Novel compound obtained through biorenovation and composition comprising same for preventing or treating hair loss

Abstract

The present invention provides a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof, and a composition comprising same for preventing or treating hair loss.

Description

Novel compound obtained by biorenovation and composition for preventing or treating hair loss comprising the same

[0001] The present invention relates to a novel compound obtained by biorenovation and a composition for preventing or treating hair loss comprising the same.

[0002] Hair loss is a common concern. Androgenetic alopecia (AGA), characterized by scalp hair loss in both men and women as they age (i.e., male pattern baldness and female pattern baldness), is particularly common. Other hair loss concerns include alopecia areata (AA), a microscopic, inflammatory, usually reversible, localized form of hair loss, and alopecia associated with chemotherapy or radiation therapy (i.e., secondary alopecia).

[0003] Hair is typically divided into two types: terminal hair and vellus hair. Terminal hair is a thick, pigmented, long hair that grows from follicles deep within the dermis. Vellus hair is a fine, pigmentless hair that typically grows from smaller, more superficial follicles within the dermis. As hair loss progresses, terminal hair transforms into vellus hair, and the follicles correspondingly shrink.

[0004] Another factor in hair loss is changes in the hair growth cycle. Hair typically grows through three cycles: anagen (active hair growth), catagen (transitional phase), and telogen (a quiescent phase, in which the hair shaft falls out before new growth). Normally, about 88% of the hair on the scalp is in the anagen phase, only about 1% is in the catagen phase, and the remainder is in the telogen phase. As hair loss progresses, a sharply increasing proportion of hairs enter the telogen phase, and a corresponding decrease in the proportion of hairs in the active growth phase occurs.

[0005] Additionally, both the size and density of hair follicles are significantly reduced in association with alopecia. For example, balding individuals between the ages of 30 and 90 have been reported to have an average of only about 306 hair follicles per square centimeter. This is approximately 33% fewer than the average 460 hair follicles per square centimeter in non-balding individuals of the same age range. The combination of these factors contributes to baldness.

[0006] The most common method for addressing hair loss is medication. Various medications, from vitamins to hormones, have been tried, but their success rates are extremely limited.

[0007] Currently, patients with alopecia can attempt to regrow lost hair with repeated applications of topical steroids or maintain hair growth with topical minoxidil. While there have been some promising studies, there are no approved treatments capable of preventing or treating hair loss without the side effects of chemotherapy.

[0008] Therefore, there is still a need for safe and effective hair loss treatments without side effects.

[0009] [Prior Art Literature]

[0010] [Patent Document]

[0011] (Patent Document 1) Republic of Korea Publication No. 10-2013-0041848

[0012] The present invention has been devised to solve the above problems of the prior art.

[0013] The purpose is to provide a novel compound obtained through biorenovation that is also effective in preventing or treating hair loss.

[0014] In addition, the purpose is to provide a composition for preventing or treating hair loss that has excellent hair loss prevention or treatment effects and minimizes side effects.

[0015] The present invention

[0016] Provided is a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[0017] [Chemical Formula 1]

[0018]

[0019]

[0020] In addition, the present invention

[0021] A pharmaceutical composition for preventing or treating hair loss, comprising the compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, is provided.

[0022]

[0023] In addition, the present invention

[0024] A cosmetic composition for preventing or treating hair loss comprising the compound or a pharmaceutically acceptable salt thereof is provided.

[0025]

[0026] In addition, the present invention

[0027] A food composition for preventing or treating hair loss comprising the compound or a pharmaceutically acceptable salt thereof is provided.

[0028] The novel compound obtained through biorenovation of the present invention offers excellent effects in preventing or treating hair loss. Furthermore, its effectiveness in treating other diseases is also highly anticipated in the future.

[0029] The composition for preventing or treating hair loss of the present invention has an excellent effect of preventing or treating hair loss, and provides the effect of minimizing side effects.

[0030] Figure 1 shows the HPLC analysis results (HPLC analysis combined with flight electrospray mass spectrometry analyses) of Esculetin (ES) and its bioconversion-based derivative (ESBR (esculetin 6-O-phosphate)).

[0031] Figure 2 shows the composition of ESBR (esculetin 6-O-phosphate) in DMSO. 1 It shows the H-NMR spectrum,

[0032] Figure 3 shows the ESBR (esculetin 6-O-phosphate) in DMSO. 13 It shows the H-NMR spectrum,

[0033] Figure 4 shows the HMBC correlations of ESBR (esculetin 6-O-phosphate).

[0034] Figure 5 shows the ESBR (esculetin 6-O-phosphate) in DMSO. 31 It shows the P NMR spectrum,

[0035] Figure 6 is a graph showing the cell proliferation results of HFDP cells treated with various concentrations of ES, ESBR, and minoxidil.

[0036] The present invention relates to a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof:

[0037] [Chemical Formula 1]

[0038]

[0039] The compound represented by the above chemical formula 1 can be manufactured by a biorenovation bioconversion technique in which esculetin is added to Bacillus siamesis JD3-7 strain (KACC 92346P) and cultured.

[0040] The compound represented by the above chemical formula 1 or a pharmaceutically acceptable salt thereof has been confirmed to be excellent in preventing or treating hair loss. Furthermore, its effectiveness against other diseases is also expected to be significant in the future.

[0041] The pharmaceutically acceptable salt described above is not particularly limited and can be prepared and used without limitation in the form of a salt used in this field. In particular, the form of an organic or inorganic phosphate of the compound represented by Chemical Formula 1 can be usefully used. For example, the salt may include a pharmaceutically acceptable metal salt prepared using a base. The metal salt may be an alkali metal salt or an alkaline earth metal salt. The alkali metal salt or alkaline earth metal salt may be obtained, for example, by dissolving the compound of Chemical Formula 1 in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering out the undissolved compound salt, and evaporating and drying the filtrate. At this time, the metal salt may include lithium, sodium, potassium, calcium salt, etc.

[0042]

[0043] The composition may be a pharmaceutical composition, a cosmetic composition, or a food composition.

[0044] The above pharmaceutical composition, cosmetic composition, or food composition may contain 0.1 to 100 wt% of the compound represented by the above chemical formula 1, and may also contain various carriers and additives known in the art.

[0045] The above pharmaceutical composition can be prepared as a composition in a conventional dosage form by selecting one or more pharmaceutically acceptable conventional carriers or one or more additives from the compound represented by the above chemical formula 1.

[0046] The above carrier may be selected from one or more of diluents, lubricants, binders, disintegrants, sweeteners, stabilizers, and preservatives, and the additives may be selected from one or more of flavorings, vitamins, and antioxidants.

[0047] In the present invention, the carrier and additive are not limited in type, and for example, diluents include lactose monohydrate, trehalose, corn starch, soybean oil, microcrystalline cellulose, and D-mannitorl; lubricants include magnesium stearate and talc; and binders include polyvinylpyrrolidone (PVP), hydroxypropylcellulose (HPC), and the like.

[0048] In addition, disintegrants include carboxymethylcellulose calcium (Ca-CMC), sodium starchglycolate, polacrylin potassium, and cross-linked polyvinylpyrrolidone; sweeteners include white sugar, fructose, sorbitol, or aspartame; and stabilizers include carboxymethylcellulose sodium (Na-CMC), beta-cyclodextrin, etc. -cyclodextrin, white bee's wax, xanthan gum, etc.; preservatives include methyl p-hydroxy benzoate (methhlparaben), propyl p-hydroxybenzoate (propylparaben), potassium sorbate, etc.

[0049] The dosage of the above pharmaceutical composition may vary depending on the age, sex, and weight of the patient or animal to be treated, and above all, will depend on the condition of the subject to be treated, the specific category or type of the disease to be treated, the route of administration, and the properties of the therapeutic agent used. The above pharmaceutical composition may be appropriately selected depending on the absorption and excretion rate of the active ingredient in the body, the age and weight, sex, and condition of the patient or animal to be treated, the severity of the disease to be treated, etc., but may generally be administered at 0.001 to 1,000 mg / kg per day. The unit dosage form preparation formulated in this way may be administered multiple times at regular intervals as needed.

[0050] The above pharmaceutical composition may be administered individually as a preventive or therapeutic agent, or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents.

[0051] The above pharmaceutical compositions can be formulated and used as oral dosage forms such as powders, granules, tablets, capsules, troches, suspensions, emulsions, syrups, aerosols, etc., according to conventional methods. When formulating, they can be prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, troches, etc. Liquid preparations for oral administration include suspensions, oral solutions, emulsions, and syrups, and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives can be included.

[0052]

[0053] In one embodiment of the present invention, the composition may be a cosmetic composition. The cosmetic composition provides physiologically active effects such as hair loss prevention and hair growth.

[0054] The above cosmetic composition can be prepared in any formulation commonly manufactured in the art, and may be, for example, a solution, suspension, emulsion, paste, gel, water-soluble liquid, cream, essence, etc.

[0055] In addition, the cosmetic composition may contain adjuvants commonly used in the cosmetic field, such as hydrophilic or lipophilic active agents, preservatives, antioxidants, solvents, fillers, blocking agents, pigments, deodorants, dyes, etc. The amount of these various adjuvants is an amount commonly used in the field, and may be, for example, 0.01 to 30 wt% based on the total weight of the composition. However, in any case, the adjuvants and their proportions will be selected so as not to adversely affect the desirable properties of the cosmetic composition according to the present invention.

[0056] In the above cosmetic composition, when the formulation is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

[0057] In addition, when the formulation is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzoate, propylene glycol, 1,3-butylene glycol, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0058] In addition, when the formulation is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth may be used as a carrier component.

[0059] Additionally, when the formulation is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and especially in the case of a spray, a propellant such as chlorofluorohydrocarbon, protane / butane or dimethyl ether may be additionally included.

[0060]

[0061] In one embodiment of the present invention, the composition may be a food composition. The food composition may provide physiologically active effects such as hair loss prevention and hair growth. The food composition may be a health functional food.

[0062] The food composition may be tea, jelly, juice, extract, beverage, etc., and there is no limitation on the processed form as long as it contains the compound represented by the chemical formula 1. In addition, the food composition may be selected from the group consisting of general foods, such as ice cream, dairy products including milk, soy milk, and cheese, soy milk products, beverages, meat, sausage, bread, chocolate candies, snacks, confectionery, pizza, ramen, other noodles, various soups, beverages, tea, drinks, alcoholic beverages, and lactic acid bacteria preparations in capsule or stick packaging.

[0063] The above beverage composition may contain various flavoring agents, sweeteners, or natural carbohydrates as additional ingredients, as in conventional beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The sweetener may be a natural sweetener such as thaumatin or stevia extract, or a synthetic sweetener such as saccharin or aspartame.

[0064]

[0065] Hereinafter, the present invention will be described in more detail through examples. However, the following examples are intended to further illustrate the present invention, and the scope of the present invention is not limited by these examples. Those skilled in the art may appropriately modify or alter the following examples within the scope of the present invention.

[0066]

[0067] Example 1: Microbial culture and biorenovation of esculetin using the same.

[0068] The Bacillus siamesis JD3-7 strain (KACC 92346P) used in the bioconversion technique in the present invention was isolated from traditional soy sauce from Jeju Island. The strain was cultured in Luria-Bertani broth (BD Biosciences, San Jose, CA, USA) at 30°C and 200 rpm for 16 hours, and then centrifuged at 4,500 rpm for 15 minutes at 4°C to obtain a microbial precipitate excluding the culture medium. In addition, to remove the residual culture medium, the culture was washed twice with PG buffer (50 mM Phosphate buffer, 2% Glycerin). 100 mg of esculetin (Sigma-Aldrich, St. Louis, MO, USA) was added to the microorganisms suspended in the PG buffer, and the culture was cultured at 30°C and 200 rpm for 72 hours. The supernatant obtained by centrifugation was dried and used in the experiment.

[0069]

[0070] Experimental Example 1: Analysis of bioconverted esculetin derivatives using HPLC, LC / MS, and NMR analysis equipment.

[0071] (1) Experimental method

[0072] For HPLC analysis of esculetin (ES) and its bioconversion derivative (ESBR), a Shim-pack GIS C18 Column, 5 μm ODS, 250 X 4.6 mm ID (227-30106-08, Shimadzu Scientific Instruments, Inc., Baltimore, DC, USA) and a Shimadzu SpectroMonitor 3200 digital UV / Vis detector (228-42593-43, Shimadzu Scientific Instruments, Inc.) were used. Purified water (Solvent A) and acetonitrile (Solvent B, Sigma-Aldrich) with 0.1% trifluoroacetic acid (TFA, Samchun, Pyeongtaek, Korea) were used as mobile phase solvents. 10 μL of each sample diluted in purified water was injected at a column temperature of 40°C and the flow rate was set to 1.0 mL / min. Solvent A: Analysis was conducted for 30 minutes at a wavelength of 345 nm under gradient conditions from Solvent A: 90%, Solvent B: 10% to Solvent A: 0%, Solvent B: 100%.

[0073]

[0074] (2) Experimental results

[0075] When the product obtained by applying the biorenovation bioconversion technique to ES was analyzed using HPLC, two peaks were confirmed, and when the retention time was checked, it was confirmed that peak F1 detected at 5 minutes was a newly synthesized component by the bioconversion technique. In addition, it was confirmed that F2 detected at 6 minutes was ES, the substrate used in the bioconversion (Fig. 1 (A)). In addition, when the molecular weight of ESBR was confirmed using LC / MS, F2 had an m / z of 178.03, which was consistent with ES used as a substrate. In addition, F1 was confirmed to have an m / z of 257.99, which was confirmed to have the same molecular weight as the structure in which a phosphate group was bonded to the hydroxyl group of ES by the bioconversion technique (Fig. 1 (B)).

[0076] To analyze the exact structure of ESBR, 1D and 2D NMR were used. As a result, the chemical shifts of H-5 and carbon-5 (δH 7.45 ppm and δC-5 119.49 ppm) were downfield-shifted compared to the chemical shifts of esculetin. 13 In the C NMR spectrum, carbon-6 of ES shielded by phosphorylation was observed at 137.41 ppm (Figs. 2 and 3). In addition, 31 Based on the P NMR spectrum, phosphorylation of the hydroxyl group located at carbon 6 of ES was confirmed (Fig. 5). In addition, split carbon signals at carbons 5, 6, and 7 due to carbon-phosphorus bonds (JC-H 3.4-6.9 Hz) were also observed. 13 It was observed in the C NMR spectrum. Based on these results, the structure of ESBR was confirmed to be esculetin 6-O-phosphate.

[0077]

[0078] <Esculetin 6-O-phosphate(ESBR)>

[0079] 1H NMR (DMSO-d6, 400 MHz): δ 7.96 (1H, d, J= 9.5 Hz, H-4), 7.45 (1H, br d, H-5), 6.82 (1H, s, H-8), 6.23 (1H, d, J= 9.5 Hz, H-3) (Fig. 2A).

[0080] 13 C NMR (DMSO-d6, 100 MHz): δ 160.40 (Carbon-2), 153.29 (Carbon-7, d, J= 6.9 Hz), 151.20 (Carbon-8a), 144.27 (Carbon-4), 137.41 (Carbon-6, d, J= 6.7 Hz), 119.49 (Carbon-5, d, J = 3.4 Hz), 111.96 (Carbon-3), 110.45 (Carbon-4a), 103.91 (Carbon-8) (Fig. 2B, C).

[0081] 31 P NMR (DMSO-d6, 162 MHz): δ -4.26 (Fig. 2D).

[0082]

[0083] Experimental Example 2: Measurement of cell proliferation effect

[0084] (1) Cell culture

[0085] Human Follicle Dermal Papilla (HFDP) cells used in this experiment were obtained from PromoCell (Promocell, Heidelberg, DEU) and subcultured once every three days in a 37°C, 5% CO2 incubator using Dulbecco's Modified Eagle Medium (DMEM, Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum and 100 units / mL penicillin-streptomycin.

[0086]

[0087] (2) Measurement of cell proliferation effect

[0088] The proliferation rate of HFDP cells by sample treatment was confirmed through MTT assay. 2.0 х 10 cells were seeded in a 96-well plate. 4 After dispensing cells / well, they were cultured for 24 hours at 37°C and 5% CO2 conditions. After that, the untreated group, ES (esculetin), ESBR (esculetin derivative), and positive control minoxidil were treated at concentrations of 1, 10, 25, 50, and 100 μg / mL, and cultured for 48 hours. Then, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, Sigma-Aldrich) solution was added and reacted for 3 hours at 37°C and 5% CO2 conditions. After that, dimethyl sulfoxide (DMSO, Sigma-Aldrich) was added to the formed MTT formazan crystals to dissolve them, and transferred to a 96-well plate, and the absorbance was measured at 570 nm using an ELISA reader (Multiskan GO spectrophotometer, Thermo Fisher Scientific, Waltham, MA, USA).

[0089]

[0090] (3) Evaluation of cell proliferation rate

[0091] In order to measure the effect of ES derivatives on hair growth cell proliferation using the biorenovation bioconversion technique, cell viability and proliferation rates were measured using the MTT assay in HFDP cells treated with ESBR, ES, and the positive control minoxidil. As a result, the minoxidil-treated group showed cell proliferation rates of 18.1%, 25%, 10.8%, 6.4%, and -0.2%, and the cell proliferation rate tended to decrease in a concentration-dependent manner. The substrate ES used in the biorenovation bioconversion technique showed an increase rate of 0.9%, 7.7%, 8.5%, 16.5%, and 25.1%, and although the proliferation rate was not large at the measured concentrations, the cell proliferation tended to increase in a concentration-dependent manner. Finally, the cell proliferation rates of the novel compound ESBR were 36.8%, 34.3%, 26.3%, 21.5%, and 19.5%, showing the most excellent hair growth cell proliferation effect among the three samples at the same concentration (Fig. 6).

Claims

1. A pharmaceutical composition for preventing or treating hair loss, comprising a compound represented by the following chemical formula 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier: [Chemical Formula 1] 2. In paragraph 1, A pharmaceutical composition for preventing or treating hair loss, characterized in that the compound represented by the above chemical formula 1 is manufactured by a biorenovation technique in which esculetin is added to Bacillus siamesis JD3-7 strain (KACC 92346P) and cultured.

3. In paragraph 1, A pharmaceutical composition for preventing or treating hair loss, characterized in that the pharmaceutically acceptable salt is an organic phosphate or inorganic phosphate of a compound represented by chemical formula 1.

4. A cosmetic composition for preventing or improving hair loss, comprising the compound of paragraph 1 or a pharmaceutically acceptable salt thereof.

5. A food composition for preventing or improving hair loss, comprising the compound of paragraph 1 or a pharmaceutically acceptable salt thereof.

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