WO2019039755A2 - Composition for promoting hair restoration or hair growth comprising trimethyl cyclohexene compound as active ingredient - Google Patents

Abstract

The present invention relates to a composition for promoting hair restoration or hair growth comprising a trimethyl cyclohexene compound as an active ingredient. The composition according to the present invention has an effect of promoting hair dermal papilla cells and a hair follicle activating factor, thereby promoting hair sprouting or hair growth. Accordingly, the composition of the present specification may be utilized as a pharmaceutical composition or a cosmetic composition in various fields such as in hair loss treatment.

Description

Composition for promoting hair growth or hair growth comprising trimethylcyclohexene compound as an active ingredient

Disclosed herein is a composition for promoting hair growth or hair growth comprising a trimethyl cyclohexene compound as an active ingredient.

Human hair primarily acts as a defense function to protect the scalp from external stimuli such as ultraviolet rays and plays an aesthetic function expressing the external image of the individual. In modern society, however, hair loss is frequently caused by environmental factors such as environmental pollution, strong ultraviolet rays, stress, and hormonal imbalance. Unlike the past, which is confined to adult males, Therefore, the need for materials for prevention and treatment of these diseases has become urgent.

A hair follicle, which is a physiological organ in the body that produces hair, is an anagen phase in which hair is generated and generated hair is actively grown during postnatal development, a catagen phase in which hair is degenerated, The hair growth cycle is divided into the telogen phase, which is maintained until the hair loss, and the exogen phase, where hair loss occurs. Many studies have shown that hair follicle activity is caused by dermal papilla cells, and in particular, the proliferation and differentiation of dermal papilla cells are primarily involved in the progression of the hair growth cycle and hair formation. In the growth phase where the hair grows actively, the active growth and differentiation of the papillary cells occurs actively, the growth of the hair is stopped, and the cells are killed in the regenerator, the resting period and the hair removal period where the hair loss occurs.

Therefore, hair growth and hair loss are closely related to the proliferation and death of the dermal papilla cells. Therefore, prolonging the growth period or inhibiting apoptosis and shortening the period of regrowth, dormancy and hair loss by inducing proliferation of the dermal papilla cells will be a remedy . Cell division and migration in the vicinity of the dermal papilla is closely related to the growth of the hair. In the growing phase, hair is newly generated from the dermal papilla. Cells are activated by various cytokines and hormones, Of the growth rate.

In addition, growth factors such as HGF, IGF-1, and VEGF are most strongly expressed in the growth phase and decrease in the regressor and dormant period, thereby inducing the proliferation of the dermal papilla cells and maintaining the growth period, It is known to help.

Minoxidil and finasteride, which are currently approved by the FDA, suffer from side effects such as hair growth and lowering of sexual function, and the specific mechanism of action of minoxidil has not been clarified. Therefore, it is necessary to develop a biological material that promotes the proliferation of dermal papilla cells in hair follicles, increases the expression of growth factors, and has less side effects.

As described above, growth factors related to proliferation of dermal papilla cells and growth of hair are very important factors in normal hair maintenance, hair loss suppression and hair growth. The present inventors have completed the present invention by experimentally confirming that the trimethylcyclohexene compound promotes the growth of the papilla cells and increases the expression of growth factors such as IGF-1.

Accordingly, an object of the present invention is to provide a composition for promoting hair growth or hair growth, which comprises a trimethylcyclohexene compound as an active ingredient.

In one aspect, the present invention provides a composition for promoting hair growth or hair growth comprising a trimethyl cyclohexene compound as an active ingredient.

In one aspect, the composition for promoting hair growth or hair growth comprising the trimethylcyclohexene compound of the present invention as an active ingredient promotes the growth of the dermal papilla cells and the expression of growth factors, and for this purpose, A cosmetic composition, or an external preparation for skin.

Figure 1 shows the results of a proliferation and cytotoxicity assay of a composition comprising a trimethylcyclohexene compound (hereinafter referred to as PAC-6) according to an embodiment of the present invention and a positive control adiponectin (APN) .

Figure 2 is a graph showing the effect of a composition comprising a trimethylcyclohexene compound (hereinafter referred to as PAC-6) according to an embodiment of the present invention and a composition comprising adiponectin (APN) as a positive control, ) Were measured.

Term Definition

In this specification, when a part is referred to as " including " an element, it is to be understood that it may include other elements as well, without departing from the other elements unless specifically stated otherwise.

DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

Hereinafter, the present invention will be described in detail.

In an exemplary embodiment of the present invention, the present invention is a composition for promoting hair growth or hair growth, comprising a trimethylcyclohexene compound represented by the following formula (1) as an active ingredient.

[Chemical Formula 1]

The IUPAC name of the trimethylcyclohexene compound of Formula 1 is 3- (2,6,6-trimethyl-cyclohex-1-enyl) -acrylic acid 5-hydroxy-4-oxo-4H- 3- (2,6,6-Trimethyl-cyclohex-1-enyl) -propenoic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester.

The compound of Formula 1 is an off-white solid compound at room temperature.

The trimethylcyclohexene compound represented by the present invention includes isomers. The term " isomers " refers in particular to optical isomers (for example, essentially pure enantiomers, essentially pure diastereomers or mixtures thereof) as well as conformation isomers Position isomers (especially tautomers) or geometric isomers (e.g., cis-trans isomers), which are different from each other (i.e., isomers differing only in the angle of one or more chemical bonds).

The compound represented by Formula 1 is prepared by reacting a pyrenone derivative compound of Formula 2 and a cyclohexenyl ester compound of Formula 3 as shown in Reaction Scheme 1 below:

[Reaction Scheme 1]

(In the above Reaction Scheme 1, X is a halogen element and M is Li, Na or K)

Referring to Reaction Scheme 1, the compound of Formula 1 is prepared by a coupling reaction between a halogen element of a pyrazine compound and a metal of a cyclohexenyl ester compound.

Wherein X is a halogen element, wherein the halogen element may be F, Cl, Br, or I, preferably Cl. The compound of formula (2) can be used by purchasing a commercially available compound or by directly preparing it.

In the examples of the present invention, 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one was used in which X is Cl, which was obtained from 5-hydroxy-2- (hydroxymethyl) -4-one with thionyl chloride (SOCl ₂ ) and used directly.

In addition, the cyclohexenyl ester compound of formula (3) can be prepared by reacting 3- (2,6,6-trimethyl-cyclohex-2-enyl) propenoic acid, 3- (2,6,6-trimethyl-cyclohex-2-enyl) propenoic acid, 3- (2,6,6- (M < + >) bonded to the carboxyl group of the carboxyl group.

The cation (M ⁺ ) is one kind selected from the group consisting of Li ⁺ , Na ⁺ , and K ⁺ . (2,6,6-trimethyl-cyclohex-2-enyl) propenoic acid and Na < ⁺ > For example, 3- (2,6,6-trimethyl-cyclohex-2-enyl) -propenoic acid (3- (2,6,6- propenoic acid) and sodium hydroxide are dissolved together in methanol and ionized, followed by distillation of methanol.

In a preferred embodiment of the present invention, 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one is reacted with the compound of formula 2 and sodium 3- (2,6,6-trimethyl- (2, 6-trimethyl-cyclohex-2-enyl) propenoate) was prepared by ester linkage of the hydroxypyranone compound of formula (1).

At this time, the reaction is not particularly limited in the present invention and can be carried out under conditions that the halogen-metal bond reaction can be sufficiently performed.

The reaction temperature is, for example, 0.5 to 5 hours, preferably 1 to 3 hours at 50 to 250 ° C at the reflux temperature of the solvent.

The solvent may be any solvent capable of sufficiently dissolving the compounds of formulas (2) and (3). Examples of the solvent include N, N-dimethylformamide (DMF), tetrahydrofuran (DMSO), acetonitrile, dioxane, benzene, toluene, ether, methanol, hexane, cyclohexane, pyridine, N-methylpyrrolidone and combinations thereof, preferably DMF Lt; / RTI >

After the reaction, the solvent is distilled and subjected to a post-treatment such as usual washing, drying, and purification to obtain a highly pure compound.

When the trimethylcyclohexene compound according to the present invention is contained as an active ingredient, it promotes the growth of the papilla cells and promotes the expression of the growth factors IGF-1, VEGF, and HGF to promote hair growth.

In the present specification, in particular, in the test examples described later, when the effect of the composition is tested on dermal papilla cells derived from human rather than animals such as rats and the composition is applied to a human body as a cosmetic or pharmaceutical composition, It is evident that there is a promoting effect.

In one embodiment, the concentration of the active ingredient may be in the range of 1 to 100 [mu] M based on the total volume of the composition, and may be, for example, at least 10 [mu] M, at least 15, at least 20, at least 25, at least 30, , Or greater than or equal to 40 μM, and can be less than 90 μM, less than 80 μM, less than 70 μM, or less than 60 μM. In one embodiment herein, the concentration of the active ingredient is preferably 50 [mu] M.

If the concentration is less than 1 μM, the effect of promoting hair growth or hair growth may be insignificant. If the concentration is more than 100 μM, cytotoxicity may occur.

In one embodiment, the composition may be a pharmaceutical composition, a cosmetic composition, or an external preparation for skin.

The pharmaceutical composition for promoting hair growth or hair growth according to an embodiment of the present invention may be added with preservatives, stabilizers, wetting agents or emulsifying accelerators, pharmaceutical adjuvants such as salts and / or buffers for controlling osmotic pressure and other therapeutically useful substances And may be formulated into various oral or parenteral dosage forms according to conventional methods.

Examples of the oral administration agent include tablets, pills, hard and soft capsules, liquids, suspensions, emulsions, syrups, powders, powders, granules, granules and pellets, , Diluents (such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (such as silica, talc, stearic acid and magnesium or calcium salts thereof and polyethylene glycols) . The tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally mixed with starch, agar, alginic acid or its sodium salt Such as disintegrants, absorbents, coloring agents, flavoring agents, and sweetening agents. The tablets may be prepared by conventional mixing, granulating or coating methods. In addition, it may be a transdermal dosage form in the form of parenteral administration. For example, it may be in the form of injections, drops, ointments, lotions, gels, creams, sprays, suspensions, emulsions, suppositories, But is not limited thereto.

The pharmaceutical composition according to one embodiment of the present invention may be administered by parenteral, rectal, topical, transdermal, subcutaneous, and the like. The pharmaceutical composition according to one embodiment of the present invention may be administered topically, for example, to the scalp.

The dosage of the active ingredient is within the level of those skilled in the art, and the daily dose of the drug depends on various factors such as the degree of progress of the subject to be administered, the age of onset, age, health condition, Kg of the composition may be generally from 1 μg / kg to 100 mg / kg, for example, 0.1 mg / Kg to 20 mg / Kg, 0.5 mg / Kg to 20 mg / Kg, or 1 mg / kg to 20 mg / May be administered in a divided dose of 5 mg / kg to 10 mg / kg once or three times a day, and the dose is not limited to the scope of the present invention by any means.

The composition for promoting hair growth or hair growth according to an embodiment of the present invention may be a cosmetic composition, and the cosmetic composition may contain a cosmetically or dermatologically acceptable medium or base. It may be any formulation suitable for topical application, for example, as a solution, a gel, a solid, a paste anhydrous product, an emulsion obtained by dispersing an oil phase in water, a suspension, a microemulsion, a microcapsule, And in the form of non-ionic follicular dispersants, or in the form of creams, skins, lotions, powders, ointments, sprays or conical sticks. These compositions may be prepared according to conventional methods in the art. The composition according to the invention may also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant.

The cosmetic composition according to an embodiment of the present invention is not particularly limited in its formulation, and examples thereof include softening agents, convergent lotion, nutritional lotion, nutritional cream, massage cream, essence, eye cream, eye essence, Cream, cleansing foam, cleansing water, pack, powder, body lotion, body cream, body oil and body essence.

When the formulation of the present invention is a paste, cream or gel, animal fiber, plant fiber, 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.

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

When 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, linolenic derivatives or ethoxylated glycerol fatty acid esters.

The cosmetic composition according to an embodiment of the present invention may further contain ingredients included in the functional additive and the general cosmetic composition in addition to the above-described effective ingredients. The functional additives may include water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymeric polysaccharides, sphingolipids and seaweed extracts.

The cosmetic composition of the present invention may further contain, in addition to the above-described functional additive, components contained in a general cosmetic composition as required. Examples of the other ingredients that can be included in the composition include humectants, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbents, preservatives, bactericides, antioxidants, plant extracts, pH adjusters, alcohols, Accelerators, coolants, antiperspirants, purified water, and the like.

Further, the composition for promoting hair growth or hair growth according to an embodiment of the present invention is an external preparation for skin, and the external preparation for skin may be any kind applied to the outside of the skin, and various cosmetic formulations and medicines may be included therein have.

In one embodiment, the composition may further comprise vitamin D. The composition further includes vitamin D, thereby supplying nutrients to the hair follicle. The amount thereof is preferably 0.0001 to 10% by weight, particularly 0.01 to 1% by weight of the total composition.

In another exemplary embodiment of the present invention, the present invention provides a method for promoting hair growth or hair growth by administering an effective amount of a trimethyl cyclohexene compound represented by the following formula (1) to a subject in need thereof.

[Chemical Formula 1]

In another exemplary embodiment of the present invention, the present invention is a use of a trimethylcyclohexene compound represented by the following formula (1) for producing a composition for promoting hair growth or hair growth.

[Chemical Formula 1]

In another exemplary embodiment of the present invention, the present invention is a trimethylcyclohexene compound represented by the following formula (1) for promoting hair growth or hair growth.

[Chemical Formula 1]

Hereinafter, the present invention will be described more specifically with reference to the following examples. However, the following embodiments are provided for illustrative purposes only in order to facilitate understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

Manufacturing example : 3- (2,6,6- Trimethyl - Cyclohexane -One- Enil ) -Acrylic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester (3- (2,6,6- Trimethyl - cyclohex -One- enyl ) - propenoic  acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester

(2,6,6-trimethyl-cyclohex-1-enyl) -acrylic acid 5-hydroxy-4-oxo-4H-pyran-2 2-ylmethyl ester (3- (2,6,6-Trimethyl-cyclohex-1-enyl) -propenoic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester.

[Reaction Scheme 2]

The detailed procedure of the above reaction scheme 2 is as follows.

(0.35 mmol) of 5-hydroxy-2- (hydroxymethyl) -4H-pyran-4-one (0.35 mmol) was dissolved in 250 ml of N, N-dimethylformamide, cooled in an ice- 0.42 mol) was added dropwise over 30 minutes. After stirring at room temperature for 2 hours, the reaction solution was added to 2000 ml of ice water. The resulting solid was filtered, and a solid (filtrate) was dissolved in 1000 ml of ethyl acetate. Magnesium sulfate and activated charcoal were added, dried, discolored and filtered, and the filtrate was concentrated and nucleic acid was added to obtain crystals. And dried under vacuum to obtain 39.5 g (70%) of 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one as a reaction product as a yellow solid.

Subsequently, 5 g (0.026 mol) of 3- (2,6,6-trimethyl-cyclohex-2-enyl) -propenoic acid (3- (2,6,6- 1.3 g (0.031 mol) of sodium hydroxide was dissolved in 40 ml of methanol. The methanol was distilled off and the residue was dissolved in 70 ml of N, N-dimethylformamide to prepare the compound of Formula 3-1.

4.2 g (0.026 mol) of 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one of the above prepared formula 2-1 was added to the compound of the formula 3-1 thus prepared, Lt; / RTI > for 2 hours. The solvent was distilled off and the residue was dissolved in 300 ml of ethyl acetate. The ethyl acetate solution was washed with 5% hydrochloric acid and distilled water, dried over magnesium sulfate and activated carbon, and discolored. The insolubles were then filtered off and the filtrate was evaporated under reduced pressure to give 5.9 g (69% yield) of the reaction product as a off-white solid.

TLC (ethyl acetate: hexane = 1: 1) Rf = 0.43

¹ H NMR (DMSO-d 6,?): 9.22 (s, IH), 8.09 (s, IH), 6.75 , 5.50 (s, 1H), 5.02 (s, 2H), 2.45 (m, 2H), 2.00 (m, 2H), 1.42 0.80 (s, 3 H)

Example

Example 1: Proliferation and cytotoxicity analysis

Cell culture and differentiation

Human subcutaneous fat cells (hSCFs) and subcutaneous fat precursor cell differentiation medium were purchased from ZenBio Inc. (NC, USA) and cultured in a CO ₂ incubator with a humidity of 5%. To induce differentiation, human subcutaneous fat (hSCFs) were incubated with 10% fetal bovine serum (FBS, PAA, Pasching, Austria), 10 μg / ml insulin (Sigma-Aldrich, MO, USA), 0.5 mM 3-isobutyl- 1 μM dexamethasone (DEX, Sigma-Aldrich, MO, USA), 1-methylxanthine (IBMX; Sigma-Aldrich, St. Louis, (DMEM; Lonza, Md, USA) containing 1 μM troglitazone (Sigma-Aldrich, St. Louis, Mo., USA) The badge was replaced every other day.

Human breast milk (HDP) and HDP maintenance medium were purchased from Cefobio Co. (Seoul, Korea) and cultured in a CO ₂ incubator with a humidity of 5% according to the manufacturer's instructions. To determine the effect of adiponectin (APN), subcutaneous fat-containing culture medium and human-derived papillary (HDP) medium were mixed at 1: 0.25, 1: 0.5, and 1: Every other day.

Cell proliferation and cytotoxicity analysis

The viability of hSCFs was measured using the EZ-Cytox cell viability assay kit (MTT assay, Daeil Lab Service, South Korea) according to the manufacturer's instructions. In summary, hSCF was cultured for 7 days and treated with various concentrations of each chemical (adiponectin, hSCFs culture medium treated with PAC-6) for 24 and 72 hours, respectively. Then, EZ-Cytox solution (10 占 퐇) was added to each well and cultured at 37 占 폚 for 2 hours. Absorbance at 450 nm was measured using a spectrophotometer (Synergy H2, BioTek., VT, USA). All experiments were performed 3 times and data were expressed as absorbance.

As a result of the experiment, it was confirmed that the effect of the present invention on dermal papilla cell proliferation was similar to that of adiponectin, which is a positive control of the compound of the present invention, and that keratinocyte differentiation and hair generation can be promoted more effectively.

In addition, the cytotoxicity was also found to be equivalent to that of the normal control, and it was confirmed that it was not harmful to human body.

Example  2: Analysis of hair growth-related gene expressions

Real-time quantitative PCR (RT-qPCR)

Total RNA was extracted from the dermal papilla cells cultured in Example 1 using TRIzol reagent (Life Technologies, Carlsbad, Calif., USA) according to the manufacturer's instructions, and the RevertAid first strand cDNA synthesis kit (Thermo Scientific, Pittsburgh, PA, USA) was used to synthesize complementary DNAs (cDNAs). About 1 μg of cDNA sample and each TaqMan® probe (Life Technologies, Carlsbad, Calif., USA) were diluted in the reaction mixture of Quantitect Probe PCR kit (Qiagen, Valencia, CA, USA) Fast real-time PCR system) (Life Technologies, Carlsbad, Calif., USA). Each TaqMan® probe is: glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Part # 4352339E). All data are expressed as a multiple of change compared to the GAPDH control and were obtained from three independent experiments.

The expression level of IGF-1 (insulin-like growth factor), VEGF (vascular endothelial growth factor), HGF (hepatocyte growth factor) and hepatocyte growth factor 2, the expression of IGF-1 was 2-3 fold higher than that of the normal control, and compared with the adiponectin (APN), which is a positive control, adiponectin (containing adiponectin) (hSCFs) culture medium was slightly reduced in the presence of 10% of the dermal papilla culture medium, but 25% and 50% of the hSCFs were similar or superior.

VEGF and HGF were 1.5-3 times more expressed than normal control and similar to (15, 25%) and excellent (50%) adiponectin (APN) positive control group.

On the other hand, TGF-b1 (Transforming growth factor), which is activated during hair loss, is similar to or lower than that of normal control. Especially, as the concentration increases, Respectively.

Claims (6)

1. A composition for promoting hair growth or hair growth comprising, as an active ingredient, a trimethylcyclohexene compound represented by the following formula [Chemical Formula 1]

The method according to claim 1, Wherein the concentration of the active ingredient is 1 to 100 [mu] M based on the total volume of the composition. The method according to claim 1, A composition for promoting hair growth or hair growth, wherein the composition is a pharmaceutical composition. The method according to claim 1, Wherein the composition is a cosmetic composition. The method according to claim 1, Wherein the composition is an external preparation for skin. The method according to claim 1, Wherein the composition further comprises vitamin D.

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