KR102119465B1 - Trimethoxy phenyl compound and composition containing the same for promoting hair sprouting or hair growth - Google Patents

Abstract

A composition for promoting hair growth or hair growth comprising a trimethoxy phenyl compound, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof and an active ingredient thereof, wherein the composition according to the present specification is a papillary cell and hair follicle By promoting the activation factor, it has the effect of promoting hair growth or hair growth accordingly. Therefore, the composition of the present specification may be variously used as a pharmaceutical composition or a cosmetic composition in fields such as hair loss treatment.

Description

Trimethoxy phenyl compound and composition for promoting hair growth or hair growth comprising the same {TRIMETHOXY PHENYL COMPOUND AND COMPOSITION CONTAINING THE SAME FOR PROMOTING HAIR SPROUTING OR HAIR GROWTH}

Disclosed herein is a trimethoxy phenyl compound, isomers thereof, pharmaceutically acceptable salts thereof, hydrates or solvates thereof, and compositions for promoting hair growth or hair growth comprising the same.

Human hair primarily serves as a protective function to protect the scalp from external stimuli such as ultraviolet rays and aesthetic functions that express the individual's external image. However, in modern society, hair loss occurs frequently due to environmental pollution, natural factors such as strong ultraviolet rays, and physiological factors such as stress and hormonal imbalance, and hair loss occurs regardless of age or gender, unlike the past limited to adult men. Accordingly, the need for materials for prevention and treatment thereof has been desperately needed.

The hair follicle, 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 the postnatal development process, a catagen phase in which the hair degenerates, and when the hair falls out It repeats the hair cycle, which is divided into a telogen phase and an exogen phase where hair loss occurs, and is involved in hair growth, maintenance, and dropping. By many studies, the activity of this hair follicle is caused by the papillary cells, and in particular, the proliferation and differentiation of the papillary cells is primarily involved in the progression of the hair growth cycle and hair formation. In the growth phase when hair is actively growing, the proliferation and differentiation of dermal papilla cells occurs actively, hair growth is stopped, and these cells are killed during the degenerative, resting, and hair loss phases when hair loss occurs.

Therefore, hair growth and alopecia are closely related to proliferation and death of hair papillae. Therefore, prolonging the growth phase or suppressing cell death and shortening the degeneration phase, the resting phase, and the alopecia to improve hair loss will be a treatment plan. Is fed. In addition, cell division and migration in the vicinity of the papilla are closely related to the growth of hair. In the growth phase, hair is newly generated from the papilla, and cells are activated by various cytokines, hormones, etc., and the movement of cells to the papilla appears. It will affect the growth of.

In addition, among the activation factors of hair follicles, growth factors such as HGF, IGF-1, and VEGF are most strongly expressed in the growth phase, and decrease in the degenerative and resting phases, thereby inducing proliferation of hair papilla cells and maintaining the growth phase, so that hair can be maintained normally. It is said to help.

Minoxidil and finasteride, which are currently approved by the FDA, have side effects such as systemic hair growth and decreased sexual function, and the specific mechanism of action of minoxidil has not been clearly identified. Therefore, there is a need to develop a biological material that promotes the proliferation of dermal papilla cells in the hair follicle and increases the expression of growth factors and has fewer side effects.

Korean Patent Publication No. 10-2011-0000433

As described above, growth factors related to proliferation of hair papilla cells and hair growth are very important factors for normal hair maintenance, hair loss inhibition, and hair growth. In response, the present inventors completed the present invention by preparing a novel trimethoxy phenyl compound and experimentally confirming that the compound promotes proliferation of dermal papilla cells and increases 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 comprising a novel trimethoxy phenyl compound, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof, and an active ingredient thereof in one aspect. Is to do.

In one aspect, the present invention provides novel trimethoxy phenyl compounds, isomers thereof, pharmaceutically acceptable salts thereof, hydrates or solvates thereof.

In one aspect, the present invention provides a method for producing a novel trimethoxy phenyl compound.

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

In one aspect, the novel trimethoxy phenyl compound of the present invention proliferates dermal papilla cells to a level similar to adiponectin secreted in the body, promotes expression of growth factors, and suppresses hair loss and hair growth treatment with these effects. It can be usefully used as a pharmaceutical composition for promoting hair growth or hair growth, as a cosmetic composition, or as an external preparation for skin.

1 is a composition comprising a trimethoxy phenyl compound (PAC-16742, hereinafter referred to as PAC-2) according to an embodiment of the present invention, and the positive control adiponectin (adiponectin, APN) treated on the papillary cells to proliferate and It shows the results of cytotoxicity analysis.
Figure 2 is a composition comprising a trimethoxy phenyl compound (PAC-16742, hereinafter referred to as PAC-2) according to an embodiment of the present invention, and the positive control adiponectin (adiponectin, APN) hair follicle activation factor expression level (Hair growth-related gene expressions).
Figure 3 shows the NMR results of the trimethoxy phenyl compound (PAC-16742, hereinafter referred to as PAC-2) according to an embodiment of the present invention (represented by Sample Name 742).

Term Definition

In the present specification, when a part “includes” a certain component, it means that the component may further include other components, rather than excluding other components, unless otherwise specified.

Exemplary Of implementations Explanation

Hereinafter, the present invention will be described in detail.

In exemplary embodiments of the present invention, the present invention is a trimethoxy phenyl compound represented by Formula 1 below, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or solvate thereof.

“Isomers” as used herein, in particular optical isomers (eg, essentially pure enantiomers, essentially pure diastereomers or mixtures thereof), as well as conformational isomers ( conformation isomers (i.e., isomers differing only in the angle of one or more chemical bonds), position isomers (especially tautomers) or geometric isomers (e.g., cis-trans isomers) do.

As used herein, “essentially pure,” when used in connection with, for example, enantiomers or diastereomers, contains at least about 90%, preferably at least about 95% of specific compounds that may exemplify enantiomers or diastereomers. , More preferably at least about 97% or at least about 98%, even more preferably at least about 99%, even more preferably at least about 99.5% (w/w).

"Pharmaceutically acceptable" herein is intended to be approved by the government or equivalent regulatory body for use in animals, more specifically in humans, by avoiding significant toxic effects when used in conventional medicinal dosages. It means that it can be received, approved, listed in the pharmacopeia or recognized as another general pharmacopeia.

“Pharmaceutically acceptable salt” as used herein refers to a salt according to an aspect of the present invention having pharmacologically acceptable and desirable pharmacological activity of a parent compound. The salt (1) is formed of an inorganic acid such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.; Or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl) Benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-Toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2,2,2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert Acid addition salts formed from organic acids such as butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; Or (2) a salt formed when the acidic proton present in the parent compound is substituted.

In the present specification, “hydrate” refers to a compound to which water is bound, and is a broad concept including an inclusion compound having no chemical bonding force between water and the compound.

As used herein, “solvate” refers to a higher order compound formed between molecules or ions of a solute and molecules or ions of a solvent.

In exemplary embodiments of the present invention, the present invention is a trimethoxy phenyl compound represented by Formula 1, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate or a solvate thereof as an active ingredient, or It is a composition for promoting hair growth.

[Formula 1]

The IUPAC name of the trimethoxy phenyl compound of Formula 1 is 3-(3,4,5-trimethoxy-phenyl)-acrylic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester (3- (3,4,5-Trimethoxy-phenyl)-acrylic 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 compound represented by Chemical Formula 1 is prepared by reacting a pyranone derivative compound of Chemical Formula 2 and a trimethoxy phenyl ester compound of Chemical Formula 3 as shown in Reaction Scheme 1 below:

[Scheme 1]

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

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

In this case, in the pyranone compound represented by Chemical Formula 2, X is a halogen element, and 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 product or by directly preparing it.

In one embodiment of the present invention, X may be 5-hydroxy-2-(chloromethyl)-4H-pyran-4-one where Cl is 5-hydroxy-2-(hydroxymethyl)-4H-pyran. It can be prepared by reacting -4-one with thionylchloride (SOCl ₂ ).

In addition, the trimethoxy phenyl ester compound of Formula 3 is an ionized salt form of 3-(3,4,5-trimethoxy-phenyl)-acrylic acid (3-(3,4,5-Trimethoxy-phenyl)-acrylic acid) As, as the carboxyl group of 3-(3,4,5-trimethoxy-phenyl)-acrylic acid (3-(3,4,5-Trimethoxy-phenyl)-acrylic acid), a cation (M+) may exist in a combined form. have.

The cation (M ⁺ ) is one selected from the group consisting of Li ⁺ , Na ⁺ , and K ⁺ . Preferably, it may be an ionic bond of 3-(3,4,5-trimethoxy-phenyl)-acrylic acid (3-(3,4,5-Trimethoxy-phenyl)-acrylic acid) and Na ⁺ , which is prepared In order to do this, for example, 3-(3,4,5-trimethoxy-phenyl)-acrylic acid (3-(3,4,5-Trimethoxy-phenyl)-acrylic acid) and sodium hydroxide are dissolved in methanol together and ionized. It can be obtained by distilling methanol.

In one preferred embodiment of the present invention, the compound of Formula 2 may be 5-hydroxy-2-(chloromethyl)-4H-pyran-4-one, and the compound of Formula 3 is sodium 3-(3,4,5- Trimethoxy-phenyl)-acrylic acid (Sodium 3-(3,4,5-Trimethoxy-phenyl)-acrylic acid). In addition, the trimethoxy phenyl compound of Formula 1 can be prepared through these ester bonds.

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

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

At this time, the solvent can be any one that can sufficiently dissolve the compounds of formulas 2 and 3, for example, N,N-dimethylformamide (N,N-dimethylformamide, DMF), tetrahydrofuran (THF), dimethyl sulfoxide Side (DMSO), acetonitrile, dioxane, benzene, toluene, ether, methanol, hexane, cyclohexane, pyridine, N-methylpyrrolidone, and combinations thereof. Can be used.

After the reaction, after distilling the solvent, a high-purity compound can be obtained through a general post-treatment process such as washing, drying, and purification.

The trimethoxy phenyl compound of Formula 1 according to the present invention can be applied to various fields, and can be preferably used as an active ingredient of a cosmetic composition. When the trimethoxy phenyl compound is included as an active ingredient, it has an effect of promoting the proliferation of dermal papilla cells, and promotes the expression of growth factors IGF-1, VEGF, and HGF, thereby promoting hair production.

In this specification, in particular, in the test examples described later, when the effect of the composition is tested on human dermal papilla cells, not animals such as rats, and the composition is applied to a real human body as a cosmetic or pharmaceutical composition, hair growth or hair growth It can be clearly seen that there is a promoting effect.

In one embodiment, the concentration of the active ingredient may be 1 to 100 μM based on the total volume of the composition, for example, 10 μM or more, 15 μM or more, 20 μM or more, 25 μM or more, 30 μM or more, 35 μM or more , Or 40 μM or more, and 90 μM or less, 80 μM or less, 70 μM or less, or 60 μM or less. In one embodiment of the present specification, preferably, the concentration of the active ingredient may be 50 μM.

When the concentration is less than 1 μM, the effect of promoting hair growth or hair growth may be negligible, and when it is greater 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 adds a preservative, a stabilizer, a hydrating agent or an emulsifying agent, a pharmaceutical auxiliary agent such as a salt and/or buffer for controlling osmotic pressure, and other therapeutically useful substances. It may contain, and may be formulated into various oral dosage forms or parenteral dosage forms according to conventional methods.

The oral dosage form, for example, tablets, pills, hard and soft capsules, liquids, suspensions, emulsifiers, syrups, powders, powders, granules, granules, pellets, etc., these formulations are surfactants in addition to the active ingredients , Diluents (e.g. lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine), lubricants (e.g. silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycols) . Tablets may also contain a binder such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, and optionally starch, agar, alginic acid or its sodium salt It may contain pharmaceutical additives such as disintegrants, absorbents, colorants, flavoring agents, and sweetening agents. The tablets can be prepared by conventional mixing, granulating or coating methods. In addition, the parenteral dosage form may be a transdermal dosage form, for example, injection, drip, ointment, lotion, gel, cream, spray, suspension, emulsion, suppository, patch, etc. However, it is not limited thereto.

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

The pharmaceutical composition may increase the expression of one or more of the growth factors IGF-1, VEGF and HGF, and the pharmaceutical composition may decrease the expression of TGF-b1, a factor that is activated when hair loss progresses.

Determination of the dosage of the active ingredient is within the level of those skilled in the art, and the daily dosage of the drug varies depending on various factors such as the degree of progression, onset time, age, health condition, and complications of the subject to be administered. In general, the composition may be 1 μg/kg to 100 mg/kg of the composition, 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/kg, preferably Can be administered by dividing 5mg/kg to 10mg/kg 1 to 3 times a day, the dosage is not to limit the scope of the present invention in any way.

The composition for promoting hair growth or hair growth according to an embodiment of the present invention may be a cosmetic composition, and the appearance of the cosmetic composition contains a cosmetic or dermatologically acceptable medium or base. These are all formulations suitable for topical application, e.g. solutions, gels, solids, dough anhydrous products, emulsions, suspensions, microemulsions, microcapsules, microgranules or ionic forms (liposomes) obtained by dispersing the oil phase in an aqueous phase. And non-ionic vesicle dispersants, or in the form of creams, skins, lotions, powders, ointments, sprays or conceal sticks. These compositions can be prepared according to conventional methods in the art. The composition according to the invention can 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, for example, softening makeup, converging makeup, nutrient makeup, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing It can be formulated into cosmetics such as 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 fibers, plant fibers, wax, paraffin, starch, tracant, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component. Can be.

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, and in particular, in the case of a spray, additionally chlorofluorohydrocarbon, propane /Propellant such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid ester.

When the formulation of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol as carrier components, suspensions such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxide, bentonite, agar or trakant, etc. can be used.

When the formulation of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivatives, methyltaurate, sarcosinate, fatty acid amide as a carrier component Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linoline derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

The cosmetic composition according to an embodiment of the present invention may further include a functional additive and a component included in a general cosmetic composition in addition to the active ingredient. The functional additive may include a component selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, polymer peptides, polymer polysaccharides, sphingo lipids, and seaweed extract.

The cosmetic composition of the present invention may also contain, in addition to the functional additives, components contained in a general cosmetic composition as necessary. Other ingredients included include oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, UV absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavors, blood circulation Accelerators, cooling agents, limiting agents, purified water, and the like.

Moreover, 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 is a generic term that may include anything applied from outside of the skin, and various types of cosmetics and pharmaceuticals may be included therein. have.

In addition, the composition for promoting hair growth or hair growth according to an embodiment of the present invention may be a food composition, and the food composition may be a liquid or solid formulation, tablets, capsules, soft capsules, pills, granules , Drink (drinks), diet bar, chocolate, caramel formulation or confectionery formulation, the formulation is not particularly limited. The food composition of the present invention may contain excipients, sugars, flavors, pigments, fats and oils, proteins, and the like in addition to the active ingredients.

In one embodiment, the composition may further include vitamin D. The composition can further supply nutrients to the hair follicles by further including vitamin D. The amount of these is preferably 0.0001 to 10% by weight, particularly 0.01 to 1% by weight of the total composition.

Hereinafter, the present invention will be described in more detail through the following examples. However, the following implementations are provided for the purpose of illustration only to aid understanding of the present invention, and the scope and scope of the present invention are not limited thereto.

Example

Example 1: 3 -(3,4,5- Trimethoxy -Phenyl)-acrylic acid 5-hydroxy-4-oxo-4 H -Pyran-2-ylmethyl ester (3-(3,4,5- Trimethoxy -phenyl)-acrylic acid 5- hydroxy -4- oxo -4H-pyran-2-ylmethyl ester)

3-(3,4,5-trimethoxy-phenyl)-acrylic acid 5-hydroxy-4-oxo-4H-pyran-2-yl, a trimethoxy phenyl compound of Formula 1 of the present invention according to Scheme 2 below Methyl ester (3-(3,4,5-Trimethoxy-phenyl)-acrylic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester) was prepared.

[Scheme 2]

The detailed process of performing Scheme 2 is as follows.

50g of 5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one (0.35mmol) was dissolved in 250ml of N,N-dimethylformamide, cooled in an ice-water bath at 10°C, and 50g of thionyl chloride ( 0.42 mol) was added dropwise over 30 minutes. After stirring at room temperature for 2 hours, a reaction solution was added to 2000 ml of ice water. The resulting solid was filtered, and solid (filtrate) was dissolved in 1000 ml of ethyl acetate. Magnesium sulfate and activated carbon were added, dried, decolorized, filtered, and the filtrate was concentrated and nucleic acid was added to obtain crystals. The product was dried under vacuum to obtain 39.5 g (70%) of 5-hydroxy-2-(chloromethyl)-4H-pyran-4-one of Chemical Formula 2-1 as a yellow solid.

Subsequently, 3-(3,4,5-trimethoxy-phenyl)-acrylic acid (3-(3,4,5-Trimethoxy-phenyl)-acrylic acid) (3-(2,6,6-timethyl-cyclohex -2-enyl) propenoic acid) 5 g (0.026 mol) and sodium hydroxide 1.3 g (0.031 mol) were dissolved in 40 ml of methanol, and after distilling methanol, the remaining residue was dissolved in 70 ml of N,N-dimethylformamide to obtain the formula 3-1. The compound was prepared.

To the prepared compound of Chemical Formula 3-1, 4.2 g (0.026 mol) of 5-hydroxy-2-(chloromethyl)-4H-pyran-4-one of Chemical Formula 2-1 prepared above was added, and an oil bath at 110° C. was added. And stirred for 2 hours. After distilling the solvent and dissolving the residue in 300 ml of ethyl acetate, the ethyl acetate solution was washed with 5% hydrochloric acid and distilled water, dried and decolorized by adding magnesium sulfate and activated carbon. Then, the insoluble matter was filtered and the filtrate was evaporated under reduced pressure to obtain 5.9 g (69% yield) of the reaction product as an off-white solid.

The NMR results of the off-white solid are shown in FIG. 3.

Test example

Test example 1: Proliferation and cytotoxicity analysis

Cell culture and differentiation

Human Subcutaneous Fat Cells (hSCFs) and subcutaneous fat progenitor cell differentiation media were used for ZenBio Inc. (NC, USA) and incubated in a CO ₂ incubator with a humidity of 5%. To induce differentiation, hSCFs are 10% fetal bovine serum (FBS, PAA, Pasching, Austria), 10 μg/ml insulin (Sigma-Aldrich, MO, USA), 0.5 mM 3-isobutyl-1-methylxanthine (IBMX; Sigma-Aldrich, St. Louis, MO, USA) 1 μM dexamethasone (DEX, Sigma-Aldric, MO, USA), St. Louis, MO, USA) and Dulbecco's modified Eagle's medium (DMEM; Lonza, Md, USA) containing 1 μM troglitazone (Sigma-Aldrich, St. Louis, MO, USA) for 14 days. Medium was changed every other day.

Human dermal papilla cells (hereinafter referred to as HDPs) and HDPs maintenance medium are Cefobio Co. (Seoul, Korea) and incubated according to the manufacturer's instructions in a CO ₂ incubator with a humidity of 5%.

Cell proliferation and cytotoxicity analysis

In order to know the degree of cell proliferation and cytotoxicity caused by PAC-2, HDPs were analyzed by treating hSCFs culture medium treated with adiponectin (hereinafter referred to as APN) and drug (PAC-2) as HDPs.

HDPs viability was measured according to the manufacturer's instructions using the EZ-Cytox cell viability assay kit (MTT assay, Daeil lab Service, South Korea). In summary, HDPs 24 hours after incubation were treated with APN 1, 5, 10 ng/ml as a positive control, and in the experimental group, the content of the hSCFs culture medium treated with PAC-2 (50 μM) was HDPs culture medium volume. It was treated for 10, 25, 50, and 100%, respectively, and treated for 24 hours and 72 hours. Then, EZ-Cytox solution (10 μl) was added to each well and incubated at 37° C. for 2 hours. Absorbance at 450 nm was measured using a spectrophotometer (Synergy H2, BioTek., VT, USA). All experiments were performed 3 times and the data are expressed as absorbance.

As a result of the experiment, as in the positive control APN, the hSCFs culture medium treated with the compound according to the present invention, PAC-2, showed no significance for proliferation or toxicity related to cell proliferation of HDPs. Therefore, it was confirmed that the compound according to the present invention exhibits hair growth or hair growth effect by promoting the activity of hair follicle activation factors in cells described later, not cell proliferation.

In addition, it was confirmed that the cytotoxicity was not harmful to the human body because it was considered to be at the same level as the normal control group.

Test example 2: Analysis of hair growth-related gene expressions

Real-time quantitative PCR (Real-time quantitative PCR (RT- qPCR ))

Total RNA was extracted from the papillary cells cultured in Example 1 above using TRIzol reagent (Life Technologies, Carlsbad, CA, USA), and the RevertAid first strand cDNA synthesis kit (Thermo Scientific, Pittsburgh, PA, USA) to synthesize complementary DNA (cDNAs), 1 μg of total RNA was used. About 1 μg of cDNA sample and each TaqMan® probe (Life Technologies, Carlsbad, CA, USA) was diluted in the reaction mixture of the Quantitect Probe PCR kit (Qiagen, Valencia, CA, USA), and PCR was performed on a 7500 fast real-time PCR system ( Fast real-time PCR system) (Life Technologies, Carlsbad, CA, USA). Each TaqMan® probe is as follows: glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Part # 4352339E). All data are expressed as fold change compared to the GAPDH control and were obtained in three independent experiments.

Growth factor IGF-1 (Insulin-like growth factor), VEGF (Vascular endothelial growth factor), HGF (Hepatocyte groeth factor) Referring to FIG. 2, in the case of IGF-1, the expression of 1.5-3 times as compared to that of the normal control (control), and compared with the application of the positive control APN at 1, 10 ng/ml, PAC-2 treatment When the hSCFs culture medium content was 10% based on the volume (ml) of the HDPs culture medium, the effect was somewhat lowered, but it was found to be similar or superior to the positive control group at 25 and 50%.

In the case of VEGF and HGF, the expression was 1.5-3 times higher than that of the normal control group, and it was similar (15, 25%) or superior (50%) to the positive control APN.

On the other hand, in the case of TGF-b1 (transforming growth factor), which is an active factor during hair loss progression, it was found to be similar to or lower than the normal control, especially when the concentration is increased from 10% to 25%. Showed a significant result of lowering.

Claims (11)

Trimethoxy phenyl compound represented by the following formula (1), its optical isomer, conformational isomer, geometric isomer, its pharmaceutically acceptable salt, hydrate or solvate thereof:
[Formula 1]

It is represented by the following scheme 1,
A method for preparing a trimethoxy phenyl compound of Formula 1 prepared by reacting a pyranone compound of Formula 2 and a trimethoxy phenyl ester compound of Formula 3 below:
[Scheme 1]

In Scheme 1, X is a halogen element and M is Li, Na or K.
A composition for promoting hair growth or hair growth comprising the trimethoxy phenyl compound represented by the following Chemical Formula 1, its optical isomer, conformational isomer, geometric isomer, pharmaceutically acceptable salt thereof, hydrate or solvate thereof as an active ingredient:
[Formula 1]

According to claim 3,
The composition for promoting hair growth or hair growth, characterized in that the concentration of the active ingredient is 1 to 100 μM based on the total volume of the composition.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, characterized by increasing the expression of one or more factors of IGF-1, VEGF and HGF.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, characterized by reducing the expression of TGF-b1.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, characterized in that the pharmaceutical composition.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, characterized in that the cosmetic composition.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, characterized in that the external preparation for skin.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, characterized in that the food composition.
According to claim 3,
The composition is a composition for promoting hair growth or hair growth, further comprising vitamin D.

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