KR20190016428A - Composition for Preventing Hair Loss or Stimulating Hair Growth Comprising Phosphodiesterase 3 inhibitor - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or promoting hair growth, containing a phosphodiesterase (PDE) 3 inhibitor as an active ingredient and, more specifically, to a composition for preventing hair loss or promoting hair growth, which inhibits the action of PDE 3, thereby exhibiting excellent hair loss prevention or hair growth promotion effects. A typical PDE 3 inhibitor, cilostazol, is already being used as an antiplatelet and the safety thereof has been proven. According to the present invention, it has been confirmed that a PDE 3 inhibitor including the cilostazol promotes proliferation of human-derived hair papilla cells (hDPCs) and promotes hair growth and thus excellent effects on hair loss or hair growth are secured. Accordingly, the composition of the present invention is expected to be useful in developing a therapeutic agent for preventing hair loss and promoting hair growth, or a scalp applying cosmetic material.

Description

TECHNICAL FIELD The present invention relates to a composition for preventing hair loss or promoting hair growth comprising a phosphodiesterase 3 inhibitor,

The present invention relates to a composition for preventing hair loss or promoting hair growth comprising a phosphodiesterase 3 (PDE 3) inhibitor as an active ingredient.

The hair of the human body grows, maintains and disappears according to the growth cycle through three stages of anagen, catagen, and telogen. It is known that in the case of adults, the hair grows about 0.3mm per day, grows about 1cm per month, and lasts for about 5 to 8 years. In general, the hair is removed through a catagen and an average telogen of about 3 weeks after the growth, and the reason why the length of the human hair varies according to the site is that the growth characteristic of the hair follicle This is due to the fact that the duration of

The hair is formed in hair follicles, and the hair follicle is a very complicated organs composed of the hair follicles, the hair follicle, the hair follicle, the hair follicle cell, and the hair dermal papilla cell (hDPC) , Which plays a key role in the formation of hair follicles. If the hair follicle cell is absent or does not function properly, the proper interaction between the papillary cell and the epidermal cell can not be established, so that the hair follicle can not be formed.

Hair loss refers to a state of hair with little or no hair on the part where hair should normally exist. It is caused by hair loss, genetic cause, male hormone action as well as endocrine diseases, malnutrition, drug use, birth, fever, Physical and mental stress, and so on, are known to cause hair loss symptoms. Recently, the hair loss population of the female is increasing due to the increase of the stress due to the change of the eating habits, the social environment and the like as well as the male hair loss, and the population suffering from the abnormal symptoms of the scalp and the hair is increasing, It is getting lower.

Research on anti-hair-loss agents and hair growth agents has been accelerated at home and abroad, and various products have been developed. However, excellent anti-hair-loss agents with excellent efficacy and safety to the human body have not been developed yet. Minoxidil preparations Despite the fact that herbal medicine extracts, mainly in Japan, are being developed as hair restorers and hair growth promoters, it is still too early to seek hair loss prevention function and hair growth in terms of their efficacy.

Currently, only 5a reductase inhibitors such as finasteride and dutasteride are effective. However, there are many individual differences according to the treatment effect. In the case of existing hair loss treatment drugs approved by US FDA, , It is difficult to solve the problem of female alopecia of women and there is a possibility of causing side effects rather to women. Therefore, it is very necessary to study new anti-hair-loss agents and hair growth agents that can demonstrate safety and efficacy in both men and women.

On the other hand, the phosphodiesterase 3 (PDE 3) inhibitor inhibits the action of the 3 subtype enzyme in the 10 or more phosphodiesterase subtypes, resulting in the formation of intracellular secondary messenger cyclic adenosine monophosphate (cyclic adenosine monophosphate (cAMP)) to affect various gene transcription, protein expression control and various physiological processes mediated by cAMP, and to treat diseases such as acute heart failure, cardiac shock, intermittent claudication Is currently used as a target for inhibition of various pharmacological mechanisms. However, the effects of PDE 3 inhibitors on preventing hair loss or promoting hair growth have not been reported yet.

Korean Patent No. 10-1820531

DISCLOSURE OF THE INVENTION The present inventors have conducted intensive studies on a method for promoting hair growth using a PDE 3 inhibitor including cilostazol which has been proven as a safety measure in order to solve the above conventional problems, (HDPC), which is a core cell for hair growth, and promotes hair growth in hair follicle organ culture and animal anagen induction, And the present invention has been completed on the basis thereof.

More particularly, the present invention relates to a pharmaceutical composition comprising as an active ingredient a PDE 3 inhibitor exhibiting an effect of preventing hair loss or accelerating hair growth, which composition comprises a PDE 3 inhibitor as an active ingredient, A cosmetic composition, and a quasi-drug composition.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing hair loss or promoting hair growth, which comprises a phosphodiesterase 3 inhibitor (PDE 3 inhibitor) as an active ingredient.

The PDE 3 inhibitor is preferably selected from the group consisting of Cilostazole, Amrinone (Inamrinone), Adibendan, Anagrelide, Benafentrine, Bucladesine, Carbazeran, Cilomilast, Cilostamide, Enoxamone, Fenspiride HCl, K-134 (1-cyclopropyl-1 - [(1R (2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), KMUP-1 (7- [2- [4- (2- chlorophenyl) piperazin-1-yl] ethyl] -1,3-dimethylpurine-2,6-dione, Luteolin, MKS492 (8-amino-3,7 dihydro-7- (2-methoxyethyl) , 3 dimethyl-1H-purine-2,6-dione), Meribendan, Milrinone, Olprinone, OPC-33540 (1-cyclooctyl- -2-hydroxycyclohexyl] -3- [3 - [(2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), ORG 9935 (3- (5,6-Dimethoxy- yl) -4-methyl-4,5-dihydro-1H-pyridazin-6-one, Parogrelil, Pimobend an), Pumafentrine, Quazinone, RPL-554 (2- [9,10-dimethoxy-4-oxo-2- (2,4,6-trimethylphenyl) imino- dihydropyrimido [6,1-a] isoquinolin-3-yl] ethylurea, Siguazodan, Salbutamol, Trequinsin, Vesnarinone, Zardaverine, etc. , But it is not limited thereto as long as it is an inhibitor capable of inhibiting the mechanism and / or activity of phosphodiesterase 3.

In one embodiment of the present invention, the composition may contain the PDE 3 inhibitor at a concentration of 0.0001 to 50 mM, but is not limited thereto.

In another embodiment of the present invention, the composition can be used for the prevention or treatment of alopecia. In particular, the alopecia is characterized by at least one of alopecia areata, hereditary alopecia, resting alopecia, traumatic alopecia, , Alopecia areata, alopecia alopecia, alopecia alopecia, alopecia areata, symptomatic alopecia, cicatric alopecia, and congenital alopecia.

In addition, the present invention provides a cosmetic composition for preventing hair loss or promoting hair growth comprising a PDE 3 inhibitor as an active ingredient.

In one embodiment of the present invention, the composition can be used for the prevention or improvement of alopecia. In particular, the alopecia can be used for alopecia areata, hereditary alopecia, resting alopecia, traumatic alopecia, But are not limited to, one or more members selected from the group consisting of non-rigid alopecia, alopecia alopecia, alopecia areata, symptomatic alopecia, cicatric alopecia, and congenital alopecia.

In another embodiment of the present invention, the composition may be formulated as an external preparation for skin such as a scalp treatment agent, a soap, a hair tonic, a shampoo, a rinse, a hair pack, a hair gel, a lotion, a conditioner, a hair oil, a mousse, But are not limited to, formulations of emulsions, emulsions, dispersants, micelles, liposomes, ointments, lotions, essences, patches, and sprays.

The present invention also provides a quasi-drug composition for preventing hair loss or promoting hair growth comprising a PDE 3 inhibitor as an active ingredient.

Furthermore, the present invention provides a method for preventing or treating alopecia, comprising treating or administering the above pharmaceutical composition to a subject.

In addition, the present invention provides the use of the pharmaceutical composition for the prevention or treatment of alopecia.

There are many kinds of hair-related products currently available on the market, but they are mostly ineffective in prevention of hair loss and hair growth, and they are temporary. Therefore, they do not meet the needs of users and there is also lack of evidence on its efficacy and safety Do. Furthermore, it has been reported that even in the case of minoxidil and propecia, which are effective in preventing hair loss, the alopecia can be recurred again when the product is discontinued, or serious side effects such as sexual dysfunction may be caused. . Accordingly, the inventors of the present invention have conducted intensive studies to develop drugs that have proved objective and reproducible effects on alopecia while having few side effects, and as a result, they have found that PDE 3 inhibitors exert excellent effects on hair loss or hair growth by promoting hair growth And completed the present invention. Cilostazol, which is a well-known PDE 3 inhibitor, has already been used as an antiplatelet agent and its pharmacological and pharmacokinetic properties have been clearly identified and proved to be safe. Therefore, according to the present invention, it can be used as a therapeutic agent for hair loss prevention, If you use it in the development of cosmetics, you will be able to see the savings in terms of cost rather than making a new drug at all, because it is a drug that is proven in terms of safety. .

1 is a graph showing the results of treatment of cilostazol, cilostamide, trequinsin and milrinone in human dermal papilla cells according to one embodiment of the present invention and performing BrdU assay PDE 3 < / RTI > inhibitor on the cell proliferation.
FIG. 2 is a graph showing the effect of a PDE 3 inhibitor according to an embodiment of the present invention on cell proliferation-related signal transduction pathways using Western blotting.
FIG. 3 is a graph showing the results of performing an ex vivo hair follicle organ culture test according to an embodiment of the present invention, wherein FIG. 3 a is a graph and a photograph showing follicular growth by cilostazol, 3b is a graph showing the result of performing immunofluorescence staining for Ki-67, a cell proliferation marker, in order to confirm cell proliferation in cultured hair follicles.
FIG. 4 is a graph showing the result of confirming the growth-inducing effect of cilostazol by the treatment of cilostazol and C57BL / 6 mouse model according to an embodiment of the present invention, to be.
FIG. 5 is a graph showing the results of analysis of growth shoot folliculogenesis by cilostazol according to an embodiment of the present invention. FIG. 5a is a graph showing the results of histological evaluation of growth factor hair follicle formation by hematoxylin and eosin staining. FIG. 5B is a graph showing anagen induction score by cilostazol, and FIG. 5C is a graph showing a change in skin thickness caused by cilostazol. FIG.
FIG. 6 is a graph showing the effect of cilostazol on cell proliferation-related signal transduction pathway using mouse skin tissue according to an embodiment of the present invention.
FIG. 7 shows the results of examining the effect of cilostazol according to an embodiment of the present invention on the peripheral vascular formation of hair follicles. FIG. 7A shows the result of staining of CD31, a blood vessel-labeled protein, ). Fig.

The inventors of the present invention have conducted intensive studies to develop drugs that have proven objective and reproducible effects on alopecia while having few side effects. As a result, it has been confirmed that PDE 3 inhibitors have an excellent effect on hair loss or hair growth by promoting hair growth, Thereby completing the present invention.

Hereinafter, the present invention will be described in detail.

The present invention provides a pharmaceutical composition for preventing hair loss or promoting hair growth, comprising a PDE 3 inhibitor as an active ingredient, wherein the composition can be used for the prevention or treatment of alopecia.

In one embodiment of the present invention, in order to evaluate the effect of phosphodiesterase (PDE) on proliferation of hair follicle cells, PDE 3 inhibitors cilostazol, cilostamide, Trequincin, and milrinone were treated, and BrdU and Western blotting were performed (see Examples 1 and 2).

In another embodiment of the present invention, in order to evaluate the effect of cilostazol on the growth of human hair follicles, an ex vivo hair follicle organ culture was performed, and the cells of the hair follicle cultured through the above- To confirm the proliferation, immunofluorescence staining was performed using a Ki-67 antibody as a cell division marker (see Example 3).

In another embodiment of the present invention, the effect of cilostazol on hair growth was confirmed using a C57BL / 6 mouse model, and the skin tissue growing on the growing hair was fixed with paraffin, and hematoxylin and eosin ), Histological examination was performed, and the degree of growth and skin thickness were measured (see Example 4).

In another embodiment of the present invention, in order to confirm the effect of cilostazol on the MAPK-related signal transduction pathway, which is a signal transduction pathway related to cell proliferation in mouse tissue, Western blotting is performed using mouse skin tissue (See Example 5).

In another embodiment of the present invention, immunofluorescence staining for CD31, an angioprotein, was performed to confirm the effect of cilostazol on angiogenesis around hair follicles (see Example 6).

In the present specification, the term "alopecia" refers to a state in which hair is not normally present at a site where hair should be present, and generally means that the hair of the scalp (coarse black hair) is missing. The Virgin Mary can cause cosmetic problems when she is out of color and has a different thickness. Korean people with lower hair density than Westerners have about 100,000 hairs and it is normal that about 50-100 hairs are lost per day. Hair loss can be divided into two types, that is, the formation of scars clinically and the formation of scars. The hair loss in which the scar is formed does not regenerate the hair because the hair follicle is destroyed, while the hair loss in which the scar is not formed maintains the hair follicle The hair is regenerated after the site has disappeared. Non-scarring hair loss without scarring includes hereditary androgenetic alopecia (baldness), alopecia areata, hair folliculitis caused by fungal infection, hair resting hair, hairy wall, hair growth disorder disease and scarring hair loss disorder Hair loss caused by lupus, poisonous folliculitis, papillary flattening, hair loss due to burns and trauma. Among the alopecia, the most frequent is alopecia herpeticus and alopecia areata, and they all have no scarring. Among the diseases with high frequency of hair loss diseases are baldness (male hair loss), female hair loss, alopecia areata, resting hair alopecia. In addition, the alopecia of the present invention may include all the problems due to the phenomenon that a large number of hairs are missing, such as cosmetic problems, in addition to the above-mentioned diseases.

As used herein, the term "phosphodiesterase (PDE)" refers to phosphodiesterase (PDE) derived from phosphoric acid diesters such as cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate Is a kind of phosphatase that decomposes the bond. It is an enzyme that catalyzes the reaction of forming monoester and alcohol through hydrolysis. It is classified into 12 subtypes. The PDE regulates the size, location and persistence of cyclic nucleotide signaling within the sub-domain of the cell, and plays an important role in the regulation of signal transduction mediated by various secondary messengers . Because of the unique tissue distribution and structural and functional characteristics of such PDE enzymes, inhibition of PDE inhibits hydrolysis of PDE, thereby affecting various physiological processes mediated by cAMP, cGMP, etc., Is used as a suppression target, and is actively studied.

As used herein, the term "phosphodiesterase 3 inhibitor (PDE 3 inhibitor)" refers to a substance that selectively inhibits the enzyme of subtype 3 of the phosphodiesterase enzyme, and is used for improving myocardial contractility and vasodilation Has been studied extensively as a cardiovascular drug. Among these inhibitors, " cilostazol " is a quinolinone derivative, which is one of the PDE 3 inhibitors widely used in the clinical field, and increases intracellular cAMP levels in vascular smooth muscle cells , And is used safely for the elderly as an antiplatelet agent. Its pharmacological and pharmacokinetic properties are clearly defined, and it is currently marketed for the purpose of improving ischemic symptoms such as ulceration, pain and cold sensation caused by chronic arterial occlusion and inhibiting recurrence after cerebral infarction It is a widely used drug. In addition to cilostazol, PDE 3 inhibitors include amrinone (Inamrinone), Adibendan, Anagrelide, Benafentrine, Bucladesine, Carbazeran, Cilomilast, Cilostamide, Enoxamone, Fenspiride HCl, K-134 (1-cyclopropyl-1 - [(1R, 2R) (7- [2- [4- (2-chlorophenyl) piperazin-1-yl] -hydroxycyclohexyl] -3- [3- -yl] ethyl] -1,3-dimethylpurine-2,6-dione, Luteolin, MKS492 (8-amino-3,7 dihydro-7- (2-methoxyethyl) 2-hydroxycyclohexyl] -purine-2,6-dione, Meribendan, Milrinone, Olprinone, OPC-33540 (1-cyclooctyl- 3- [3 - [(2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), ORG 9935 (3- (5,6-Dimethoxy- 1- benzothiophen- methyl-4,5-dihydro-1H-pyridazin-6-one, Parogrelil, Pimobendan, 4-oxo-2- (2,4,6-trimethylphenyl) imino-6,7-dihydropyrimido [6,1- a isoquinolin-3-yl] ethylurea, Siguazodan, Salbutamol, Trequinsin, Vesnarinone, Zardaverine and the like. The present invention is not limited thereto as long as it is a substance that can specifically inhibit the activity of PDE 3.

As used herein, the term " prevention " means any action that inhibits or delays disease caused by administration of the composition according to the present invention. Inhibiting or delaying the onset of hair loss may be considered as hair loss, that is, hair loss is suppressed, hair growth (hair is generated or grown) is promoted, or hair loss suppression and hair growth promotion are simultaneously performed, Means action.

In the present specification, the term " treatment " means any action that alleviates or alleviates symptoms such as hair loss-related diseases by administration of the composition according to the present invention.

As used herein, " improvement " means an act of at least reducing the degree of symptom associated with the condition being treated, for example. At this time, the composition of the present invention can be used simultaneously or separately as a medicament for treatment for alopecia prevention or improvement.

As used herein, the term " individual " refers to a subject to which the composition of the present invention can be administered, and there is no limitation to the subject.

As used herein, the term "pharmaceutical composition" means a capsule, a tablet, a granule, an injection, an ointment, a powder or a beverage, and the pharmaceutical composition is intended for a human being . The pharmaceutical composition may be formulated in the form of oral preparations such as powders, granules, capsules, tablets, aqueous suspensions, external preparations, suppositories, and sterilized injection solutions according to conventional methods, though not limited thereto . The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier may be a binder, a lubricant, a disintegrant, an excipient, a solubilizing agent, a dispersing agent, a stabilizer, a suspending agent, a coloring matter, a perfume or the like in the case of oral administration. A solubilizing agent, an isotonic agent, a stabilizer and the like may be mixed and used. In the case of topical administration, a base, an excipient, a lubricant, a preservative and the like may be used. Formulations of the pharmaceutical compositions of the present invention may be prepared in a variety of ways by mixing with pharmaceutically acceptable carriers as described above. For example, oral administration may be in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc. In the case of injections, they may be formulated in unit dosage ampoules or in multiple dosage forms have. Other, solutions, suspensions, tablets, capsules, sustained-release preparations and the like.

Examples of suitable carriers, excipients and diluents for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltoditol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate or mineral oil. Further, it may further include a filler, an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent, an antiseptic, and the like.

The route of administration of the pharmaceutical composition according to the present invention may be, but is not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, , Sublingual or rectal. Oral or parenteral administration is preferred. The term "parenteral" as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional and intracranial injection or infusion techniques. The pharmaceutical compositions of the present invention may also be administered in the form of suppositories for rectal administration.

The pharmaceutical composition of the present invention may be administered orally or parenterally depending on various factors including the activity of the specific compound used, age, weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination, And the dosage of the pharmaceutical composition may be appropriately selected by a person skilled in the art depending on the condition of the patient, the body weight, the degree of disease, the type of drug, the route of administration and the period of time, and is preferably from 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way. The pharmaceutical composition according to the present invention can be formulated into pills, dragees, capsules, solutions, gels, syrups, slurries, and suspensions.

In the present invention, the cosmetic composition may contain 0.0005 to 50% by weight, based on the total weight of the composition, of the inhibitor of the final glycocalyx receptor. In addition, the composition of the present invention may contain at least one active ingredient that exhibits the same or similar function in addition to the inhibitor of the final glycocalyx receptor.

As used herein, the term " cosmetic composition " may be used in the form of a general emulsified formulation and a solubilized formulation, and the cosmetic composition may be a human. Cosmetics manufactured by the cosmetic composition of the present invention can be produced in the form of a general emulsified formulation and a solubilized formulation. Examples of cosmetics of the emulsified formulation include nutritional lotion, cream, essence and the like, and the cosmetics of the solubilized formulation have flexibility . It can also be prepared in the form of adjuvants which can be applied topically or systemically, which are conventionally used in the field of dermatology by containing a dermatologically acceptable medium or base.

Suitable cosmetic formulations include, for example, solutions, gels, solid or kneaded anhydrous products, emulsions obtained by dispersing the oil phase in water, suspensions, microemulsions, microcapsules, microgranules or ionic (liposomes) In the form of a foliar dispersant, in the form of a cream, a skin, a lotion, a powder, an ointment, a spray or a conceal stick. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

The cosmetic composition of the present invention may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softening agents, antioxidants, suspending agents, stabilizers, foaming agents, perfumes, Or any other conventionally used in cosmetics such as nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, ≪ RTI ID = 0.0 > cosmetics < / RTI > such as botanical ingredients, or adjuvants conventionally used in the field of dermatology. And the above ingredients may be introduced in amounts commonly used in the dermatology field.

Examples of products to which the cosmetic composition of the present invention can be added include cosmetics such as convergent lotion, softening longevity, nutritional lotion, various creams, essences, packs and foundations, cleansing, cleanser, soap, , Essence, essence and so on.

Specific formulations of the cosmetic composition of the present invention include hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, Hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair drying agent, hair preservative, hair dye, hair wave agent, hair decolorizer, hair gel, hair glaze, hair dresser, hair lacquer , Hair Moisturizer, Hair Moisture Hair Spray, Skin Lotion, Skin Softener, Skin Toner, Astringent, Lotion, Milk Lotion, Moisture Lotion, Nutrition Lotion, Massage Cream, Nourishing Cream, Moisture Cream, Hand Cream, Essence, Nutrition Essence Pack , Soap, Shampoo, Cleansing Foam, Cleansing Lotion, Cleansing Cream, Body Lotion, Body Cleanser, Milk, Press Powder, Loose Powder, Eye Shadow The formulation comprises a.

When the formulation of the present invention is a paste, cream or gel, an animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as the carrier component .

When the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. In the case of a spray, in particular, / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters.

The ingredients contained in the cosmetic composition of the present invention include, in addition to the active ingredient and the carrier ingredient, ingredients conventionally used in cosmetic compositions and include conventional additives such as antioxidants, stabilizers, solubilizers, vitamins, . ≪ / RTI >

In the present specification, the term "quasi-drug" refers to a product used for the purpose of treating or alleviating, treating, or preventing a disease or a disease of a human being or an animal, wherein the action on the human body is mild or does not act directly on the human body.

Hereinafter, the following examples are provided to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

Example

Example  One. PDE  Derived from human hair follicles by 3 inhibitors Dermal papilla  Induction of cell division

To evaluate the effect of phosphodiesterase (PDE) on the proliferation of hair follicle cells, the well known PDE 3 inhibitors cilostazol, cilostamide, trequincin, And BrdU analysis was performed.

First, the primary cultured dermal papilla cells were treated with cilostazol, cilostamide, trequincin, and milrinone at a concentration of 0 μM to 10 μM, cultured for 44 hours, treated with BrdU reagent to the dermal papilla cells, Lt; / RTI > After the final 48 hours of incubation, the luminescence was measured using a luminometer for detecting BrdU, and the degree of cell division and activation was confirmed by comparing with the negative control. The results are shown in Fig.

As shown in Fig. 1, it was confirmed that cilostazol, cilostamide, trequincin, and milrinone significantly increased the cell division of the dermal papilla cells (hDPC). From the above results, it was confirmed that the PDE 3 inhibitors significantly increase the cell division of the dermal papilla cells by inhibiting the activity of PDE 3, and thus, the hair growth promoting effect can be demonstrated.

Example  2: derived from human hair follicle In dermal papilla cells PDE  3 inhibitor ERK  Identification of signal transduction protein phosphorylation regulation

In order to examine the effect of PDE 3 inhibitors on extracellular-signal-regulated kinase (ERK) signaling pathway, a key signaling pathway involved in cell proliferation, cilostazol, cilostamide, trequincin, Were treated with 1 μM each for 0, 5, 15, and 60 minutes, respectively, and Western blotting was performed. The results are shown in Fig.

As shown in FIG. 2, it was confirmed that the phosphorylation of intracellular ERK protein was increased within 15 minutes by cilostazol, cilostamide, trequincin, and milrinone, and it was confirmed that PDE 3 inhibitors inhibited the ERK signaling pathway . ≪ / RTI >

Example  3: Through human follicular organ culture test PDE  3 Inhibitors Promote Human Hair Loss Growth

In order to evaluate the effect of PDE 3 inhibitor on human hair growth, ex vivo hair follicle organ culture was performed using cilostazol. More specifically, in order to perform a human follicular organ culture test, the scalp tissue obtained from volunteers was separated into follicular units, and the hair follicles remaining in the lower part of the sebaceous gland of the isolated follicles were used. The length of the hair growing on the 2nd, 4th, and 6th days after the addition of the culture medium containing the negative control reagent or cilostazol 1 and 10 μM to the prepared hair follicles was measured and compared with the negative control, Of the hair growth promoting effect. The results are shown in Fig.

As shown in Fig. 3A, it was confirmed that hair growth of human hair follicle was promoted by cilostazol.

In order to confirm the cell proliferation of the hair follicles cultured through the above test, immunofluorescence staining was performed using a Ki-67 antibody as a cell division marker. The hair follicles treated with cilostazol and incubated for two days were cut into 4 μm thick slices and fixed on a slide. Ki-67 was treated with the primary antibody, and green fluorescence capable of detecting Ki-67 was bound After the secondary antibody treatment, the cells were observed with a fluorescence microscope and photographed to confirm the number of Ki-67 positive cells. Nuclei were stained with DAPI reagent. The result is shown in FIG. 3B.

As shown in FIG. 3B, it was confirmed that cell division was increased in human hair follicle by cilostazol.

Example 4: Growth of hair by PDE 3 inhibitor

In order to confirm the hair growth effect by the PDE 3 inhibitor, the C57BL / 6 mouse model was used to confirm the growth-inducing effect. Although human hairs have different hair cycles depending on individuals, all of the hairs are initially in the same cycle, and when they are 7 to 8 weeks of age, all of the hairs enter the resting period and thereafter are converted into the growth period. In order to evaluate the effect of inducing the growth of the plants, the hair was removed from the C57BL / 6 mouse at 8 weeks of age, which was in a resting state, and then 0.2wt% and 1wt% of cilostazol were applied once a day for 3 weeks. The hair growth and the induction of the growth of the hair were confirmed by comparing the control (Control) with 2% by weight (Minoxidil 2% by weight) of the positive control group, minoxidil. The results are shown in Fig.

As shown in FIG. 4, it was confirmed that the ratio of the growth area of the growth hair was significantly increased in the cilostazol-treated group, and it was confirmed that the PDE 3 inhibitor promoted the growth of hair.

In addition, the results of histological examination by fixing the skin tissue of the growing hair with paraffin and staining with hematoxylin and eosin are shown in FIG.

As shown in Fig. 5A, it was confirmed that hair follicle formation was promoted in the cilostazol-treated experimental group. As shown in Figs. 5B and 5C, in the experimental group treated with cilostazol, the Anagen induction score and Skin thickness was also increased. In addition, in the case of cilostazol, it was confirmed that even at a concentration of 0.2% lower than that of minoxidil (MNX2%), the effect was similar or increased to that of minoxidil.

Example  5: PDE  3 inhibitor MAPK  Identification of phosphorylation regulation of related signaling proteins

Western blot was performed using mouse skin tissue to confirm the effect of PDE 3 inhibitor on MAPK-related signal transduction pathway involved in cell proliferation. The protein was extracted from mouse skin tissue subjected to the growth induction experiment by applying negative control (Control; Con), cilostazol (CIL), and positive control group, minoxidil (MNX) The MAPK-related proteins extracellular signal-regulated kinases (ERK), c-Jun N-terminal kinases (JNK) and phosphorylation of P-38 were confirmed by Western blotting. The results are shown in Fig.

As shown in Fig. 6, it was confirmed that phosphorylation of MAPK-related signal transduction protein was increased by cilostazol.

Example  6: PDE  3 inhibitor induces angiogenesis induction

In order to confirm the effect of PDE 3 inhibitor on the angiogenesis around hair follicles, immunofluorescence staining for CD31, an angioprotein, was performed. More specifically, the mouse skin tissues coated with the negative control and cilostazol were rapidly frozen to prepare blocks, which were cut into 15 mu m thick tissue slices using a cryotome and fixed on the slides. Then, the primary antibody is treated with CD31 antibody, which is an angioprotein, and then the secondary antibody with green fluorescence capable of detecting CD31 is treated, and the image is observed with a fluorescence microscope. The area was measured. The results are shown in Fig.

As shown in FIG. 7A, angiogenesis was induced by cilostazol, and the area of blood vessels was also increased as shown in FIG. 7B.

Through the above results, the PDE 3 inhibitors of the present invention inhibit PDE 3 activity in the scalp and activate phosphorylation of ERK protein and MAPK-related signal transduction proteins, thereby activating the proteins, thereby promoting cleavage and vascularization of the papillary cells Finally, it was confirmed that the growth of hair was promoted and the growth of hair was promoted to exhibit the effect of hair growth. Through this, it was confirmed that hair loss can be prevented or suppressed, or foaming can be promoted, .

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention. There will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

Claims (12)

A pharmaceutical composition for preventing hair loss or promoting hair growth, comprising as an active ingredient a phosphodiesterase 3 inhibitor (PDE 3 inhibitor). The method according to claim 1,
The phosphodiesterase 3 inhibitor may be selected from the group consisting of Cilostazole, Amrinone (Inamrinone), Adibendan, Anagrelide, Benafentrine, Bucladesine ), Carbazeran, Cilomilast, Cilostamide, Enoxamone, Fenspiride HCl, K-134 (1-cyclopropyl-1 - [ (7- [2- [4- (2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), KMUP- -chlorophenyl) piperazin-1-yl] ethyl] -1,3-dimethylpurine-2,6-dione, Luteolin, MKS492 (8-amino-3,7 dihydro- 1,3 dimethyl-1H-purine-2,6-dione, Meribendan, Milrinone, Olprinone, OPC-33540 (1-cyclooctyl- ) -2-hydroxycyclohexyl] -3- [3 - [(2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), ORG 9935 (3- (5,6-dimethoxy- -yl) -4-methyl-4,5-dihydro-1H-pyridazin-6-one, parogrelil, imobendan, pumafentrine, quazinone, 2- [9,10-dimethoxy-4-oxo-2- (2,4,6-trimethylphenyl) imino- dihydropyrimido [6,1-a] isoquinolin-3-yl] ethylurea, Siguazodan, Salbutamol, Trequinsin, Vesnarinone, and Zardaverine. ≪ / RTI > or a pharmaceutically acceptable salt thereof. The method according to claim 1,
Wherein the composition is characterized in that the phosphodiesterase 3 inhibitor is contained in a concentration of 0.0001 to 50 mM. The method according to claim 1,
Wherein said composition is for the prevention or treatment of alopecia. 5. The method of claim 4,
The alopecia is caused by alopecia areata, hereditary androgenetic alopecia, resting alopecia areata, traumatic alopecia, hairy wall, compression alopecia, alopecia alopecia, non-rigorous alopecia, alopecia alopecia, alopecia alba, symptomatic alopecia and congenital alopecia Lt; RTI ID = 0.0 > 1, < / RTI > A cosmetic composition for preventing hair loss or promoting hair growth, comprising as an active ingredient a phosphodiesterase 3 inhibitor (PDE 3 inhibitor). The method according to claim 6,
The phosphodiesterase 3 inhibitor may be selected from the group consisting of Cilostazole, Amrinone (Inamrinone), Adibendan, Anagrelide, Benafentrine, Bucladesine ), Carbazeran, Cilomilast, Cilostamide, Enoxamone, Fenspiride HCl, K-134 (1-cyclopropyl-1 - [ (7- [2- [4- (2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), KMUP- -chlorophenyl) piperazin-1-yl] ethyl] -1,3-dimethylpurine-2,6-dione, Luteolin, MKS492 (8-amino-3,7 dihydro- 1,3 dimethyl-1H-purine-2,6-dione, Meribendan, Milrinone, Olprinone, OPC-33540 (1-cyclooctyl- ) -2-hydroxycyclohexyl] -3- [3 - [(2-oxo-1H-quinolin-6-yl) oxy] propyl] urea), ORG 9935 (3- (5,6-dimethoxy- -yl) -4-methyl-4,5-dihydro-1H-pyridazin-6-one, parogrelil, imobendan, pumafentrine, quazinone, 2- [9,10-dimethoxy-4-oxo-2- (2,4,6-trimethylphenyl) imino- dihydropyrimido [6,1-a] isoquinolin-3-yl] ethylurea, Siguazodan, Salbutamol, Trequinsin, Vesnarinone, and Zardaverine. Wherein the cosmetic composition is at least one selected from the group consisting of: The method according to claim 6,
Wherein the composition contains phosphodiesterase 3 inhibitor at a concentration of 0.0001 to 50 mM. The method according to claim 6,
Wherein the composition is for preventing or ameliorating alopecia. 10. The method of claim 9,
The alopecia is caused by alopecia areata, hereditary androgenetic alopecia, resting alopecia areata, traumatic alopecia, hairy wall, compression alopecia, alopecia alopecia, non-rigorous alopecia, alopecia alopecia, alopecia alba, symptomatic alopecia and congenital alopecia Or a pharmaceutically acceptable salt thereof. The method according to claim 6,
The composition may be formulated as an external preparation for skin such as a scalp treatment agent, a soap, a hair tonic, a shampoo, a rinse, a hair pack, a hair gel, a lotion, a conditioner, a hair oil, a mousse, a cream, , An ointment agent, a lotion, an essence, a patch, and a spray agent. A quasi-drug composition for preventing hair loss or promoting hair growth, comprising as an active ingredient a phosphodiesterase 3 inhibitor (PDE 3 inhibitor).

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