WO2023191515A1 - Composition for preventing hair loss or promoting hair growth comprising indirubin derivative and metabolic activator - Google Patents

Abstract

The present invention relates to a composition for preventing hair loss or promoting hair growth, comprising an indirubin derivative and a metabolic activator. The composition, according to the present invention, may be effectively used as a pharmaceutical composition or a cosmetic composition that may prevent hair loss or promote hair growth, and may be used in functional cosmetics with hair growth promotion or hair loss prevention properties exhibited from a hair loss prevention function, including a hair growth promoter.

Description

Composition for preventing hair loss or promoting hair growth containing an indirubin derivative and a metabolic activator

The present invention relates to a composition for preventing hair loss or promoting hair growth containing an indirubin derivative and a metabolic activator.

There are about 100,000 to 150,000 hairs in the human body, and they are formed in hair follicles. Each hair cycle repeats the growth phase (anagen), the catagen phase, and the resting phase (telogen) in different cycles. The anagen phase is the period when hair grows, and the catagen phase is the period when the metabolic process slows down while maintaining the shape of the hair after the growth period ends. During this period, hair growth slows down. The catagen stage is a period in which hair follicle cell division stops and rapidly shrinks. The final telogen phase is a period in which hair follicle activity completely stops and prepares for hair loss. The telogen hair is pushed out by new growth-phase hair growing at the base and falls out on its own, and is easily removed by physical actions such as combing or washing the hair. This cycle repeats over 3 to 6 years.

Hair loss refers to a condition in which there is no hair in areas where hair should normally exist. As the hair loss disease progresses, the papilla present at the hair root becomes smaller, and as the papilla becomes smaller, the thickness of the hair becomes thinner and at the same time, the hair cycle becomes shorter and new growth occurs. The hair that comes out becomes thinner. In other words, as hair loss progresses, the hair turns into fluff and the hair cycle becomes shorter, so the hair grows a little and falls out immediately.

It is known that the causes of hair loss are not only genetic causes and the action of male hormones, but also a combination of factors such as endocrine diseases, nutritional deficiencies, drug use, childbirth, fever, and severe physical and mental stress such as surgery. Among them, male pattern hair loss is known to be greatly influenced by dihydrotestosterone (DHT). Testosterone, a type of male hormone, is activated into DHT by the 5α-reductase enzyme, and this DHT is known to induce hair loss. DHT causes hair follicles to atrophy, causing vellus hair to become thin and limp. Alternatively, hair root-destroying substances are secreted, causing the hair to enter the catagen stage at an early stage, resulting in hair loss.

Hair loss treatments developed to date include finasteride (brand name: Propecia®), dutasteride (brand name: Avodart®), and minoxidil (brand names: Minoxil® or Rogaine®). Minoxidil can cause side effects such as edema, arrhythmia, and hair growth in unwanted areas when applied long-term. It is known that the effect of minoxidil is greatest between 6 months and 1 year after use, and the effect gradually decreases thereafter. . Finasteride and dutasteride inhibit DHT production by blocking the action of the 5α-reductase enzyme, but have side effects such as decreased libido, erectile dysfunction, and loss of driving and performance abilities. In addition, hair transplant surgery has recently been attempted for patients with severe hair loss, but the high cost and side effects after the procedure have been pointed out as limitations.

Against this background, the present inventors conducted repeated research to develop a drug with fewer side effects and excellent hair loss improvement effect, and as a result discovered that indirubin derivatives and metabolic activators have excellent hair loss improvement effect, thereby completing the present invention. .

Patent Document 1. Republic of Korea Patent Publication No. 10-2022-0032974

The present invention was devised to solve the above problems, and the object of the present invention is to provide a pharmaceutical composition for preventing or treating hair loss that contains an indirubin derivative and a metabolic activator as active ingredients and has no side effects on the human body. will be. In addition, the present invention provides a cosmetic composition and food composition for preventing or improving hair loss containing the indirubin derivative and the metabolic activator as active ingredients.

Another object of the present invention is to provide a method for preventing, improving, or treating hair loss that has an excellent hair loss prevention or treatment effect and includes an indirubin derivative and a metabolic activator as active ingredients.

In addition, the present invention is intended to provide a new use of a composition containing an indirubin derivative and a metabolic activator for the prevention or treatment of hair loss.

In order to achieve the above object, the present invention includes an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients, and the metabolic activator is a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate carrier) Provided is a pharmaceutical composition for preventing or treating hair loss, which is an inhibitor.

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

In Formula 1, R ₁ to R ₃ are each independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₆ alkyl, and C ₁ -C ₆ alkoxy, and R ₄ is hydrogen or C ₁ - C ₆ alkyl, and R ₅ may be any one selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 arylalkyl.

In Formula 1, R ₃ and R ₄ may be hydrogen.

In Formula 1, R ₁ and R ₂ may each independently be selected from the group consisting of hydrogen, halogen, and -O-CH ₃ .

In Formula 1, R ₅ may be any one selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-10 arylalkyl.

Formula 1 may be any one selected from the following Formulas 1-1 to 1-4.

[Formula 1-1]

[Formula 1-2]

[Formula 1-3]

[Formula 1-4]

The PKM2 (pyruvate kinase M2) activator is 6-[(3-aminophenyl)methyl]-4,6-dihydro-4-methyl-2-(methylsulfinyl)-5H-thieno[2',3':4,5 ]pyrrolo[2,3-d]pyridazin-5-one(TEPP-46, ML265), 3-(4-(2,3-Dihydrobenzo[b][1,4]dioxin-6-ylsulfonyl)-1, It may be any one selected from the group consisting of 4-diazepan-1-ylsulfonyl)aniline (DASA-58) and serine, and the MPC (Mitochondrial pyruvate carrier) inhibitor is (E)-2-Cyano-3- It may be (1-phenyl-1H-indol-3-yl)acrylic acid (UK5099).

The indirubin derivative and the metabolic activator may be included in a molar ratio of 1:0.1 to 10.

In order to achieve the above other objects, the present invention includes an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients, and the metabolic activator is a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate carrier) ) Provides a cosmetic composition for preventing or improving hair loss, characterized in that it is an inhibitor.

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

The composition may promote the proliferation of dermal papilla cells or hair follicle cells in vitro.

The cosmetic composition includes 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, and hair aerosol. , hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair dryer, hair preservation treatment, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, It may be any one hair formulation selected from the group consisting of hair mousse and hair spray.

In order to achieve the above other object, the present invention includes an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients, and the metabolic activator is a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate) Provided is a food composition for preventing or improving hair loss, which is characterized as being a carrier inhibitor.

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

In order to achieve the above further object, the present invention provides a method for preventing or treating hair loss, which includes administering an indirubin derivative represented by the following formula (1) and a metabolic activator to a hair loss patient.

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

In order to achieve the above-mentioned other object, the present invention provides a new use of an indirubin derivative and a metabolic activator represented by the following formula (1) as a drug for preventing or treating hair loss. The metabolic activator is characterized as being a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate carrier) inhibitor.

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

In order to achieve the above-mentioned further object, the present invention provides a method for producing a pharmaceutical agent having a hair loss prevention or treatment effect, characterized by using an indirubin derivative and a metabolic activator represented by the following formula (1): .

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

The composition according to the present invention can be usefully used as a pharmaceutical composition or cosmetic composition capable of preventing hair loss or promoting hair growth, and can be used in functional cosmetics for preventing hair loss or promoting hair growth, including hair growth promoting agents.

In addition, the mixed composition of an indirubin derivative and a metabolic activator according to the present invention has the advantage of being a safe material that is non-toxic to cells and does not cause inflammation and does not cause side effects to the human body.

The present invention is a composition that mixes an indirubin derivative and a metabolic activator at a certain ratio, and has the advantage of showing a stronger effect on hair growth, wool growth, and hair loss prevention than a single composition.

Figure 1 is a micrograph taken after culturing Vibrissa hair follicles treated with indirubin derivatives or metabolic activators (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) alone for 6 days, and Figure 2 is This is a photomicrograph taken after culturing Vibrissa hair follicles treated with a mixture of indirubin derivatives and metabolic activators (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) for 6 days.

Figure 3 shows the length change of each Vibrissa hair follicle treated alone or in combination with an indirubin derivative and a metabolic activator (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) and cultured for 6 days. This is the graph shown (* P < 0.05, ** P < 0.005, *** P < 0.0005, NS: not significant vs. VEH or single treat).

Figure 4 is a micrograph (top) of Vibrissa hair follicles taken in the group administered with various concentrations of serine alone and a graph (bottom) showing the length change (* P < 0.05, ** P < 0.005, *** P < 0.0005, NS: not significant vs. VEH).

Figure 5 shows the cell viability of human dermal papilla cells treated with indirubin derivatives and metabolic activators (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) alone or in combination. This is the graph shown (* P < 0.05, ** P < 0.005, *** P < 0.0005 vs VEH).

Figure 6 is a photograph of each group treated with indirubin derivative and metabolic activator (TEPP-46) alone or in combination, and Figure 7 is a photograph of each group treated with indirubin derivative and metabolic activator (TEPP-46) alone or in combination. This is a graph showing the hair growth values measured from each treated group (* P < 0.05, ** P < 0.005, *** P < 0.0005, NS: not significant vs VEH or single treat).

Below, we will look at various aspects and various implementation examples of the present invention in more detail.

Throughout the specification of the present application, when a part "includes" a certain component, this means that it may further include other components rather than excluding other components unless specifically stated to the contrary.

One aspect of the present invention relates to a pharmaceutical composition, cosmetic composition, and food composition for preventing or treating hair loss containing an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients. The metabolic activator may be a PKM2 (pyruvate kinase M2) activator or an MPC (Mitochondrial pyruvate carrier) inhibitor.

Another aspect of the present invention relates to a method of treating hair loss comprising administering an effective amount of an indirubin derivative represented by Formula 1 and a metabolic activator to a hair loss patient.

Another aspect of the present invention relates to a method of treating hair loss comprising administering a therapeutically effective amount of an indirubin derivative represented by Formula 1 and a metabolic activator to a subject.

In addition, another aspect of the present invention provides a new use of the indirubin derivative and metabolic activator represented by the following formula (1) as a drug for preventing or treating hair loss.

In addition, another aspect of the present invention provides a method for producing a pharmaceutical agent having a hair loss prevention or treatment effect, characterized by using an indirubin derivative represented by the following formula (1) and a metabolic activator.

In the present invention, the indirubin derivative and metabolic activator represented by Formula 1 are active ingredients that enable the above method and use to be achieved, and may be in the form of a mixture or mixed composition or composition mixed in a certain ratio.

The present invention is a composition that mixes an indirubin derivative and a metabolic activator in a certain ratio, and has been confirmed to have excellent effects on preventing hair loss and promoting hair growth, making it possible to prevent hair loss and promote hair growth, and thus can be used in pharmaceuticals, foods, cosmetics, etc. It can be applied to various fields.

[Formula 1]

In Formula 1,

R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

The term 'halogen' in the present invention refers to bromo, chloro, fluoro, or iodo, and can be used interchangeably with 'halo' and 'halogen'.

The term 'alkyl' in the present invention refers to a hydrocarbon having primary, secondary, tertiary, and/or quaternary carbon atoms, and includes saturated aliphatic groups that may be straight-chain, branched, or cyclic, or a combination thereof. do. For example, an alkyl group may have 1 to 20 carbon atoms (i.e., C ₁ -C ₂₀ alkyl), 1 to 10 carbon atoms (i.e., C ₁ -C ₁₀ alkyl), or 1 to 6 carbon atoms (i.e., C ₁ -C ₆ alkyl). Examples of suitable alkyls include methyl (Me, -CH ₃ ), ethyl (Et, -C ₂ H ₅ ), 1-propyl (n-Pr, -CH ₂ CH ₂ CH ₃ ), 2-propyl (i-Pr, -CH(CH ₃ ) ₂ ), 1-butyl (n-Bu, - CH ₂ CH ₂ CH ₂ CH ₃ ), 2-methyl-1-propyl (i-Bu,-CH ₂ CH(CH ₃ ) ₂ ) , 2-butyl (s-Bu, -CH(CH ₃ )CH ₂ CH ₃ ), 2-methyl-2-propyl (t-Bu, -C(CH ₃ ) ₃ ), 1-pentyl (n-pentyl, -CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ), 2-pentyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₃ ), 3-pentyl (-CH(CH ₂ CH ₃ ) ₂ ), 2-methyl- 2-Butyl(-CH(CH ₃ ) ₂ CH ₂ CH ₃ ), 3-methyl-2-butyl(-CH(CH ₃ )CH(CH ₃ ) ₂ ), 3-methyl-1-butyl(-CH ₂ CH ₂ CH(CH ₃ ) ₂ ), 2-methyl-1-butyl (-CH ₂ CH(CH ₃ )CH ₂ CH ₃ ), 1-hexyl (-CH ₂ CH ₂ CH ₂ CH ₂ CH ₂ CH ₃ ) , 2-hexyl (-CH(CH ₃ )CH ₂ CH ₂ CH ₂ CH ₃ ), 3-hexyl (-CH(CH ₂ CH ₃ )(CH ₂ CH ₂ CH ₃ )), 2-methyl-2-pentyl (-C(CH ₃ ) ₂ CH ₂ CH ₂ CH ₃ ), 3-methyl-2-pentyl(-CH(CH ₃ )CH(CH ₃ )CH ₂ CH ₃ ), 4-methyl-2-pentyl(- CH(CH ₃ )CH ₂ CH(CH ₃ ) ₂ ), 3-methyl-3-pentyl(-C(CH ₃ )(CH ₂ CH ₃ ) ₂ ), 2-methyl-3-pentyl(-CH(CH ₂ CH ₃ )CH(CH ₃ ) ₂ ), 2,3-dimethyl-2-butyl (-C(CH ₃ ) ₂ CH(CH ₃ ) ₂ ), 3,3-dimethyl2-butyl (-CH(CH ₃ )C(CH ₃ ) ₃ ), and octyl (-(CH ₂ ) ₇ CH ₃ ), preferably a C ₁ -C ₆ alkyl group, more preferably C ₁ -C ₃ The alkyl group may be any one selected from methyl (Me, -CH ₃ ), ethyl (Et, -C ₂ H ₅ ), and 1-propyl (n-Pr, -CH ₂ CH ₂ CH ₃ ).

As described above, the term 'alkoxy' in the present invention refers to a functional group with -OR in which an alkyl group is attached to the parent compound through an oxygen atom. Examples of C ₁ -C ₆ alkoxy include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, pentoxy, and hexoxy.

In the present invention, the term 'aryl', unless otherwise specified, refers to an optionally substituted benzene ring, or a ring system that can be formed by fusing one or more optional substituents, and the number of carbon atoms is not particularly limited, but is 6 to 6. Substituted or unsubstituted allyl, which is 30, is preferred. Examples of 'aryl' groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, biphenyl, indanyl, anthracyl or phenanthryl, and substituted derivatives thereof.

The substituents include C _l - ₃ alkyl, C ₂ - ₃ alkenyl, substituted C ₂ - ₃ alkynyl, heteroaryl, heterocyclic, aryl, alkoxy, aryloxy, aralkoxy, optionally having 1 to 3 fluorine substituents. , acyl, aroyl, heteroaroyl, acyloxy, aroyloxy, heteroaroyloxy, sulfanyl, sulfinyl, sulfonyl, aminosulfonyl, sulfonylamino, carboxyamide, aminocarbonyl, carboxy, oxo, Includes hydroxy, mercapto, amino, nitro, halogen or ureido. This ring or ring system may optionally be fused to an aryl ring (eg, a benzene ring), a carbocyclic ring, or a heterocyclic ring, optionally bearing one or more substituents. More preferably, the substituent may be C _1-3 alkyl, and aryl substituted with C _1-3 alkyl refers to an aryl group in which one hydrogen is substituted with an alkyl group. The aryl group substituted with C _1-3 alkyl is also called alkylaryl and may be C ₇ to C ₃₀ alkylaryl. Specifically, C _7-30 alkylaryl may be methylphenyl, ethylphenyl, n-propylphenyl, iso-propylphenyl, n-butylphenyl, iso-butylphenyl, tert-butylphenyl, or cyclohexylphenyl, and is preferably C It may be ₇ to C ₁₀ alkylaryl.

In the present invention, the arylalkyl group is arylalkyl having 7 to 30 carbon atoms, which may mean a substituent in which one or more hydrogens of alkyl are replaced by aryl. Specifically, it may be C _7-30 arylalkyl, preferably benzyl, phenylpropyl, or phenylhexyl, and more preferably, the C _7-10 arylalkyl may be benzyl or phenylpropyl.

The term 'hydroxy' in the present invention refers to the -OH radical.

In Formula 1, R ₁ to R ₃ may each independently be selected from the group consisting of hydrogen, halogen, C ₁ -C ₆ alkyl, and C ₁ -C ₆ alkoxy, and R ₄ is hydrogen or C It may be ₁ -C ₆ alkyl, but more preferably, in Formula 1, R ₁ and R ₂ may each independently be any one selected from the group consisting of hydrogen, halogen, and -O-CH ₃ , and the formula In 1, R ₃ and R ₄ may be hydrogen.

In Formula 1, R ₅ may be any one selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl, and C ₆ -C ₁₀ aryl, but more preferably R ₅ is hydrogen, C ₁ -C ₃ alkyl. , substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-10 arylalkyl.

Indirubin represented by Formula 1 is 5-methoxyl indirubin-3'-oxime, indirubin-3'-oxime, 6-bromoindirubin-3'-oxime, 5,6-dichloro indirubin, 5 , 6-dichloro indirubin-3'-oxime, 5,6-dichloro indirubin-3'-methoxime, 5,6-dichloro indirubin-3'-propyl oxime, 6-chloro-5-nitro indirubin, 6-chloro-5-nitroindirubin-3'-oxime, 6-chloroindirubin-3'-methoxime, 5-chloroindirubin-3'-methoxime, 5-bromoindirubin-3'-oxime , 5-bromoindirubin-3'-methoxime, 5-bromoindirubin-3'-ethyloxime, 5,6-dichloroindirubin-3'-oximepropyloxime and 6-chloroindirubin-3' -It may be any one selected from the group consisting of benzyloxime.

Most preferably, Formula 1 may be any one selected from the following Formulas 1-1 to 1-4, but is not limited thereto, and can significantly prevent and treat hair loss and hair growth through combined use with a metabolic activator. Alternatively, it may show an improvement effect, but most preferably, an indirubin derivative represented by Formula 1-1 prevents hair loss when administered in combination with a metabolic activator, not only promotes hair growth or hair growth, but also promotes the proliferation of skin cells. By increasing and inducing, a significant effect beyond the synergistic effect can be achieved compared to single administration.

[Formula 1-1]

[Formula 1-2]

[Formula 1-3]

[Formula 1-4]

The metabolic activator may be a PKM2 (pyruvate kinase M2) activator or an MPC (Mitochondrial pyruvate carrier) inhibitor, and the PKM2 (pyruvate kinase M2) activator refers to any substance that can promote the expression or activity of PKM2. , but is not particularly limited thereto, preferably 6-[(3-aminophenyl)methyl]-4,6-dihydro-4-methyl-2-(methylsulfinyl)-5H-thieno[2',3':4,5] pyrrolo[2,3-d]pyridazin-5-one(TEPP-46, ML265), 3-(4-(2,3-Dihydrobenzo[b][1,4]dioxin-6-ylsulfonyl)-1,4 -diazepan-1-ylsulfonyl)aniline (DASA-58) may be any one selected from the group consisting of serine, and more preferably 6-[(3-aminophenyl)methyl]-4,6-dihydro It may be -4-methyl-2-(methylsulfinyl)-5H-thieno[2',3':4,5]pyrrolo[2,3-d]pyridazin-5-one (TEPP-46, ML265).

In addition, the MPC (Mitochondrial pyruvate carrier) inhibitor refers to any substance that inhibits glucose or pyruvate through mitochondrial pyruvate carrier (MPC) inhibition, but is not particularly limited thereto, (E)-2-Cyano- It may be 3-(1-phenyl-1H-indol-3-yl)acrylic acid (UK5099).

In the present invention, the term 'hair loss' refers to a state in which there is no hair in areas where hair should normally exist, regardless of the cause, for example, male pattern baldness, female pattern baldness, alopecia areata, telogen effluvium, stress-related. It may be hair loss or hair loss caused by chemotherapy drugs. The composition of the present invention promotes proliferation of dermal papilla cells and growth of hair length, and therefore may be particularly effective in the treatment of male pattern baldness.

Preferably, the indirubin derivative is represented by Formulas 1-1 to 1-4, which shows little toxicity to cells even when treated with cells for a long period of time, and has a significant increase in effect when treated with a metabolic activator. It may be any one or more selected from the group consisting of, and most preferably, it may be an indirubin derivative represented by Formula 1-2. This is because through in vitro experiments, treatment with an indirubin derivative of Chemical Formula 1-2 and a metabolic activator was observed to increase the length of vibrissa by more than 75 to 95% compared to the conventional hair loss treatment agent (minoxidil). Because it has been done.

Therefore, the composition containing an indirubin derivative and a metabolic activator according to the present invention can be used as a pharmaceutical composition and cosmetic composition that prevents hair loss and promotes hair growth by preventing hair follicles from entering the telogen phase and maintaining the growth phase. You can.

In addition, the composition according to the present invention did not show inflammation or other pathological findings and was confirmed to be stable in cytotoxicity tests, and based on this, it can be used as an improvement or treatment for hair loss.

The indirubin derivative represented by Formula 1 according to the present invention can be used not only as a free substance, but also as a pharmaceutically acceptable salt, pharmaceutically acceptable solvate, pharmaceutically acceptable polymorph, or pharmaceutically acceptable prodrug. can be provided. The salt of the indirubin derivative is not particularly limited as long as it is in a form that can be incorporated into medicines or cosmetics. It contains inorganic salts or organic salts and may be acidic salts or alkaline salts. In particular, in the case of cationic salts, alkali metal salts such as sodium salts and potassium salts, alkaline earth metal salts such as calcium salts, magnesium salts, and barium salts; Basic amino acid salts such as arginine and lysine; Ammonium salts such as ammonium salt and tricyclohexylammonium salt; It may be various alkanol amine salts such as monoethanolamine salt, diethanolamine salt, triethanolamine salt, monoisopropanolamine salt, diisopropanolamine salt, and triisopropanolamine salt. Preferably, the salt is an alkali metal salt, and more preferably, it may be a tetrasodium salt.

The mixing ratio of the indirubin derivative and the metabolic activator is not particularly limited, but may be included at a molar ratio of 1:0.1 to 10, preferably 1:0.1 to 5, and more preferably 1:0.1 to 5. It may be a molar ratio of 0.1 to 3. If it is outside the above range, it is undesirable because the hair loss prevention and hair promotion effects are reduced.

The term 'comprising an active ingredient' in the present invention means containing an active ingredient in an amount sufficient to prevent hair loss or promote hair growth in the present invention.

As used herein, the term 'prevention' refers to any action that inhibits or delays hair loss or hair damage by administering the composition according to the present invention, and 'treatment' refers to improvement of hair loss symptoms by administering the pharmaceutical composition. It means any action that becomes or changes beneficially. The term 'improvement' in the present invention means any act of reducing at least the degree of symptoms, for example, parameters related to the hair loss or hair damage condition treated by administration of the composition containing the indirubin derivative and the metabolic activator of the present invention. means.

The pharmaceutical composition of the present invention can be administered parenterally or orally according to the desired method, and preferably may be parenteral administration or topical administration through application. The dosage range varies depending on the patient's weight, age, gender, health condition, diet, administration time, administration method, excretion rate, and severity of the disease. In addition, the therapeutically effective amount of the composition may vary depending on the administration method, target area, and patient's condition, and when used in the human body, the dosage should be determined as an appropriate amount considering safety and efficiency.

As used herein, “subject” refers to an object for which the disease is to be prevented or treated, and preferably includes humans and animals.

Since the active ingredient of the present invention has excellent stability to the living body as measured in Experimental Example 2, the upper limit of the dosage is not particularly limited, but the preferred dosage of the composition of the present invention is 0.01 to 1000 mL/kg per day. Preferably, it can be administered at 0.1 to 100 ml/kg, and more preferably at 1 to 10 ml/kg. Administration may be administered once a day, or may be administered several times. The above dosage does not limit the scope of the present invention in any way.

The pharmaceutical composition of the present invention can be formulated into oral dosage forms such as powders, granules, tablets, soft or hard capsules, suspensions, emulsions, syrups, and aerosols, skin external preparations such as ointments and creams, suppositories, injections, and the like, respectively, according to conventional methods. It can be formulated and used in any form suitable for pharmaceutical preparations, including sterile injectable solutions.

It may additionally include excipients such as fillers, extenders, binders, wetting agents, disintegrants, surfactants, and diluents commonly used for the formulation. For example, excipients that may be included in the pharmaceutical composition of the present invention include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate. , cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate, and mineral oil, but are not limited thereto. Additionally, in addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. As a base for suppositories, witepsol, macrogol, tween 61, cacao, laurel, glycerogelatin, etc. can be used.

The composition according to the present invention may have a dosage form that can be administered by a method such as directly applying or spraying to the skin or wound, for example, a cream, lotion, ointment, aerosol, gel, or pack. Methods for mixing ingredients or formulations suitable for each dosage form are well known in the art. When preparing these preparations, those skilled in the art can appropriately select and use various mixing ingredients used in preparing conventional external preparations.

In the case of ointments, creams, gels, lotions, etc., such ingredients include white petrolatum, yellow petrolatum, lanolin, bleached beeswax, cetanol, stearyl alcohol, stearic acid, hydrogenated oil, gelling hydrocarbons, polyethylene glycol, and liquid paraffin. , squalane, etc., oleic acid, isopropyl myrstate, glycerin triisooctanoate, crotamiton, diethyl sebacate, diisopropyl adipate, hexyl laurate, fatty acids, fatty acid esters, fatty alcohols, vegetable oils. There are solvents and solubilizers such as tocopherol derivatives, antioxidants such as L-ascorbic acid, dibutylhydroxytoluene, butylhydroxyanisole, preservatives such as parahydroxybenzoic acid ester, glycerin, propylene glycol, and sodium hyaluronate. Moisturizers such as polyoxyethylene derivatives, glycerin fatty acid ester, sucrose fatty acid ester, sorbitan fatty acid ester, propylene glycol fatty acid ester, surfactants such as lecithin, carboxyvinyl polymer, xanthan gum, carboxymethyl cellulose, carboxymethyl cellulose sodium salt, There are thickeners such as hydroxypropylcellulose and hydroxypropylmethylcellulose.

In the case of aerosol preparations, in addition to the above ingredients used in the preparation of ointments, creams, gels, suspensions, emulsions, solutions, lotions, etc., various stabilizers, buffers, corrigants, suspending agents, emulsifiers, fragrances, and preservatives are added. , solubility aids, and other suitable additives can be added.

The present invention relates to a cosmetic composition for alleviating, suppressing or improving hair loss containing an indirubin derivative represented by Formula 1 and a metabolic activator as active ingredients. The specific description of the cosmetic composition is the same as the description of the pharmaceutical composition for preventing or treating hair loss containing the indirubin derivative represented by Chemical Formula 1 and the metabolic activator as active ingredients, so please refer to that section for detailed information. .

Ingredients included in the cosmetic composition of the present invention include ingredients commonly used in cosmetic compositions in addition to the indirubin derivative and metabolic activator as active ingredients, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and fragrances. It may include common auxiliaries and carriers such as:

The cosmetic composition of the present invention can be prepared in any formulation commonly prepared in the art, for example, solution, suspension, emulsion, paste, gel, shampoo, rinse, conditioner, lotion, lotion, cream, serum. , essence, gel, pack, cleanser, soap, spray, powder, oil, powder foundation, emulsion foundation, and wax foundation, but is not limited thereto, and specifically hair tonic, hair conditioner, and 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. Any product selected from the group consisting of hair oil, hair dryer, hair preservative treatment, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, hair mousse, and hair spray. It may be a single hair formulation.

The present invention relates to a food composition for preventing or improving hair loss containing an indirubin derivative represented by Formula 1 and a metabolic activator as active ingredients. The specific description of the food composition is the same as the description of the pharmaceutical composition for preventing or treating hair loss containing the indirubin derivative represented by Chemical Formula 1 and the metabolic activator as active ingredients, so please refer to that section for detailed information. .

The food composition according to the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, gum, ice cream, vitamin complexes, and health supplements. etc.

The food composition of the present invention may contain not only the above active ingredients but also ingredients commonly added during food production, including, for example, proteins, carbohydrates, fats, nutrients, seasonings, and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose, etc.; Disaccharides such as maltose, sucrose, oligosaccharides, etc.; and polysaccharides, such as common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents (thaumatin, stevia extract (e.g., rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is manufactured as a drink or beverage, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and various plant extracts may be additionally included in addition to the boratin of the present invention.

The present invention provides a health functional food containing a food composition for preventing or improving hair loss containing an active ingredient. Health functional foods are foods manufactured by adding the active ingredients of indirubin derivatives and metabolic activators represented by Chemical Formula 1 to food ingredients such as beverages, teas, spices, gum, and confectionery, or by encapsulating, powdering, or suspending them. , which means that ingesting it will bring about certain health effects, but unlike regular drugs, it has the advantage of not having any side effects that may occur when taking the drug for a long time since it is made from food. The health functional food of the present invention obtained in this way is very useful because it can be consumed on a daily basis. The amount of active ingredients added in such health functional foods cannot be uniformly specified as it depends on the type of health functional food being targeted. However, it can be added within a range that does not damage the original taste of the food, and is usually 0.01% for the target food. to 50% by weight, preferably 0.1 to 20% by weight. In addition, in the case of health functional foods in the form of pills, granules, tablets, or capsules, it is usually added in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule, or beverage.

In addition, the present invention provides a composition for promoting the growth of dermal papilla cells or hair follicle cells isolated from mammalian skin, comprising an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients.

The specific description of the composition is the same as the description of the pharmaceutical composition for preventing or treating hair loss containing the indirubin derivative represented by Formula 1 and a metabolic activator as active ingredients, so refer to that section for detailed information.

The composition of the present invention can be used as a composition that can promote the proliferation of dermal papilla cells or hair follicle cells in vitro, and can especially be used as a medium composition that can culture the dermal papilla cells and hair follicle cells in vitro.

The mammal may be any one selected from the group consisting of humans, cattle, goats, and sheep.

The present inventors provide a composition that can efficiently and rapidly mass-produce dermal papilla cells or hair follicle cells in vitro in relation to hair loss or hair growth promotion, and the composition only efficiently causes the growth of dermal papilla cells or hair follicle cells. In addition, it was discovered that large amounts of non-toxic dermal papilla cells and hair follicle cells could be produced at high yield.

Hereinafter, the present invention will be described in more detail through examples, etc., but the scope and content of the present invention should not be construed as being reduced or limited by the examples below. In addition, based on the disclosure of the present invention, including the following examples, it is clear that a person skilled in the art can easily implement the present invention for which no specific experimental results are presented, and the patent to which such variations and modifications are attached It is natural that it falls within the scope of the claims.

In addition, the experimental results presented below describe only the representative experimental results of the examples and comparative examples, and the effects of each embodiment of the present invention that are not explicitly presented below will be described in detail in the corresponding section.

Preparation Example 1. Synthesis of 5,6-dichloroindirubin-3'-methoxime (KY19382).

① Intermediate 5',6'-dichloro-[2,3'-biindolinylidene]-2'3-dione (5',6'-dichloro-[2,3'-biindolinylidene]-2' ,3-dione) synthesis

Add 5,6-dichloroisatin (500 mg, 2.32 mmol) to a 250 ㎖ round bottom flask, dissolve in methanol (MeOH) (92.80 ㎖), and then add indoxyl acetate ( 405.48 mg, 2.315 mmol) and sodium carbonate (Na ₂ CO ₃ ) (637.83 mg, 6.02 mmol) were added and stirred at 65°C for 12 hours. The completion of the reaction was confirmed using TLC (Rf = 0.4, ethyl acetate/hexane = 1/2 (v/v)), and the product was cooled in ice until crystal lumps formed. If crystals form, filter them to remove the solvent, then discard the filtrate and wash the product several times with solvent (ethanol/water = 1/1 (v/v)). The product was filtered, dried in a vacuum pump and used in the next step without further purification.

② Synthesis of KY19382 (Formula 1-2)

5',6'-dichloro-[2,3'-biindolinylidene]-2'3-dione (5',6'-dichloro-[2,3'-biindolinylidene) in a 100 mL round bottom flask. ]-2',3-dione) (600 mg, 1.81 mmol) was added and dissolved in pyridine (151 ml), then H ₂ NOCH ₃ ·HCl (3026.4 mg, 36.24 mmol) was added, then 120 Stir at ℃ for 12 hours. Confirm the completion of the reaction using TLC (R _f = 0.4, ethyl acetate/hexane = 1/1 (v/v)) and lower the temperature of the reaction solution to room temperature. After evaporating all of the pyridine solvent in the product, water and ethyl acetate were added and the product was dissolved using ultrasound for 30 minutes, then extracted twice with ethyl acetate and washed with a saturated solution of sodium bicarbonate (NaHCO ₃ ). The extracted solution was dehydrated with anhydrous magnesium sulfate, the solvent was evaporated, and it was recrystallized using methanol and hexane. When the product is dried in a vacuum pump, red solid KY19382 (326 mg) can be obtained with a yield of 47.94%. ^1H NMR (400 MHz, DMSO- d6 ) δ 11.36 (s, 2H), 8.80 (s, 1H), 8.08 (d, 1H, J = 7.7 Hz), 7.46 - 7.41 (m, 2H), 7.07 - 6.99 (m, 2H), 4.38 (s, 3H).

Preparation Example 2. Synthesis of 5-methoxyl indirubin-3'-oxime

① Intermediate 5'-methoxy-[2,3'-biindolinylidene]-2',3-dione (5'-methoxy-[2,3'-biindolinylidene]-2',3-dione) synthesis of

Add 5-methoxyisatin (1000 mg, 5.65 mmol) to a 250 mL round bottom flask, dissolve in methanol (MeOH) (225 mL), and then add indoxyl acetate (989 mg, 5.65 mmol). mmol) and sodium carbonate (Na ₂ CO ₃ ) (1496 mg, 14.11 mmol) were added and stirred at 65°C for 12 hours. The completion of the reaction was confirmed using TLC (Rf = 0.4, ethyl acetate/hexane = 1/2 (v/v)), and the product was cooled in ice until crystal lumps formed. If crystals form, filter them to remove the solvent, then discard the filtrate and wash the product several times with solvent (ethanol/water = 1/1 (v/v)). The product was filtered, dried in a vacuum pump and used in the next step without further purification.

② Synthesis of 5-methoxyl indirubin-3'-oxime (Formula 1-1)

5'-methoxy-[2,3'-biindolinylidene]-2',3-dione (5'-methoxy-[2,3'-biindolinylidene]-2', 3-dione) (670 mg, 2.29 mmol) was added and dissolved in pyridine (27 ml), then H ₂ NOH·HCl (3186 mg, 45.85 mmol) was added, and stirred at 120°C for 12 hours. do. The completion of the reaction is confirmed using TLC (R _f = 0.5, ethyl acetate/hexane = 1/1 (v/v)), and the temperature of the reaction solution is lowered to room temperature. After evaporating all of the pyridine solvent in the product, water and ethyl acetate were added and the product was dissolved using ultrasound for 30 minutes, then extracted twice with ethyl acetate and washed with a saturated solution of sodium bicarbonate (NaHCO ₃ ). After dehydrating the extracted solution with anhydrous magnesium sulfate, the solvent is evaporated and recrystallized using methanol and hexane. When the product is dried in a vacuum pump, red solid 5-methoxyl indirubin-3'-oxime (Formula 1-1) (420 mg) can be obtained with a yield of 59%. ^1H NMR (400 MHz, DMSO- d6 ) δ 13.52 (s, 1H), 11.79 (s, 1H), 10.54 (s, 1H), 8.35 (d, J = 1.5 Hz, 1H), 8.26 (d, J = 7.6 Hz, 1H), 7.41 (d, J = 2.8 Hz, 2H), 7.08 - 7.00 (m, 1H), 6.82 - 6.71 (m, 2H), 3.78 (s, 3H).

Preparation Examples 3 and 4. Preparation of 5,6-dichloroindirubin-3'-oxime propyloxime and 6-chloroindirubin-3'-benzyloxime

5,6-dichloroindirubin-3'-propyloxime and 6-chloroindirubin-3'-benzyloxime are 5,6-dichloroindirubin-3'-methoxime or 5-methoxyl indirubin-3'. -It was synthesized in the same way as oxime, and was dissolved in dimethyl sulfide (DMSO) and applied to the experiment.

Examples and comparative examples. Mixture of indirubin derivatives and metabolic activators

As shown in Table 1 below, a mixture of indirubin derivatives and a metabolic activator was prepared. The metabolic activator used in the present invention is 6-[(3-aminophenyl)methyl]-4,6-dihydro-4-methyl-2-(methylsulfinyl)-5H-thieno[2',3':4,5]pyrrolo[ 2,3-d]pyridazin-5-one(TEPP-46, ML265), 3-(4-(2,3-Dihydrobenzo[b][1,4]dioxin-6-ylsulfonyl)-1,4-diazepan -1-ylsulfonyl)aniline (DASA-58) and (E)-2-Cyano-3-(1-phenyl-1H-indol-3-yl)acrylic acid (UK5099) and serine were obtained from Sigma Aldrich (St. Louis, USA) and used.

At this time, 150 μM of minoxidil (MNX), a commercially available drug, was used as a positive control, and DMSO treated instead of the mixture was used as a negative control.

division	Indirubin derivatives KY19382	metabolic activator
		type	content
Example 1	0.5 μM	TEPP-46	0.1 μM
Example 2	0.5 μM	DASA-58	0.1 μM
Example 3	0.5 μM	Serine	1mM
Example 4	0.5 μM	UK-5099	0.1 μM
Comparative Example 1	0.5 μM	-	-
Comparative Example 2	-	TEPP-46	0.1 μM
Comparative Example 3	-	DASA-58	0.1 μM
Comparative Example 4	-	Serine	0.1 μM
Comparative Example 5	-	Serine	1 μM
Comparative Example 6	-	Serine	10 μM
Comparative Example 7	-	Serine	0.1mM
Comparative Example 8	-	Serine	1mM
Comparative Example 9	-	UK-5099	0.1 μM
Comparative Example 10	-	Dichloro-acetate	0.1 μM
Comparative Example 11	0.5 μM	Dichloro-acetate	0.1 μM

Examples 1 to 4 are a mixture of an indirubin derivative represented by Formula 1-2 and a metabolic activator, and Comparative Examples 1 to 9 are a group administered only an indirubin derivative represented by Formula 1-2 or a metabolic activator. Comparative Examples 10 and 11 contain dichloroacetate, a PDK activator, not a PKM2 activator, alone or mixed with an indirubin derivative among metabolic activators.

Experimental Example 1. Measurement of hair growth efficacy in ex vivo

Male mice aged 6 to 8 weeks were sacrificed, and tissues around the mouth were isolated. The separated tissue was placed in 70% ethanol and then washed with PBS (phosphate) containing 1% penicillin (100IU/ml)/streptomycin (100㎍/ml) (P/S, GIBCO, Grand Island, NY, USA). -buffered saline)pH 7.4, SIGMA, St. Louis, MO, USA) to prepare an emulsion. Using a microscope, vibrissa follicles in the anagen state were separated one by one. The isolated vibrissa follicles were dispensed into each well of a 24-well plate and then treated with medium containing the examples or comparative examples (DMEM serum free 1% P/S + 12.5 μg/ml gentamycin). The plate was cultured at 37°C and 5% CO ₂ and observed through a microscope after 6 days. The medium was changed once every two days. The hair follicle morphology was photographed under a microscope (SMZ745T, Nikon), and the hair follicle length was measured using image J. The average value of hair follicle length change was calculated and compared with the average length of the negative control group or single treatment group to measure the degree of growth. The results obtained from the experiment were recorded as the mean and its standard error of the mean (SEM), and significance was determined using the Student's T-test analysis method.

Figure 1 is a micrograph taken after culturing Vibrissa hair follicles treated with indirubin derivatives or metabolic activators (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) alone for 6 days, and Figure 2 is This is a photomicrograph taken after culturing Vibrissa hair follicles treated with a mixture of indirubin derivatives and metabolic activators (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) for 6 days.

Figure 3 shows the length change of each Vibrissa hair follicle treated alone or in combination with an indirubin derivative and a metabolic activator (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) and cultured for 6 days. This is the graph shown (* P < 0.05, ** P < 0.005, *** P < 0.0005, NS: not significant vs. VEH or single treat).

Figure 4 is a micrograph (top) of Vibrissa hair follicles taken in the group administered with various concentrations of serine alone and a graph (bottom) showing the length change (* P < 0.05, ** P < 0.005, *** P < 0.0005, NS: not significant vs. VEH).

division	* relative vibrissa length	S.E.M.	Comparison to VEH	Comparison to single-treatment
Negative control group (VEH)	100	8.53575	-	-
Positive control group (MNX)	145.1296	7.973598	0.002173	-
Example 1	283.7517	26.28443	1.47E-06	0.025156
Example 2	254.5192	18.40625	1.66E-06	0.018985
Example 3	280.0675	29.58538	1.29E-06	0.00805
Example 4	269.6002	17.42833	8.63E-08	0.021875
Comparative Example 1	179.1396	8.083833	6.08E-06	-
Comparative Example 2	202.3379	14.27015	1.21E-05	-
Comparative Example 3	193.872	12.97311	1.13E-05	-
Comparative Example 4	116.3255	12.43135	0.369696	-
Comparative Example 5	117.0565	11.28889	0.327976	-
Comparative Example 6	128.6062	17.73216	0.204813	-
Comparative Example 7	156.291	19.96604	0.027517	-
Comparative Example 8	163.103	19.61302	0.004953	-
Comparative Example 9	202.1371	18.6263	0.000218	-
Comparative Example 10	160.6009	15.02112	0.000758	-
Comparative Example 11	194.1432	10.99876	9.19E-06	0.12758

*) Relative vibrissa length change value based on VEH

As shown in Figures 1 to 4 and Table 2, Examples 1 to 4, which are a mixture of an indirubin derivative represented by Formula 1-2 and a metabolic activator, have a ratio of 75 to 95% or more than minoxidil (MNX), a conventional hair loss treatment agent. It showed an effect of increasing the length of vibrissa, and the length of vibrissa increased compared to Comparative Examples 1 to 9 (single-treatment) in which the indirubin derivative or metabolic activator represented by Formula 1-2 was administered alone. It was confirmed that was 20% to 40% or more.

Considering that Comparative Examples 1 to 9, in which the indirubin derivative or metabolic activator represented by Formula 1-2 was administered alone, produced an increase effect of only about 1.25 times that of minoxidil (MNX), a conventional hair loss treatment, It can be seen that Examples 1 to 4 achieve significantly more significant effects than the simple synergistic effect of Comparative Examples 1 to 9.

Therefore, it can be seen that the mixed composition prepared in Examples 1 to 4 of the present invention can be used as a composition useful for preventing hair loss and promoting hair growth.

Experimental Example 2. Toxicity Analysis

Human dermal papilla cells were fixed at a rate of 2 × 10 ⁵ per well of a 6-well plate, and when the cells were stably attached, they were treated with the drugs of Examples or Comparative Examples. . After 24 hours, luminescence values were measured using a FLUOstar OPTIMA luminometer (BMG Labtech, Offenburg, Germany) using the Cell Titer-Glo Luminescent Cell Viability Assay kit (Promega, Madison, USA).

The results obtained from the experiment were recorded as the mean and its standard error of the mean (SEM), and significance was determined using the Student's T-test analysis method.

Figure 5 shows the cell viability of human dermal papilla cells treated with indirubin derivatives and metabolic activators (TEPP-46, DASA-58, Serine, UK-5099, Dichloroacetate) alone or in combination. This is the graph shown (* P < 0.05, ** P < 0.005, *** P < 0.0005 vs VEH).

As shown in Figure 5, the indirubin derivative represented by Formula 1-2 and the metabolic activator did not show cytotoxicity either alone or in combination. That is, in Examples 1 to 4 and Comparative Examples 1 to 12, it was confirmed that the cell viability was more than 100%, confirming that the cell stability was very excellent. However, in the case of minoxidil (MNX), it was confirmed to be 97%, the only one below 100%.

Experimental Example 3. Animal experiment

C57BL/6N mice were used as experimental animals considering ease of experimentation. The experimental animals used were 7-week-old male C57BL/6N normal mice weighing approximately 25g purchased from Orient Bio Co., Ltd. Five animals were assigned to each experimental group. In C57BL/6N mice, all hair follicles enter the anagen phase of the hair growth cycle from birth and begin to grow hair. After about 3 weeks, the mouse transitions to the telogen phase and immediately enters the second growth phase. It is known that all hair follicles transition into the telogen phase around 6-7 weeks, so this telogen phase is appropriate for evaluating the efficacy of only the experimental substance since the influence of the test animal's own hair growth factors is excluded.

To review the ethical and scientific validity and efficient management of animal experiments, this study was conducted with approval from the Institutional Animal Care and Use Committee (IACUC) of Yonsei University.

The hair on the back of a 7-week-old C57BL/6N mouse was removed using an animal clipper. The experiment was conducted by dividing the animals into 5 groups with 5 animals per group. Except for the normal group, 150 μl of the drug was applied to the back skin once a day for 28 days. Drugs were administered, and 4 weeks later, they were euthanized using carbon dioxide and photographs were taken. To compare the degree of hair growth, hair growth during the anagen phase was observed after 4 weeks by measuring the weight of the regrown hair. Among all areas of hair loss, the weight of the area where hair grew during the anagen phase was measured.

The results obtained from the experiment were recorded as the mean and its standard error of the mean (SEM), and significance was determined using the Student's T-test analysis method.

Animal models from 5 groups

Normal group (VHE): 150 μl PBS was applied to the back skin once a day for 28 days.

Experimental group (KY+TE): 150 μl of a mixture of KY19382 (0.5mM) and TEPP-46 (0.5mM) was applied to the back skin once a day for 28 days.

First comparison group (TEPP46): 150 μl of TEPP-46 (0.5 mM) was applied to the back skin once a day for 28 days.

Second comparison group (KY19382): 150 μl of KY19382 (0.5 mM) was applied to the back skin once a day for 28 days.

Third comparison group (MNX): 150 μl of minoxidil (MNX) (150 mM) was applied to the back skin once a day for 28 days.

Figure 6 is a photograph taken of each group treated with indirubin derivative and metabolic activator (TEPP-46) alone or in combination, and Figure 7 is a photograph of each group treated with indirubin derivative and metabolic activator (TEPP-46) alone or in combination. This is a graph showing the hair growth values measured from each treated group (* P < 0.05, ** P < 0.005, *** P < 0.0005, NS: not significant vs VEH or single treat).

As shown in Figures 6 and 7, hair growth was observed through the weight of regrown hair, and it was confirmed that the degree of hair growth in the experimental group significantly increased compared to the MNX group after 4 weeks. In the experimental group treated with a mixture of indirubin derivative and metabolic activator (TEPP-46), the degree of hair growth significantly increased compared to the first and second comparison groups.

As a result of analyzing the photo in FIG. 6 for visual observation of the wool, in the experimental group treated with a mixture of indirubin derivative and metabolic activator (TEPP-46) 4 weeks after the experiment, the ratio of the area where hair grew out of the total hair loss area on the back was It was confirmed that there was a significant increase compared to the first to third comparison groups. In addition, in the experimental group treated with a mixture of an indirubin derivative represented by Formula 1-2 and a metabolic activator (TEPP-46), no inflammation or other pathological findings were confirmed, and hair grew without gaps in the entire hair loss area. The amount and length of hair increased more than before hair removal.

That is, the composition combining the indirubin derivative (KY19382) and the metabolic activator (TEPP-46) of the present invention was compared and reviewed for hair loss improvement effect with MNX, an existing hair loss improvement agent, and as a result, the indirubin derivative (KY19382) and the metabolic activator were compared. It was confirmed that the composition mixed with the agent (TEPP-46) promoted hair growth during the growth period and showed significantly significant hair growth efficacy. In addition, the mixed composition of indirubin derivative (KY19382) and metabolic activator (TEPP-46) did not show inflammation or other pathological findings and was confirmed to be stable in cytotoxicity tests. Based on this, it was confirmed as an improvement or treatment for hair loss. You can see that it is available.

Formulation Example 1: Cream-type cosmetic composition for preventing or improving hair loss

In this formulation example, an attempt was made to prepare a cream-type cosmetic composition for preventing or improving hair loss containing a mixed composition of an indirubin derivative of Formula 1-2 and a metabolic activator (TEPP-46) as an active ingredient. Components including the composition of Example 1 were mixed according to the following amounts to prepare a cream-type cosmetic composition for preventing or improving hair loss.

4.0% by weight of the composition of Example 1

Glycerin 2.0% by weight

Paraben 0.2% by weight

Allantoin 2.0% by weight

Betaine 3.0% by weight

Sodium hyaluronate 1.0% by weight

Tocopherol acetate 5.0% by weight

Shea butter 2.0% by weight

Trehalose 1.0% by weight

Weight percent of preservatives and fragrances

Purified water to 100% by weight

Total 100% by weight

Formulation Example 2: Pharmaceutical composition (ointment) for preventing or treating hair loss

In this formulation example, an attempt was made to prepare a pharmaceutical composition (ointment) for preventing or treating hair loss containing a mixed composition of an indirubin derivative of Formula 1-2 and a metabolic activator (TEPP-46) as an active ingredient. Components including the composition of Example 1 were mixed according to the following amounts to prepare a pharmaceutical composition (ointment) for preventing or treating hair loss.

10% by weight of the composition of Example 1

Diethyl sebacate 8% by weight

Hard wax 5% by weight

Polyoxyethylene oleyl ether phosphate 6% by weight

% by weight of sodium benzoate

Vaseline to 100% by weight

Total 100% by weight

Formulation Example 3: Preparation of hair tonic

10.0% by weight of the composition of Example 1

Resorcinol 0.1% by weight

Menthol 0.05% by weight

Panthenol 0.2% by weight

Salicylic acid 0.1% by weight

Tocopheryl acetate 0.1% by weight

Pyridoxine hydrochloride 0.1% by weight

Castor oil 5.0% by weight

Pigment dosage

Spice Proper Amount

Ethanol dosage

Purified water remaining

Total 100.0% by weight

Formulation Example 4: Preparation of hair conditioner

2.5% by weight of the composition of Example 1

Cetanol 3.5% by weight

Self-emulsifying glycerin monostearate 1.5%

Propylene glycol 2.5%

Stearylmethylbenzyl ammonium chloride (25%) 7.0%

Methyl paraoxybenzoate 0.3%

Pigment dosage

Spice Proper Amount

Purified water remaining

Total 100.0%

Formulation Example 5: Preparation of hair lotion

5.0% by weight of the composition of Example 1

Resorcinol 2.0%

Menthol 2.0%

Panthenol 0.5%

Piroctonolamine 0.1%

Fragrance, coloring 0.5%

Purified water remaining

Total 100.0%

Formulation Example 6: Preparation of hair soap

0.1% by weight of the composition of Example 1

Titanium dioxide 0.2%

Polyethylene glycol 0.8%

Glycerin 0.5%

Ethylenediaminetetraacetic acid 0.05%

Sodium 1.0%

Pigment dosage

Soap scent appropriate amount

Toilet soap base (moisture 13%, parts by weight) remaining amount

Total 100.0%

Formulation Example 7: Application to milk

Milk 99.9% by weight

0.1% by weight of the composition of Example 1

Formulation Example 8: Preparation of beverage

10 mg of composition of Example 1

Calcium lactate 50 mg

Citric acid 5 mg

Nicotinamide 10 mg

Sodium riboflavin hydrochloride 3 mg

Pyridoxine hydrochloride 2 mg

Arginine 10 mg

Sucrose fatty acid ester 10 mg

200 ml of water

Claims (14)

1. Contains an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients,

   Pharmaceutical composition for preventing or treating hair loss, characterized in that the metabolic activator is a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate carrier) inhibitor:

   [Formula 1]

   

   In Formula 1,

   R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.
2. According to paragraph 1,

   In Formula 1,

   R ₁ to R ₃ are each independently selected from the group consisting of hydrogen, halogen, C ₁ -C ₆ alkyl, and C ₁ -C ₆ alkoxy,

   R ₄ is hydrogen or C ₁ -C ₆ alkyl,

   R ₅ is a pharmaceutical for preventing or treating hair loss, characterized in that any one selected from the group consisting of hydrogen, C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-10 arylalkyl. Composition.
3. According to paragraph 1,

   In Formula 1, R ₃ and R ₄ are hydrogen. A pharmaceutical composition for preventing or treating hair loss.
4. According to paragraph 3,

   In Formula 1, R ₁ and R ₂ are each independently selected from the group consisting of hydrogen, halogen, and -O-CH _3. A pharmaceutical composition for preventing or treating hair loss.
5. According to paragraph 1,

   In Formula 1, R ₅ is any one selected from the group consisting of hydrogen, C ₁ -C ₃ alkyl, benzyl group, and phenylpropyl group. A pharmaceutical composition for preventing or treating hair loss.
6. According to paragraph 1,

   Formula 1 is a pharmaceutical composition for preventing or treating hair loss, characterized in that any one selected from the following Formulas 1-1 to 1-4.

   [Formula 1-1]

   

   [Formula 1-2]

   

   [Formula 1-3]

   

   [Formula 1-4]

   
7. According to paragraph 1,

   The PKM2 (pyruvate kinase M2) activator is 6-[(3-aminophenyl)methyl]-4,6-dihydro-4-methyl-2-(methylsulfinyl)-5H-thieno[2',3':4,5 ]pyrrolo[2,3-d]pyridazin-5-one(TEPP-46, ML265), 3-(4-(2,3-Dihydrobenzo[b][1,4]dioxin-6-ylsulfonyl)-1, It may be any one selected from the group consisting of 4-diazepan-1-ylsulfonyl)aniline (DASA-58) and serine,

   The MPC (Mitochondrial pyruvate carrier) inhibitor is (E)-2-Cyano-3-(1-phenyl-1H-indol-3-yl)acrylic acid (UK5099). A pharmaceutical composition for preventing or treating hair loss. .
8. According to paragraph 1,

   A pharmaceutical composition for preventing or treating hair loss, characterized in that the indirubin derivative and the metabolic activator are contained in a molar ratio of 1:0.1 to 10.
9. Contains an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients,

   A cosmetic composition for preventing or improving hair loss, wherein the metabolic activator is a PKM2 (pyruvate kinase M2) activator or an MPC (Mitochondrial pyruvate carrier) inhibitor:

   [Formula 1]

   

   In Formula 1,

   R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.
10. According to clause 9,

    The composition is a cosmetic composition for preventing or improving hair loss, characterized in that it promotes the proliferation of dermal papilla cells or hair follicle cells in vitro.
11. According to clause 9,

    The cosmetic composition includes 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, and hair aerosol. , hair pack, hair nutrition pack, hair soap, hair cleansing foam, hair oil, hair dryer, hair preservation treatment, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dresser, hair lacquer, hair moisturizer, A cosmetic composition for preventing or improving hair loss, characterized in that it is a hair formulation selected from the group consisting of hair mousse and hair spray.
12. Contains an indirubin derivative represented by the following formula (1) and a metabolic activator as active ingredients,

    Food composition for preventing or improving hair loss, characterized in that the metabolic activator is a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate carrier) inhibitor:

    [Formula 1]

    

    In Formula 1,

    R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.
13. A method of treating hair loss comprising administering a therapeutically effective amount of an indirubin derivative represented by the following formula (1) and a metabolic activator to a hair loss patient,

    A method of treating hair loss, characterized in that the metabolic activator is a PKM2 (pyruvate kinase M2) activator or an MPC (Mitochondrial pyruvate carrier) inhibitor:

    [Formula 1]

    

    In Formula 1,

    R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.
14. For the use of indirubin derivatives and metabolic activators represented by the following formula (1) for the production of medicines for preventing or treating hair loss,

    Uses wherein the metabolic activator is a PKM2 (pyruvate kinase M2) activator or MPC (Mitochondrial pyruvate carrier) inhibitor:

    [Formula 1]

    

    In Formula 1,

    R ₁ to R ₅ are each independently hydrogen, halogen, hydroxy, nitro, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₆ -C ₁₀ aryl, and C _7-30 aryl. It is any one selected from the group consisting of alkyl.

Images