into liposomes. It will be understood that the compositions for use according to the invention may contain a single type of liposomes which include both lutein and phycocyanin or two types of liposomes containing each lutein and phycocyanin.

[0050] As used herein, the term “liposome” is used to define a variety of single and multilamellar lipid vehicles formed by the generation of enclosed lipid bilayers or aggregates. Liposomes may be characterized as having vesicular structures with a bilayer membrane, generally comprising a phospholipid, and an inner medium that generally comprises an aqueous composition. Liposomes provided herein include, but are not limited to, unilamellar liposomes, multilamellar liposomes and multivesicular liposomes. Liposomes provided herein may be positively charged, negatively charged or neutrally charged.

[0051] In certain embodiments, a liposome may comprise one or more of the following lipids: cholesterol, phosphatidylcholine, hydrogenated soy phosphatidylcholine, dipalmitoyl phosphatidylcholine, phosphatidylglycerol, phosphatidic acid, phosphatidylethanolamine, 2-distearyl-sn-glycero-3-phosphoethanolamine, phosphatidylserine, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-triethylammonium (DOTMA), and 1 ,2-dioleoyl-3-trimethylammoniopropane (DOTAP).

[0052] The lutein-containing liposomes, the phycocyanin-containing liposomes or the lutein and phycocyanin-containing liposomes that can be included in the compositions according to the present embodiments can be made by different methods, as would be known to one of ordinary skill in the art.

[0053] The size of the liposomes forming part of the compositions to be used in the methods according to the present invention varies depending on the method of synthesis. However, this parameter is not particularly limiting of the invention. Liposomes in the present embodiments can be a variety of sizes. In certain embodiments, the liposomes are small, e.g., less than about 500 nm, less than about 400 nm, less than about 300 nm, less than about 200 nm, about 100 nm in external diameter.

[0054] In another particular embodiment, the composition for use according to the invention is a dispersion containing liposomes comprising lutein, liposomes comprising phycocyanin or liposomes comprising lutein and phycocyanin which are dispersed in an aqueous solution. In this case, the compositions for use in the methods according to the invention will contain an aqueous phase.

[0055] The aqueous phase of a composition according to the invention comprises water and optionally a water-soluble solvent. In the present invention, the term "water-soluble solvent" denotes a compound that is liquid at room temperature and water-miscible (miscibility with water of greater than 50 percent by weight at 25 degrees centigrade and atmospheric pressure). The water-soluble solvents that may be used in the composition of the invention may also be volatile.

[0056] Among the water-soluble solvents that may be used in the composition in accordance with the invention, mention may be made especially of lower monoalcohols containing from 2 to 8 carbon atoms, such as ethanol and isopropanol, glycols containing from 2 to 8 carbon atoms, such as ethylene glycol, propylene glycol, 1 ,3 -butylene glycol and dipropylene glycol, C3 and C4 ketones and C2-C4 aldehydes.

[0057] The aqueous phase (water and optionally the water-miscible solvent) may be present in the composition in a content ranging from 20 percent to 95 percent, better still from 30 percent to 80 percent by weight by weight relative to the total weight of the said composition.

[0058] In one embodiment, the concentration of lutein in the compositions is of between 0.001 mM to 0.5 mM. In some embodiment, the concentration of lutein is between 0.050 mM to 0.45 mM or 0.100 mM to 0.4 mM or 0.150 mM to 0.35 mM or 0.200 mM to 0.3 mM.

[0059] In another particular embodiment, the concentration of phycocyanin is between 0.01 mg / mL to 2 mg / mL. In some embodiment, the concentration of phycocyanin is between 0.1 mg / mL to 1.9 mg / mL or 0.2 mg / mL to 1.8 mg / mL or 0.3 mg / mL to 1.7 mg / mL or 0.4 mg / mL to 1.6 mg / mL or 0.5 mg / mL to 1.5 mg / mL or 0.6 mg / mL to 1.4 mg / mL or 0.7 mg / mL to 1.3 mg / mL or 0.8 mg / mL to 1.2 mg / mL or 0.9 mg / mL to 1.1 mg / mL.

[0060] In an embodiment, the lutein is provided as a microalgae extract. The term “microalgae extract” refers to a product obtained from microalgae, for example, by subjecting a microalgae culture to specific treatments. The components present in a microalgal extract will vary depending on the microalgae and the treatments applied thereon. The term “algae”, as used herein, relates to a large and diverse group of simple, typically autotrophic organisms, ranging from unicellular to multicellular forms, including both macroalgae and microalgae, i.e., microscopic algae, typically found in freshwater and marine systems. In the context of the invention, the alga is a microalga, particularly a lutein and / or phycocyanin producing microalga.

[0061] The microalgal extract can include components other than glycolipid and phospholipid, for example, neutral lipids (e.g., triacylglycerol (TAG)), fatty acids, sterols, pigments, nitrogenous compounds, vitamins, carotenoids, phycocyanin, and the like. When present, fatty acids can include, for example, polyunsaturated fatty acids (PLIFA), monounsaturated fatty acids (MLIFA), saturated fatty acids, free fatty acids (FFA), fatty acid methyl esters (FAME), omega-6 fatty acids, omega 3-fatty acids, and mixtures thereof. For example, the microalgal extract can include at least about 1 , 2, 3, 4, 5, 10, 15, 20, 25, or 30 wt percent fatty acids, based on the total weight of the extract and / or up to about 55, 50, 45, 40, 35, 30, 25, or 20 wt percent fatty acids, based on the total weight of the extract.

[0062] In a particular embodiment, the microalgae extract comprises between 0.1 mM and 5 mM of lutein. In some embodiment, the concentration of lutein in the microalgae extract is between 0.2 mM and 4.8 mM or 0.4 mM and 4.6 mM or 0.6 mM and 4.4 mM or 0.8 mM and 4.2 mM or 1 mM and 4 mM or 1.2 mM and 3.8 mM or 1.4 mM and 3.4 mM or 1 .8 mM and 3.2 mM or 2 mM and 3 mM or 2.2 mM and 2.8 mM or 2.4 mM and 2.6 mM.

[0063] In another particular embodiment, the microalgae extract from where the lutein is provided belongs to the genus Chlorella. The Chlorella genus includes, without limitation, Chlorella autotrophica, Chlorella minutissima, Chlorella pyrenoidosa, Chlorella variabilis and Chlorella vulgaris. In a more particular embodiment, the microalgae extract from where the lutein is provided is a Chlorella vulgaris extract.

[0064] In another embodiment, the phycocyanin is provided as a microalgae extract. The term “microalgae extract” has been defined above in the context of the extracts containing lutein and applies equally to the extracts containing phycocyanin.

[0065] In a more particular embodiment, the microalgae extract content comprises between 0.05 mg / mL and 2 mg / mL of phycocyanin. In some embodiment, the concentration of phycocyanin in the microalgae extract is between 0.1 mg / mL to 1.9 mg / mL or 0.2 mg / mL to 1.8 mg / mL or 0.3 mg / mL to 1.7 mg / mL or 0.4 mg / mL to 1.6 mg / mL or 0.5 mg / mL to 1 .5 mg / mL or 0.6 mg / mL to 1 .4 mg / mL or 0.7 mg / mL to 1 .3 mg / mL or 0.8 mg / mL to 1 .2 mg / mL or 0.9 mg / mL to 1.1 mg / mL.

[0066] In one embodiment, the microalgae extract used as a source of lutein or phycocyanin is an extract obtained using a polar solvent.

[0067] As used herein, the term “polar solvent” refers to a solvent that comprises dipole moments. For example, a polar solvent can be miscible with water and polar solvents. For example, a polar solvent can comprise chemical species in which the distribution of electrons between covalently bonded atoms is not even. The polarity of solvents can be assessed by measuring any parameter known to those of skill in the art, including dielectric constant, polarity index, and dipole moment. The polar extracts of the invention can comprise any percentage of polar solvent including, but not limited to, for example 1-10 percent polar solvent, 10- 20 percent polar solvent, 20-30 percent polar solvent, 30- 40 percent polar solvent, 40-50 percent polar solvent, 50- 60 percent polar solvent, 70- 80 percent polar solvent, 80-90 percent polar solvent and 90-100 percent polar solvent. Examples of polar solvents include but are not limited to ethyl alcohol (ethanol), butyl alcohol (butanol), methanol, water, acetic acid, tetra hydrofuran, ethylene glycol, propylene glycol, butylene glycol, pentylene glycol, hexylene glycol, pentanol, hexanol, butanediol, pentanediol, hexanediol, octanediol, glycerl, N,N-dimethylformamide, dichloromethane, acetonitrile, dimethylformamide, dimethyl sulfoxide, acetone, or n- propanol.

[0068] In one embodiment, the microalgae extract used as a source of lutein or phycocyanin is an extract obtained using a non-polar solvent.

[0069] As used herein, the term "non-polar solvent" can refer to a solvent comprising molecules that do not have an overall dipole. For example, the solvent comprises molecules comprising bonds between atoms with similar electronegativities (e.g., a carbonhydrogen bond). For example, the nonpolar molecule comprises equal sharing of electrons between atoms or the arrangement of polar bonds leads to overall no net molecular dipole moment. The non-polar extracts of the invention can comprise any percentage of non-polar solvent, including but not limited to, for example, 1-10 percent non-polar solvent, 10-20 percent non-polar solvent, 20-30 percent non-polar solvent, 30- 40 percent non-polar solvent, 40-50 percent non-polar solvent, 50-60 percent non-polar solvent, 70-80 percent non-polar solvent, 80-90 percent non-polar solvent, and 90-100 percent non-polar solvent. Examples of non-polar solvents include but are not limited to supercritical carbon dioxide (scCO2), isooctane, hexane, pentane, benzene, chloroform, diethyl ether, ethyl acetate, hydrocarbons, cyclohexane, toluene, or 1 ,4- dioxane.

[0070] In some embodiments, the lutein is provided as a Chlorella vulgaris extract obtained using alcohol.

[0071] In some embodiments, the phycocyanin is provided as a Spirulina extract obtained in a polar solvent which is any of the those which appear in Table 1 in the column “Solvent” (pages 4-6) of Pez Jaeschke, D. et al. (Food Res. Int. , 2021 , 143, 110314).

[0072] In a particular embodiment the microalgae extract from where the phycocyanin is an Arthrospira sp. extract. Arthrospira genus includes, but is not limited to the species Arthrospira maxima and Arthrospira platensis.

[0073] In another embodiment, the composition is in the form of a solid, a powder, a lotion, a cream, a gel, a paste, an oil, a serum, a mask, a foam, a mousse, a stick, an aerosol and similar which may be formulated according to conventional methods that use suitable excipients, such as, for example, emulsifiers, surfactants, thickening agents, coloring agents and combination of two or more thereof.

[0074] In a more particular embodiment, the composition is applied topically. “Topical application" means to apply or spread the formulation, composition or product of the present embodiments onto the surface of the keratinous tissue. In some embodiments, a formulation as described is in a form of a cream, an ointment, a paste, a gel, a lotion, a milk, a suspension, a solution, an aerosol, a spray, a foam, a serum, or a mousse.

[0075] The formulation can be water-based, or emulsion-based (including water-in-oil, oil-in- water, water-in-oil-in-water and oil-in-water-in-oil emulsions) or silicon-based.

[0076] Ointments are semisolid preparations, typically based on petrolatum or petroleum derivatives. The specific ointment base to be used is one that provides for optimum delivery for the active agent chosen for a given formulation, and, preferably, provides for other desired characteristics as well (e.g., emolliency). As with other carriers or vehicles, an ointment base should be inert, stable, nonirritating and nonsensitizing. As explained in Remington: The Science and Practice of Pharmacy, 19th Ed., Easton, Pa.: Mack Publishing Co. (1995), pp. 1399-1404, ointment bases may be grouped in four classes: oleaginous bases; emulsifiable bases; emulsion bases; and water-soluble bases. Oleaginous ointment bases include, for example, vegetable oils, fats obtained from animals, and semisolid hydrocarbons obtained from petroleum.

[0077] Emulsifiable ointment bases, also known as absorbent ointment bases, contain little or no water and include, for example, hydroxystearin sulfate, anhydrous lanolin and hydrophilic petrolatum. Emulsion ointment bases are either water-in-oil (W / O) emulsions or oil-in-water (O / W) emulsions, and include, for example, cetyl alcohol, glyceryl monostearate, lanolin and stearic acid. Preferred water-soluble ointment bases are prepared from polyethylene glycols of varying molecular weight.

[0078] Lotions are preparations that are to be applied to the skin surface without friction. Lotions are typically liquid or semiliquid preparations in which solid particles, including the sunscreens- containing capsules, are present in a water or alcohol base. Lotions are typically preferred for covering / protecting large body areas, due to the ease of applying a more fluid composition. Lotions are typically suspensions of solids, and oftentimes comprise a liquid oily emulsion of the oil-in-water type. It is generally necessary that the insoluble matter in a lotion be finely divided. Lotions typically contain suspending agents to produce better dispersions as well as compounds useful for localizing and holding the active agent in contact with the skin, such as methylcellulose, sodium carboxy methylcellulose, and the like.

[0079] Creams are viscous liquids or semisolid emulsions, either oil-in-water or water-in-oil. Cream bases are typically water-washable, and contain an oil phase, an emulsifier and an aqueous phase. The oil phase, also called the "internal" phase, is generally comprised of petrolatum and / or a fatty alcohol such as cetyl or stearyl alcohol and / or natural oil substances and / or plant extracts. The aqueous phase typically, although not necessarily, exceeds the oil phase in volume, and generally contains a humectant. The emulsifier in a cream formulation is generally a nonionic, anionic, cationic or amphoteric surfactant. Reference may be made to Remington: The Science and Practice of Pharmacy, supra, for further information.

[0080] Pastes are semisolid dosage forms in which the bioactive agent is suspended in a suitable base. Depending on the nature of the base, pastes are divided between fatty pastes or those made from a single-phase aqueous gels. The base in a fatty paste is generally petrolatum, hydrophilic petrolatum and the like. The pastes made from singlephase aqueous gels generally incorporate carboxymethylcellulose or the like as a base. Additional reference may be made to Remington: The Science and Practice of Pharmacy, for further information. Gel formulations are semisolid, suspension-type systems. Single-phase gels contain organic macromolecules distributed substantially uniformly throughout the carrier liquid, which is typically aqueous, but also, preferably, contain an alcohol and, optionally, an oil. Preferred organic macromolecules, i.e., gelling agents, are crosslinked acrylic acid polymers such as the family of carbomer polymers, e.g., carboxypolyalkylenes that may be obtained commercially under the trademark Carbopol™. Other types of preferred polymers in this context are hydrophilic polymers such as polyethylene oxides, polyoxyethylene-polyoxypropylene copolymers and polyvinylalcohol; cellulosic polymers such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, and methyl cellulose; gums such as tragacanth and xanthan gum; sodium alginate; and gelatin. In order to prepare a uniform gel, dispersing agents such as alcohol or glycerin can be added, or the gelling agent can be dispersed by trituration, mechanical mixing or stirring, or combinations thereof.

[0081] Sprays generally provide the active agent in an aqueous and / or alcoholic solution which can be misted onto the skin for delivery. Such sprays include those formulated to provide for concentration of the active agent solution at the site of administration following delivery, e.g., the spray solution can be primarily composed of alcohol or other like volatile liquid in which the active agent can be dissolved. Upon delivery to the skin, the carrier evaporates, leaving concentrated active agent at the site of administration.

[0082] Foam compositions are typically formulated in a single or multiple phase liquid form and housed in a suitable container, optionally together with a propellant which facilitates the expulsion of the composition from the container, thus transforming it into a foam upon application. Other foam forming techniques include, for example the "Bag-in-a-can" formulation technique. Compositions thus formulated typically contain a low-boiling hydrocarbon, e.g., isopropane. Application and agitation of such a composition at the body temperature cause the isopropane to vaporize and generate the foam, in a manner similar to a pressurized aerosol foaming system. Foams can be water-based or hydroalcoholic, but are typically formulated with high alcohol content which, upon application to the skin of a user, quickly evaporates, driving the active ingredient through the upper skin layers to the site of treatment.

[0083] Serum composition refers to an aqueous solution which is formulated for topical applications. It is a concentrated liquid treatment designed to deliver active ingredients. Serum has a very fine texture. It penetrates fast into the skin and can be applied to the skin in very small quantities. Serums are characterized by having a lightweight texture and are formulated with potent ingredients to target specific skin concerns such as wrinkles, discoloration, and uneven texture. Some non-limiting representative examples of additives and / or agents that may be added to the compositions for use in the methods according to the present invention include humectants, antioxidants, solvents, odor absorbers, perfumes, deodorants, antiperspirants, sunscreen agents (e.g., UV blocking agents, UV filters), sunless tanning agents, hair conditioning agents, pH adjusting agents, chelating agents, preservatives, emulsifiers, occlusive agents, emollients, thickeners, solubilizing agents, penetration enhancers, anti-irritants, colorants or coloring agents (pigments, nacres, water-soluble dyestuffs), propellants, surfactants, dispersants, fillers and bactericides. Representative examples of humectants include, without limitation, guanidine, glycolic acid and glycolate salts (e.g. ammonium slat and quaternary alkyl ammonium salt), aloe vera in any of its variety of forms (e.g., aloe vera gel), allantoin, urazole, polyhydroxy alcohols such as sorbitol, glycerol, hexanetriol, propyleneglycol, butylene glycol, hexylene glycol and the like, polyethylene glycols, sugars and starches, sugar and starch derivatives (e.g., alkoxylated glucose), hyaluronic acid, lactamide monoethanolamine, acetamide monoethanolamine and any combination thereof.

[0084] The invention is described below by means of the following examples which are to be construed as merely illustrative and not limitative of the scope of the invention.

[0085] EXAMPLES

[0086] Materials

[0087] Preparation, purification and encapsulation of a Chlorella vulgaris extract

[0088] Chlorella vulgaris dry biomass is extracted with alcohol and filtered. The resulting organic extract was purified by means of a strong basic anionic resin in order to selectively remove chlorophylls and esterified fatty acids. The resin is separated from the extract by filtration and the purified extract was encapsulated in liposomes by the thin-layer hydration method.

[0089] Preparation of a microalgae extract containing phycocyanin

[0090] Spirulina cells are commonly disrupted by freezing and thawing cycles. The extraction may be performed at moderate temperatures (up to 50°C), neutral pH values (6-8) and preventing light contact. Preparation of the combination

[0091] The composition contains lutein from a green seaweed from the species Chlorella vulgaris and phycocyanin from a cyanobacteria from the genus Arthrospira.

[0092] A Chlorella vulgaris extract containing lutein was encapsulated. Afterwards, a phycocyanin solution in a buffer at pH 7 was mixed with the encapsulated extract and the mixture was encapsulated in the liposomes by high pressure homogenization. The resulting solution was filtered with a 0.7 pm filter to obtain the desired combination.

[0093] Results

[0094] Example 1 - Philpott Study:

[0095] Human hair follicles were isolated by manual extraction from the human nape of several volunteers. Hair follicles in the anagen phase were selected through morphologic discrimination and maintained in growth medium with Fungizone, penicillin and streptomycin supplements as previously described [Philpott et al. 1989], Follicles were maintained free-floating in individual wells of 24-well plates, at 37°C in an atmosphere of 5 % CO2 / 95 % air. Basal images (Oh) were taken immediately after isolation using a stereomicroscope. Then, Minoxidil or composition containing lutein and phycocyanin at 0.5 % concentration, or positive control (EGF, Epidermal Growth Factor) were added to each well and plates were incubated at 37oC for 7 days. Images were taken after 7 days of treatment, under identical conditions. Finally, data analysis was performed using Imaged software.

[0096] Results showed that treatment with Minoxildyl stimulated hair growth, but the change was not statistically significant. On the other hand, treatment with the new active significantly stimulated hair growth on human hair follicles for 7 days, compared to the non-treated control as shown in Fig. 1.

[0097] Example 2 - Efficacy from In Vivo Studies:

[0098] Fifty adult subjects enrolled to assess the invention's efficacy in reducing hair loss and hair graying compared to a placebo after 112 days.

[0099] Evaluation methods include phototrichogram, scalp skin moisturization, and hair color intensity measurements. Results shown in Fig 2, Fig. 3, Fig. 4 and Fig. 5.

[0100] Phototricogram. Phototrichogram is carried out as recommended by TrichoScan® supplier. Pictures are taken 48 hours after shaving. Thephototrichogram procedure involves: i) clipping of a scalp area two fingers width away from the parting on the receding hairline of the fronto-temporal regions or on the vertex. A template (0 1 .8 cm) is used; ii) removal of the clipped hair with sticky tape; iii) application of dye on the clipped scalp region; iv) removal of dye remnants by means of a swab and an alcohol solution; v) photograph taking. Anagen and hair density are automatically calculated by TrichoScan® software.

[0101] Scalp skin moisturization (DermaLab-Pin probe® - CORTEX TECHNOLOGY, Aalborg, Denmark). The method refers to conductance measurement. The hydration state is presented in the unit of micro-Siemens (pS). An increase of this index reflects an increase of the moisturization.

[0102] Evaluation of hair color intensity (20 subjects with salt and pepper hair): The measurement of hair colour intensity is done by means of a colorimeter CM-700D (Konica Minolta). The instrument evaluates the colour according to a standard color space defined by the International Lighting Commission (CIE).

Claims

CLAIMS1. A composition comprising lutein and phycocyanin for use in the treatment or prevention of alopecia and / or premature loss of hair pigmentation.

2. The composition for use according to claim 1, wherein the alopecia is selected from the group consisting of: (a) androgenic alopecia, (b) traction alopecia, (c) anagen effluvium, (d) telogen effluvium, (e) alopecia areata, (f) alopecia totalis, (g) alopecia universalis, (h) alopecia barbae, (i) alopecia mucinosa, (j) alopecia neoplastica, (k) cicatricial alopecia, and (I) scarring alopecia.

3. A non-therapeutic cosmetic method for preventing or treating alopecia and / or loss of hair pigmentation comprising the steps of administering a composition comprising lutein and phycocyanin.

4. The composition for use according to claims 1 or 2 or the method according to claim 3, wherein the lutein and phycocyanin are encapsulated into liposomes.

5. The composition for use or the method according to claim 4 wherein the liposomes comprising lutein and phycocyanin are dispersed in an aqueous solution.

6. The composition for use according to any one of claims 1 to 5 or the method according to any one of claims 3 to 5, wherein the lutein is at a concentration of 0.001 mM to 0.5 mM.

7. The composition for use according to any one of claims 1 to 6 or the method according to any one of claims 3 to 6, wherein the phycocyanin is at a concentration of 0.01 mg / mL to 2 mg / mL.

8. The composition for use according to any one of claims 1 to 7 or the method according to any one of claims 3 to 7, wherein the lutein is provided as a microalgae extract.

9. The composition for use or the method according to claim 8 wherein the microalgae extract comprises between 0.1 mM and 5 mM of lutein10. The composition for use or the method according to claims 8 or 9 wherein the microalgae extract is a Chlorella vulgaris extract.

11. The composition for use according to any one of claims 1 to 10 or the method according to any one of claims 3 to 10, wherein the phycocyanin is provided as a microalgae extract.

12. The composition for use or the method according to claim 11 wherein the microalgae extract content comprises between 0.05 mg / mL and 2 mg / mL of phycocyanin.

13. The composition for use or the method according to claims 11 or 12 wherein the microalgae extract is an extract obtained using a polar solvent.

14. The composition for use or the method according to claims 11 to 13 wherein the microalgae extract is an Arthrospira sp. extract.

15. The composition for use according to any one of claims 1 to 14 or the method according to any one of claims 3 to 14, wherein the composition is in the form of a solid, a powder, a lotion, a cream, a gel, a paste, an oil, a serum, a mask, a foam, a mousse, a stick or an aerosol.

16. The composition for use according to any one of claims 1 to 15 or the method according to any one of claims 3 to 15, wherein the composition is applied topically.

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