KR102076636B1 - Composition containing trimethyl cyclohexen compound for promoting hair sprouting or hair growth - Google Patents

Abstract

The present invention relates to a composition for promoting hair growth or hair growth comprising a trimethyl cyclohexene compound as an active ingredient, and the composition according to the present disclosure promotes hair induction cells and hair follicle activating factors, thereby promoting hair growth or hair growth. Has Therefore, the composition of the present disclosure may be variously used as a pharmaceutical composition or a cosmetic composition in a field such as hair loss treatment.

Description

Composition for promoting hair growth or hair growth comprising trimethyl cyclohexene compound as an active ingredient {COMPOSITION CONTAINING TRIMETHYL CYCLOHEXEN COMPOUND FOR PROMOTING HAIR SPROUTING OR HAIR GROWTH}

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

Human hair is primarily responsible for the defense function to protect the scalp from external stimulus such as ultraviolet rays and aesthetic function to express the external image of the individual. However, in modern society, hair loss is frequently caused by natural factors such as environmental pollution, strong ultraviolet rays, and physiological factors such as stress and hormonal imbalance. Therefore, the need for materials for the prevention and treatment thereof is urgently needed.

Hair follicles, the physiological organs in the body that make up the hair, form the hair during the postnatal development, the anagen phase in which the hair grows actively, the catagen phase in which the hair degenerates, and the hair fall out. The hair cycle is divided into a telogen phase, an exogen phase in which hair loss is maintained, and is involved in hair growth, maintenance, and dropout. In many studies, the activity of hair follicles is caused by dermal papilla cells, and in particular, the proliferation and differentiation of dermal papilla cells is primarily involved in the progress of the hair growth cycle and hair formation. In the growth phase where hair is actively growing, vigorous proliferation and differentiation of dermal papilla cells occurs actively, and hair growth is stopped, and in the degenerative, resting and hair loss phases when hair loss occurs, the cells are killed.

Therefore, hair growth and hair loss are closely related to proliferation and death of dermal papilla cells. Therefore, it is suggested that prolonging the growth period by inhibiting the growth of hair papilla cells or inhibiting cell death and shortening the degenerative phase, resting period, and hair loss will improve and treat hair loss. It is feed. In addition, cell division and migration in the vicinity of the dermal papilla are closely related to the growth of the hair. In the growing phase, hair is newly generated from the dermal papilla, and the cells are activated by various cytokines and hormones, resulting in the movement of the cells into the dermal papilla. Will affect growth.

In addition, growth factors such as HGF, IGF-1, and VEGF are most strongly expressed in the growth phase among hair follicle activating factors, and in the degenerative and resting phases, hair growth can be maintained by inducing proliferation of hair papilla cells and maintaining the growth phase. It is known to help.

Representative drugs minoxidil and finasteride, which are currently approved by the FDA, have side effects such as systemic hair growth and decreased sexual function, and the mechanism of action of minoxidil is not clear. Therefore, it is necessary to develop a biological material that promotes proliferation of dermal papilla cells in hair follicles and increases expression of growth factors while having less concern about side effects.

Korean Unexamined Patent Publication No. 10-2011-0000433

As discussed above, growth factors related to the growth of papillary cells and hair growth are very important factors in normal hair maintenance, hair loss inhibition and hair growth. In this regard, the present inventors have completed the present invention by experimentally confirming that trimethyl cyclohexene compound promotes proliferation of dermal papilla cells and increases expression of growth factors such as IGF-1.

Accordingly, it is an object of the present invention to provide a composition for promoting hair growth or hair growth comprising a trimethyl cyclohexene 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 trimethyl cyclohexene compound of the present invention as an active ingredient, promotes the growth of dermal papilla cells and the expression of growth factors, for the purpose of inhibiting hair loss and hair growth treatment It can be usefully used as a pharmaceutical composition, a cosmetic composition or an external preparation for skin.

1 is a composition containing a trimethyl cyclohexene compound (hereinafter referred to as PAC-6) according to an embodiment of the present invention, and a positive control adiponectin (APN) to the dermal papilla cells for proliferation and cytotoxicity analysis results. It is shown.
2 is a composition comprising a trimethyl cyclohexene compound (hereinafter referred to as PAC-6) according to an embodiment of the present invention, and the degree of hair growth activating factor expression of adiponectin (APN) as a positive control group (Hair growth-related gene expressions) ) Shows the result of the measurement.

Term Definition

In the present specification, when a part is said to "include" a certain component, it means that it can further include other components, without excluding the other components unless otherwise stated.

Illustrative Of embodiments  Explanation

Hereinafter, the present invention will be described in detail.

In exemplary embodiments of the present invention, the present invention is a composition for promoting hair growth or hair growth comprising a trimethyl cyclohexene compound represented by Formula 1 as an active ingredient.

[Formula 1]

The IUPAC name of the trimethyl cyclohexene compound of Formula 1 is 3- (2,6,6-trimethyl-cyclohex-1-enyl) -acrylic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester ( 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 trimethyl cyclohexene compound presented in the present invention contains an isomer. The “isomers” here are not only optical isomers (eg, essentially pure enantiomers, essentially pure diastereomers, or mixtures thereof), but also conformation isomers. ) (Ie, isomers that differ only in the angle of one or more chemical bonds), position isomers (especially tautomers) or geometric isomers (eg, cis-trans isomers).

The compound represented by Chemical Formula 1 is prepared by reacting a pyranone derivative compound of Chemical Formula 2 with a cyclohexenyl ester compound of Chemical Formula 3, as shown in Scheme 1 below:

Scheme 1

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

Referring to 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 cyclohexenyl ester compound.

At this time, in the pyranone compound of Formula 2, X is a halogen element, wherein the halogen element may be F, Cl, Br, or I, preferably Cl. The compound of formula (2) may be purchased and used commercially or can be prepared directly.

In the embodiment of the present invention, 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one in which X is Cl is used, which is 5-hydroxy-2- (hydroxymethyl) -4H-pyran. The 4-one was directly prepared by reacting with thionyl chloride (SOCl ₂ ).

In addition, the cyclohexenyl ester compound of formula 3 is 3- (2,6,6-trimethyl-cyclohex-2-enyl) -propenoic acid (3- (2,6,6-timethyl-cyclohex-2-enyl) as an ionized salt of propenoic acid, 3- (2,6,6-trimethyl-cyclohex-2-enyl) -propenoic acid (3- (2,6,6-timethyl-cyclohex-2-enyl) propenoic acid) It exists in the form which the cation (M +) couple | bonded with the carboxy group of.

The cation (M ⁺ ) is one kind selected from the group consisting of Li ⁺ , Na ⁺ , K ⁺ . Preferably, 3- (2,6,6- trimethyl-bicyclo hex-2-enyl) - pensan Pro (3- (2,6,6-timethyl-cyclohex -2-enyl) propenoic acid) and ions of Na ⁺ It may be a binder, to prepare it, for example 3- (2,6,6-trimethyl-cyclohex-2-enyl) -propenoic acid (3- (2,6,6-timethyl-cyclohex-2-enyl) It can be obtained by dissolving propenoic acid) and sodium hydroxide together in methanol to ionize and distilling methanol.

In a preferred embodiment of the invention, 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one as a compound of formula 2 and sodium 3- (2,6,6-trimethyl-cyclo as a compound of formula 3 A hydroxy pyranone compound of Formula 1 was prepared through ester linkage of hex-2-enyl) -propeneoate (Sodium 3- (2,6,6-timethyl-cyclohex-2-enyl) propenoate).

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

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

At this time, the solvent may be any solvent that can sufficiently dissolve the compounds of Formulas 2 and 3, for example, N, N-dimethylformamide (N, N-dimethylformamide, DMF), tetrahydrofuran (THF), dimethyl sulfoxide 1 type is selected from the group consisting of a side (DMSO), acetonitrile, dioxane, benzene, toluene, ether, methanol, hexane, cyclohexane, pyridine, N-methylpyrrolidone and combinations thereof, preferably DMF Use

After the reaction, the solvent is distilled and then subjected to a post-treatment process such as washing, drying, and purification to obtain a high purity compound.

When the trimethyl cyclohexene compound according to the present invention is included as an active ingredient, it has an effect of promoting the growth of dermal papilla cells and promotes the expression of growth factors IGF-1, VEGF, and HGF, thereby promoting hair production.

In the present specification, in particular, the effect of the composition in a test example described later, the human dermal papilla cells derived from humans, not animals such as rats, when applying the composition to the actual 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 from 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.

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

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

The pharmaceutical composition for promoting hair growth or hair growth according to an embodiment of the present invention is a preservative, stabilizer, hydrating or emulsifying accelerator, pharmaceutical supplements such as salts and / or buffers for controlling osmotic pressure and other therapeutically useful substances And may be formulated in various oral or parenteral dosage forms according to conventional methods.

The oral dosage forms include, for example, tablets, pills, hard and soft capsules, solutions, suspensions, emulsifiers, syrups, powders, powders, fine granules, granules, pellets, and the like, and these formulations include surfactants in addition to the active ingredients. , Diluents such as lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and glycine, and glidants such as silica, talc, stearic acid and its magnesium or calcium salts and polyethylene glycols. . Tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidine, optionally starch, agar, alginic acid or its sodium salt Pharmaceutical additives such as disintegrants, absorbents, colorants, flavors, and sweeteners. 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, a dosage form such as an injection, drop, ointment, lotion, gel, cream, spray, suspension, emulsion, suppository, or patch. May be, but is not limited thereto.

The pharmaceutical composition according to an embodiment of the present invention may be administered parenterally, rectally, topically, transdermally, subcutaneously, and the like. The pharmaceutical composition according to one embodiment of the invention may be administered topically to the scalp, for example.

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 depends on various factors such as less progression, onset, age, health condition, complications, etc. of the subject to be administered. Generally based on the composition may be 1 μg / kg to 100 mg / kg, such as 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 5 mg / kg to 10 mg / kg may be administered by dividing 1 to 3 times a day, the dosage is not intended to limit the scope of the invention in any way.

The hair growth or hair growth promoting composition according to an embodiment of the present invention may be a cosmetic composition, the appearance of the cosmetic composition contains a cosmetic or dermatologically acceptable medium or base. These are all formulations suitable for topical application, for example emulsions, suspensions, microemulsions, microcapsules, microgranules or ionics (liposomes) obtained by dispersing an oil phase in a solution, gel, solid, pasty anhydrous product, aqueous phase. And nonionic vesicle dispersants, or in the form of creams, skins, lotions, powders, ointments, sprays, or cone 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 the formulation, for example, supple cosmetics, astringent cosmetics, nourishing cosmetics, nutrition cream, massage cream, essence, eye cream, eye essence, cleansing It may be formulated into cosmetics such as creams, cleansing foams, cleansing water, packs, powders, body lotions, body creams, body oils and body essences.

When the formulation of the present invention is a paste, cream or gel, animal carriers, plant fibers, waxes, paraffins, starches, tracantes, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicas, talc or zinc oxide, etc. may be used as carrier components. 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, 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 or emulsifying agent is used as the carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 Fatty acid esters of, 3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan.

When the formulation of the present invention is a suspension, liquid carrier diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline Cellulose, aluminum metahydroxy, bentonite, agar or tracant and the like can be used.

When the formulation of the present invention is a surfactant-containing cleansing, the carrier component is an aliphatic alcohol sulfate, an aliphatic alcohol ether sulfate, a sulfosuccinic acid monoester, isethionate, an imidazolinium derivative, a methyltaurate, a sarcosinate, a fatty acid amide. Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, linolin 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 the 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, sphingolipids and seaweed extract.

In addition to the said functional additive, you may mix | blend the component contained in a general cosmetic composition with the cosmetic composition of this invention as needed. In addition to the other components included, oils and fats, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, fungicides, antioxidants, plant extracts, pH adjusters, alcohols, pigments, flavorings, blood circulation Accelerators, cooling agents, limiting agents, purified water, and the like.

Furthermore, 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 include any of cosmetics and medicines of various formulations as a generic term, which may include anything applied outside the skin. have.

In one embodiment, the composition may further comprise vitamin D. The composition may further supply vitamin D, thereby providing nutrients to the hair follicles. Their amount is preferably 0.0001 to 10% by weight, in particular 0.01 to 1% by weight of the total composition.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following practice is provided only for the purpose of illustration in order to help the understanding of the present invention, the scope and scope of the present invention is not limited thereto.

Production Example : 3- (2,6,6- Trimethyl - Cyclohex -One- Enil 5-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)

3- (2,6,6-trimethyl-cyclohex-1-enyl) -acrylic acid 5-hydroxy-4-oxo-4H-pyran-2 which is the trimethyl cyclohexene compound of formula 1 of the present invention according to Scheme 2 -Ylmethyl ester (3- (2,6,6-Trimethyl-cyclohex-1-enyl) -propenoic acid 5-hydroxy-4-oxo-4H-pyran-2-ylmethyl ester) was prepared.

Scheme 2

The detailed process of carrying out Scheme 2 is as follows.

50 g of 5-hydroxy-2- (hydroxymethyl) -4H-pyran-4-one (0.35 mmol) was dissolved in 250 ml of N, N-dimethylformamide, cooled in a 10 ° C. ice water bath, and 50 g of thionyl chloride ( 0.42 mol) was added dropwise for 30 minutes. After stirring for 2 hours at room temperature, the reaction solution was added to 2000 ml of ice water. The resulting solid was filtered and the 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. Drying in vacuo gave 39.5 g (70%) of 5-hydroxy-2- (chloromethyl) -4H-pyran-4-one of the formula 2-1 as a yellow solid.

Then 5 g (0.026 mol) of 3- (2,6,6-trimethyl-cyclohex-2-enyl) -propenoic acid (3- (2,6,6-timethyl-cyclohex-2-enyl) propenoic acid) 1.3 g (0.031 mol) of sodium hydroxide was dissolved in 40 ml of methanol, and after distilling methanol, the remaining residue was dissolved in 70 ml of N, N-dimethylformamide to prepare a compound of Chemical Formula 3-1.

To the 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 was added thereto and a 110 ° C oil bath. The mixture was heated and stirred for 2 hours at. The solvent was distilled off, the residue was dissolved in 300 ml of ethyl acetate, the ethyl acetate solution was washed with 5% hydrochloric acid and distilled water, and magnesium sulfate and activated carbon were added to dry and decolorize. The insolubles were then filtered and the filtrate was evaporated under reduced pressure to give 5.9 g (69% yield) of the reaction product as an off-white solid.

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

¹ H NMR (DMSO-d6, δ): 9.22 (s, 1H), 8.09 (s, 1H), 6.75 (m, 1H), 6.46 (s, 1H), 5.95 (d, 1H, J = 16.2 Hz) , 5.50 (s, 1H), 5.02 (s, 2H), 2.45 (m, 2H), 2.00 (m, 2H), 1.42 (m, 1H), 1.20 (m, 2H), 0.88 (s, 3H), 0.80 (s, 3H)

Example

Example  1: Proliferation and Cytotoxicity Analysis

Cell Culture and Differentiation

Human Subcutaneous Fat Cells (hSCFs) and Subcutaneous Fat Progenitor Cell Differentiation Medium Purchased from (NC, USA) and incubated in a CO ₂ incubator with 5% humidity. In order to induce differentiation, human subcutaneous fat (hSCFs) contains 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 1 μM troglitazone (Sigma-Aldrich, St. Louis, MO, USA) were incubated for 14 days in Dulbecco modified Eagles medium (DMEM; Lonza, Md, USA). Medium was changed every other day.

Human dermal papilla (HDP) and HDP maintenance medium were obtained from Cefobio Co. It was purchased from (Seoul, Korea) and cultured according to the manufacturer's instructions in a CO ₂ incubator with a humidity of 5%. To determine the effects of adiponectin (APN), subcutaneous fat culture medium containing the drug and human dermal papilla (HDP) medium were mixed at 1: 0.25, 1: 0.5, and 1: 1, respectively, followed by 2 for 6 days. Replace every day.

Cell proliferation and cytotoxicity assay

hSCFs viability 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, hSCFs were incubated for 7 days and treated with various concentrations of each chemical (adiponectin, hSCFs cultured with PAC-6) for 24 and 72 hours, respectively. 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 three times and the data shown as absorbance.

As a result of the experiment, it was found that the papillary cell proliferation effect was similar to that of the adiponectin, which is a positive control of the compound according to the present invention, and it was confirmed that the activity and hair production of keratin forming cells differentiated therefrom can be more effectively promoted.

In addition, it was confirmed that the cytotoxicity is also equivalent to that of the normal control group, which is not harmful to the human body.

Example  2: Analysis of Hair growth-related gene expressions

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

Total RNA was extracted from the dermal papilla 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, 1 μg of total RNA was used to synthesize complementary DNA (cDNAs) using PA, USA). Approximately 1 μg of cDNA sample and each TaqMan® probe (Life Technologies, Carlsbad, Calif., USA) are diluted in the reaction mixture of the Quantitect Probe PCR Kit (Qiagen, Valencia, Calif., USA), and PCR is run on a 7500 rapid real-time PCR system ( Fast real-time PCR system (Life Technologies, Carlsbad, Calif., USA). Each TaqMan® probe is as follows: glyceraldehyde-3-phosphate dehydrogenase (GAPDH; Part # 4352339E). All data are expressed in fold change relative to the GAPDH control and were obtained from three independent experiments.

Expression of growth factor IGF-1 (Insuline-like growth factor), VEGF (vascular endothelial growth factor), HGF (hepatocyte groeth factor) Referring to Figure 2, IGF-1 shows 2-3 times the expression compared to the normal control (control), subcutaneous fat (containing adiponectin) compared to the positive control adiponectin (APN) was applied (hSCFs) The culture medium content was 10% of the dermal papilla cell culture medium, but the effect was slightly decreased.

VEGF, and HGF also showed 1.5-3 fold expression compared to the normal control, similar (15, 25%) or superior (50%) to the positive control adiponectin (APN).

On the other hand, TGF-b1 (transforming growth factor), a factor that is activated during hair loss, was found to be similar to or lower than that of the normal control group. Especially, the higher the concentration, the lower the expression level. Indicated.

Claims (6)

To include a trimethyl cyclohexene compound represented by the formula (1) as an active ingredient,
A composition for promoting hair growth or hair growth wherein the concentration of the active ingredient is 1 to 100 μM based on the total volume of the composition:
[Formula 1]

delete The method of claim 1,
The composition for promoting hair growth or hair growth, characterized in that the pharmaceutical composition.
The method of claim 1,
The composition is a composition for promoting hair growth or hair growth, characterized in that the cosmetic composition.
The method of claim 1,
The composition for promoting hair growth or hair growth, characterized in that the skin external preparation.
The method of claim 1,
The composition for promoting hair growth or hair growth, characterized in that it further comprises vitamin D.

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