Meloside-containing composition for relieving hair loss and improving hair growth

Abstract

The present invention relates to a composition for relieving alopecia and improving hair growth containing Meloside, and more particularly, to a composition for relieving alopecia and improving hair growth containing Meloside, which enables hair papilla cells to grow / proliferate and inhibits the expression / activity of androgen receptors, as an active ingredient.

Description

Technical Field

[0001] This invention relates to a composition containing Meloside for relieving hair loss and improving hair regrowth, and more specifically, to a composition containing Meloside as an active ingredient for relieving hair loss and improving hair regrowth. Background Technology

[0002] The hair growth cycle consists of three phases: the anagen (growth phase), the catagen (transitional phase), and the telogen (resting phase), which repeat in a certain cycle to maintain hair growth and loss. Specifically, it can be divided into the anagen phase of hair growth, the catagen phase in which growth stops and the hair bulb atrophies, and the telogen phase in which the hair papilla begins to activate or produce hair, thereby causing existing hair to fall out.

[0003] The growth phase of hair is about 3-5 years for men and about 4-6 years for women. The regression phase lasts about 60-90 days, and the resting phase lasts about 90-120 days, after which hair loss occurs naturally.

[0004] Hair loss can be categorized into male pattern baldness, female pattern baldness, telogen effluvium, alopecia areata, and stress-related hair loss. Its causes are diverse, including genetics, environment, psychological factors, and hormonal changes. Hair loss refers to the simultaneous and multiple occurrences of hair follicles ceasing to grow and falling out, with an increasing proportion of these follicles having stopped growing.

[0005] In particular, male pattern baldness is known to be caused by hormonal influences. The male hormone testosterone is converted to dihydrotestosterone (DHT) by 5α-reductase, which directly acts on hair follicles. This DHT induces the hair follicles to enter the regression phase, leading to hair loss. Excessive DHT increases can exacerbate hair thinning and hair loss, accelerating hair thinning and loss in the crown area, thus worsening hair loss.

[0006] Furthermore, DHT, generated from testosterone, directly acts on the dermal papilla cells of hair follicles, increasing the expression of androgen receptors. If the androgen receptor, acting as a nuclear translocase, binds to DHT and translocates to the nucleus, it increases the expression of hair loss inducing factors such as Dickkopf1, TNF-α, TGF-α, and IL-1, and induces inhibition of Wnt signaling factors, thereby causing hair loss. Therefore, to treat hair loss caused by these factors, methods are used to adjust the expression level of androgen receptors in dermal papilla cells.

[0007] Based on this technical background, the inventors focused on developing compositions for alleviating hair loss and improving hair regrowth, particularly compositions for preventing male pattern baldness or improving hair regrowth. They discovered that compositions containing Meloside as an active ingredient have the effect of promoting the growth / proliferation of dermal papilla cells and inhibiting the expression / activity of androgen receptors, thus completing this invention.

[0008] [Existing Technical Documents]

[0009] Korean Patent Publication No. 10-2011-0064989 (June 15, 2011) Summary of the Invention

[0010] Technical issues

[0011] The main objective of this invention is to provide a composition containing Meloside, its isomers, its hydrates, its solvates, or salts thereof as active ingredients for relieving hair loss and improving hair regrowth.

[0012] Another object of the present invention is to provide a pharmaceutical, health food and cosmetic prepared from the said composition.

[0013] Technical solution

[0014] To achieve the aforementioned objective, the present invention provides a composition for relieving hair loss and improving hair regrowth, comprising Meloside represented by the following chemical formula 1, preferably Meloside A represented by the following chemical formula 2 and / or Meloside L represented by the following chemical formula 3, its isomers, its hydrates, its solvates or salts thereof as active ingredients.

[0015]

Chemical Formula 1

[0016]

[0017]

Chemical Formula 2

[0018]

[0019]

Chemical Formula 3

[0020]

[0021] The present invention provides a composition for preventing male pattern baldness or improving hair regrowth, comprising Meloside represented by chemical formula 1, preferably Meloside A represented by chemical formula 2 and / or Meloside L represented by chemical formula 3, its isomers, its hydrates, its solvates or salts thereof as active ingredients.

[0022] In addition, the present invention provides a composition containing Meloside represented by chemical formula 1, preferably Meloside A represented by chemical formula 2 and / or Meloside L represented by chemical formula 3, its isomers, its hydrates, its solvates or salts thereof as active ingredients for promoting the growth / proliferation of papillary cells and inhibiting the expression / activity of androgen receptors.

[0023] In addition, the present invention provides a pharmaceutical, health food and cosmetic prepared from a composition containing Meloside represented by chemical formula 1, preferably Meloside A represented by chemical formula 2 and / or Meloside L represented by chemical formula 3, its isomers, its hydrates, its solvates or salts thereof as active ingredients.

[0024] In addition, the present invention provides the use of Meloside, represented by chemical formula 1, preferably Meloside A, represented by chemical formula 2, and / or Meloside L, its isomers, its hydrates, its solvates, or salts thereof, as an active ingredient in the preparation of a composition for relieving hair loss and improving hair growth.

[0025] Invention Effects

[0026] The composition containing Meloside according to the present invention for relieving hair loss and improving hair regrowth has excellent efficacy in promoting the growth / proliferation of dermal papilla cells and inhibiting the expression / activity of androgen receptors. Therefore, it can be used to prepare cosmetics, pharmaceuticals, etc. for preventing hair loss or improving hair regrowth. Attached Figure Description

[0027] Figure 1 This is a graph illustrating the effect of Meloside A / Meloside L on the growth / proliferation of dermal papilla cells according to an embodiment of the present invention.

[0028] Figure 2 This is a graph illustrating the effect of Meloside A / Meloside L on the growth / proliferation of external root sheath cells according to an embodiment of the present invention.

[0029] Figure 3 The graph shows the effect of Meloside A / Meloside L on improving male pattern baldness according to an embodiment of the present invention, based on Western blot results.

[0030] Figure 4 It is Figure 3 The Western blot results were quantified and illustrated in a graph.

[0031] Figure 5This is a graph showing the effect of Meloside A / MelosideL on inhibiting androgen receptor nuclear translocation according to an embodiment of the present invention, as observed by fluorescence microscopy.

[0032] Figure 6 It is Figure 5 The results of fluorescence microscopy observations were quantified and illustrated in a graph.

[0033] Figure 7 The graph illustrates the effect of Meloside A / Meloside L on inhibiting DKK1 expression according to an embodiment of the present invention.

[0034] Figure 8 The graph illustrates the effect of Meloside A / Meloside L on increasing VEGF according to an embodiment of the present invention.

[0035] Figure 9 The graph illustrates the effect of Meloside A / Meloside L on inhibiting apoptosis caused by dihydrotestosterone according to an embodiment of the present invention.

[0036] Figure 10 The graph illustrates the effect of Meloside A / Meloside L on inhibiting apoptosis caused by DKK1 according to an embodiment of the present invention.

[0037] Figure 11 The graph illustrates the effect of Meloside A / Meloside L on relieving hair loss inflammation according to an embodiment of the present invention.

[0038] Figure 12 The graph shows the effect of Meloside A / MelosideL on inhibiting reactive oxygen species according to an embodiment of the present invention, as observed by fluorescence microscopy.

[0039] Figure 13 It is Figure 12 The results of fluorescence microscopy observations were quantified and illustrated in a graph. Detailed Implementation

[0040] Preferred form for carrying out the invention

[0041] Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. Generally, the nomenclature used in this specification is well-known and commonly used in the art.

[0042] The present invention will now be described in more detail.

[0043] In this specification, the term "about" used to express length, area, volume, time (period), concentration, content, temperature, quantity, percentage (%), multiple, etc., means that there is a maximum permissible error of 10% within the corresponding numerical value or range.

[0044] On one hand, the present invention relates to a composition containing Meloside represented by the following chemical formula 1, preferably Meloside A represented by the following chemical formula 2 and / or Meloside L represented by the following chemical formula 3, more preferably Meloside A, its isomer, its hydrate, its solvate or its salt as an active ingredient, specifically a composition for relieving hair loss and improving hair growth.

[0045]

Chemical Formula 1

[0046]

[0047]

Chemical Formula 2

[0048]

[0049]

Chemical Formula 3

[0050]

[0051] In addition, the present invention provides the use of Meloside, represented by the chemical formula 1, preferably Meloside A, represented by the chemical formula 2, and / or Meloside L, its isomers, its hydrates, its solvates, or salts thereof, as an active ingredient in the preparation of a composition for relieving hair loss and improving hair regrowth.

[0052] Meloside A (isovitexin 2”-O-glucoside) is represented by chemical formula 2, and Meloside L (isoorientin 2”-O-glucoside) is represented by chemical formula 3 (see https: / / pubchem.ncbi.nlm.nih.gov / compound / Meloside-A; https: / / pubchem.ncbi.nlm.nih.gov / compound / Meloside-L).

[0053] In this specification, isomers refer not only to diastereomers, including optical isomers, but also to conformation isomers (i.e., isomers with more than one chemical bond that differ only in their angles), position isomers (especially tautomers), or geometric isomers (e.g., cis-trans isomers).

[0054] In addition, in this specification, "hydrate" refers to a compound bound with water, and includes inclusion compounds in which there is no chemical bond between water and the compound.

[0055] Additionally, in this specification, "solvent" refers to a compound formed between molecules or ions of a solute and molecules or ions of a solvent.

[0056] In this invention, the hair can be any hair or hair on the body of mammals, including humans, such as head hair, eyebrows, eyelashes, mustache, beard, chest hair, back hair, arm hair, leg hair, pubic hair, nose hair, or ear hair, but is not limited thereto, and is preferably head hair.

[0057] In this invention, the composition may be characterized by promoting the growth / proliferation of dermal papilla cells and reducing the expression of androgen receptors.

[0058] In this invention, the composition may be characterized by preventing apoptosis of hair papilla cells, relieving scalp inflammation, and reducing the generation of reactive oxygen species, which are hair loss inducing factors.

[0059] In this invention, the composition is characterized by having an effective ingredient content of 10 to 1000 ppm, preferably 10 to 100 ppm, and most preferably 20 to 30 ppm, based on the total weight of the composition. When the content of one or more of Meloside, preferably Meloside A and Meloside L, is less than 10 ppm based on the total weight of the composition, the effect of alleviating hair loss and improving hair regrowth is weak. When it is greater than 100 ppm, the increase in effect is not significant compared to the increase in content, and it may reduce the stability of the dosage form and the user experience.

[0060] The composition can be commercialized in various dosage forms (cosmetics, health functional foods, pharmaceuticals, etc.), and appropriate dosage forms can be formulated considering the functionality, cost, and other conditions of the product to be achieved.

[0061] The composition, as a cosmetic composition, can be, for example, a solution, suspension, emulsion, paste, gel, cream, lotion, powder, shampoo, conditioner, and spray, etc., and its dosage form is not particularly limited and can be appropriately selected according to the purpose.

[0062] The composition, as a health functional food composition, for example, as a health supplement, can be in dosage form such as powder, granules, tablets, capsules, beverages, etc., but is not limited thereto.

[0063] The active ingredient may be a prodrug of Meloside. A prodrug is a drug that has undergone chemical modification to adjust its physical and chemical properties. It is a drug that does not exhibit physiological activity itself, but after administration, it can be converted into the original drug in the body through chemical or enzymatic action and exert its therapeutic effect.

[0064] The composition, as a pharmaceutical composition, can be in dosage forms such as tablets, ointments, lotions, gels, creams, sprays, suspensions, emulsions, patches, etc., but is not limited thereto.

[0065] Forms for implementing the invention

[0066] The present invention will now be described in more detail through embodiments. These embodiments are merely illustrative of the invention, and those skilled in the art should understand that the scope of the invention should not be construed as limited by these embodiments.

[0067] [Reagents]

[0068] Meloside A was purchased from ChemFace, and Meloside L was isolated / purified and used in the experiments according to the methods described later. Unless otherwise stated, the reagents used in the cell culture and experimental examples 1 and 2 described later are products of Sigma-Aldrich.

[0069] Isolation and purification of Meloside L

[0070] Meloside L was isolated / purified from Dianthus chinensis and applied experimentally. The aerial parts of Dianthus chinensis were extracted by reflux with 70% ethanol, and the soft extract, concentrated using a rotary vacuum distillation unit, was fractionated using hexane, ethyl acetate, and n-butanol. The ethyl acetate fraction was then subjected to column chromatography to separate single compounds, using silica gel and ODS (octadecylsilane) as the stationary phase. The separation was achieved by NMR (…). 1 H, 13 C10 NMR analysis was used to confirm the structure of the isolated / purified substance.

[0071] Cell Culture

[0072] Human hair dermal papilla cells (HDPCs) used in the following experimental cases were cultured in DMEM (Dulbecco's Modified Eagle Medium) supplemented with 10% FBS (fetal bovine serum, WELGENE) and 1% antibiotic (penicillin-streptomycin, WELGENE) and cultured in an incubator at 37°C, 5% CO2, and under humid conditions.

[0073] The human hair follicle outer root sheath cells (ORS) used in the following experimental examples were cultured in MSCM supplemented with 5% FBS (fetal bovine serum, ScienCell), 1% MSCGS (mesenchymal stem cell growth supplement, ScienCell), and an antibiotic (penicillin-streptomycin, ScienCell). The cells were cultured on poly-L-Lysine-coated culture plates in an incubator at 37°C, 5% CO2, and under humid conditions.

[0074] Statistical Processing

[0075] Statistical analysis was performed using the t-test to examine the significance between the two groups and compared with the negative control group. A significant difference was considered to be found when *P<0.05, **P<0.01, or ***P<0.001.

[0076] [Experimental Example 1] Confirmation of the cytotoxicity of Meloside on dermal papilla cells and outer root sheath cells

[0077] To determine whether Meloside exhibits cytotoxicity in dermal papilla cells and outer root sheath cells, thereby affecting their growth / proliferation.

[0078] Specifically, after treating HDPCs (as dermal papilla cells) or ORS (as outer root sheath cells) with Meloside A or Meloside L at concentrations of 10, 30, 50, and 100 ppm for one day, CCK-8 (Cell Counting Kit-8, Dojindo, EU) reagent was added to the treated HDPCs according to the manufacturer's protocol, and the absorbance was measured at a wavelength of 450 nm using a microplate reader (BioTeK, USA). The growth / proliferation of dermal papilla cells treated with Meloside was confirmed by comparison with a negative control group (treated with DMEM instead of Meloside A or Meloside L). Figure 1 The results of HDPC processing are shown in the image. Figure 2 The result of processing ORS is shown in the figure.

[0079] like Figure 1 and Figure 2 As shown, HDPCs and ORS treated with Meloside A or Meloside L exhibited the same degree of cell growth / proliferation as the negative control group, thus confirming that there were no cytotoxic effects such as reducing the growth / proliferation of dermal papilla cells and outer root sheath cells.

[0080] [Experimental Example 2] Confirmation of the effect of Meloside treatment on improving male pattern baldness

[0081] In this study, the effects of Meloside (specifically, Meloside A and Meloside L) on androgen receptor expression were evaluated to confirm whether it has the effect of improving male pattern baldness.

[0082] Specifically, the following groups were prepared: (i) a negative control group, in which HDPCs (dermal papilla cells) were not treated with DHT (dihydrotestosterone) and Meloside A and Meloside L; (ii) an untreated group, in which HDPCs were treated with 1 μM DHT and Meloside A and Meloside L were not treated; (iii) a positive control group, in which HDPCs were treated with 1 μM DHT and 100 μM minoxidil was used instead of Meloside A and Meloside L; and (iv) a treated group, in which HDPCs were treated with 1 μM DHT to activate the androgen receptor, and then Meloside A or Meloside L was treated at a concentration of 30 or 100 ppm for one day. The HDPCs from (i) to (iv) were then dissolved, and the expression level of the androgen receptor (AR) was confirmed by Western blotting. To analyze the relative expression of each reagent, the expression level of actin was used as a baseline for correction. Figure 3 The image shows the expression confirmed by Western blotting. Figure 4 The results show the results of adjusting the relative expression of AR based on actin expression levels.

[0083] like Figure 3 and Figure 4 As shown, the decrease in androgen receptor (AR) expression in HDPC treated with Meloside A or Meloside L concentration-dependently reduces the formation of the AR-DHT complex, thereby improving hair loss (preventing hair loss, etc.) as AR activity decreases.

[0084] In addition, it was confirmed that Meloside A was significantly more effective than Meloside L in reducing androgen receptor (AR) expression.

[0085] [Experimental Example 3] Confirmation of the effect of Meloside treatment on inhibiting androgen receptor nuclear translocation

[0086] Androgen receptors, as transcription factors, are proteins that adhere to specific sites on DNA to regulate transcription. Specific transcription factors mediate the activation or repression of specific genes.

[0087] It is known that nuclear translocation of the androgen receptor inhibits genes in the Wnt signaling system that suppress hair loss and increases the expression of DKK1, a hair loss-inducing protein (Cell Death & Disease volume 11, Article number:407(2020)).

[0088] To confirm whether androgen receptors are involved in the transcription of hair loss-related genes, the recombinant plasmid EGFP-AR (Addgene) was used, and the degree of androgen receptor nuclear translocation in dermal papilla cells was measured using an intracellular injection method.

[0089] As the untreated group, DHT was treated alone. As the negative control group, neither DHT nor the reagents were treated. As the experimental group, DHT was treated with 100 ppm of Meloside A or Meloside L, and the results were measured using a fluorescence microscope at Ex 485 nm and Em 535 nm 24 hours later. Figure 5 The results observed using a fluorescence microscope are shown in the figure. Figure 6 The figure shows the number of cells with androgen receptor translocation in the treatment group compared to the untreated group.

[0090] like Figure 5 and Figure 6 As shown, DHT-induced androgen receptor translocation was confirmed to be inhibited by Meloside A and Meloside L. The androgen receptor translocation inhibition effect was confirmed to alleviate hair loss by reducing the expression of hair loss-inducing genes. The efficacy of Meloside A was confirmed to be superior to that of treatment with Meloside L.

[0091] [Experimental Example 4] Confirmation of the effect of Meloside treatment on inhibiting DKK1 expression

[0092] The study confirmed whether Meloside has the ability to regulate the expression of DKK1, which is the inferior stage of the androgen receptor.

[0093] After treating HDPCs with DHT to increase DKK1 expression, they were then treated with Meloside A or Meloside L at concentrations of 30 or 100 ppm for one day to obtain cell supernatants. For comparison, HDPCs treated with DHT (dihydrotestosterone) but not with Meloside A or Meloside L served as the untreated group, while HDPCs treated with 100 μM minoxidil served as the positive control group.

[0094] The expression level of DKK1 in the obtained cell supernatant was confirmed by enzyme-linked immunosorbent assay (ELISA). Figure 7 The results are shown in the figure.

[0095] like Figure 7 As shown, it was confirmed that in HDPC treated with Meloside A, the expression level of DKK1 decreased in a concentration-dependent manner, which reduced the inhibition of the Wnt signaling system and thus had a hair loss-relieving effect, but had no effect in Meloside L.

[0096] [Experimental Example 5] Confirmation of the effect of Meloside on VEGF

[0097] To confirm the effects of Meloside A and Meloside L on promoting hair growth, the expression level of VEGF (vascular endothelial growth factor) was evaluated. VEGF is a growth factor known to promote hair growth by participating in angiogenesis and vasodilation.

[0098] HDPC was treated with Meloside A or Meloside L at concentrations of 30 or 100 ppm for one day to obtain cell supernatants. For comparison, HDPC untreated with Meloside A and Meloside L was used as a negative control, and HDPC treated with 100 μM minoxidil was used as a positive control.

[0099] The expression level of VEGF in the obtained cell supernatant was confirmed by enzyme-linked immunosorbent assay (ELISA). Figure 8 The results are shown in the figure.

[0100] like Figure 8 As shown, increased VEGF expression was confirmed in HDPC treated with Meloside A and Meloside L, which helps improve blood circulation and thus alleviates hair loss. Furthermore, Meloside A showed a better effect on improving blood circulation than Meloside L.

[0101] [Experimental Example 6] Confirmation of the effect of Meloside treatment on inhibiting apoptosis caused by dihydrotestosterone (DHT).

[0102] The protective effects of Meloside A and Meloside L against DHT-induced apoptosis in dermal papilla cells (HDPCs) were confirmed.

[0103] Specifically, HDPCs (dermal papilla cells) were treated with Meloside A or Meloside L at 30 or 100 ppm for one day, followed by apoptosis induced by DHT (dihydrotestosterone). For comparison, HDPCs untreated with Meloside A and Meloside L and untreated with DHT served as a negative control group; HDPCs untreated with Meloside A and Meloside L but treated with DHT served as an untreated group; and HDPCs treated with 100 μM minoxidil or 5 mM NAC (N-acetylcysteine) followed by DHT served as a positive control group.

[0104] CCK-8 (Cellcounting kit-8, Dojindo, EU) reagent was added to HDPC according to the manufacturer's instructions, and absorbance was measured at 450 nm using a microplate reader (BioTeK, USA). Cell viability of Meloside-treated dermal papilla cells was confirmed by comparison with a negative control group (treated with DMEM instead of Meloside A or Meloside L). Figure 9 The result is shown in the figure.

[0105] like Figure 9 As shown, a cytoprotective effect against DHT was confirmed in HDPC treated with Meloside A. Conversely, no cytoprotective effect against DHT was confirmed in HDPC treated with Meloside L.

[0106] [Experimental Example 7] Confirmation of the effect of Meloside treatment on inhibiting apoptosis caused by DKK1

[0107] The protective effects of Meloside A and Meloside L against DKK1, which induces apoptosis in dermal papilla cells (HDPCs), were confirmed.

[0108] Specifically, HDPCs (dermal papilla cells) were treated with Meloside A or Meloside L at 30 or 100 ppm for one day, followed by apoptosis induced by DKK1 (dickkopf WNT signaling pathway inhibitor 1). For comparison, HDPCs untreated with Meloside A and Meloside L and without DHT served as a negative control group; HDPCs untreated with Meloside A and Meloside L but treated with DHT served as an untreated group; and HDPCs treated with 100 μM minoxidil or 5 mM NAC followed by DHT served as a positive control group.

[0109] CCK-8 (Cellcounting kit-8, Dojindo, EU) reagent was added to HDPC according to the manufacturer's instructions, and absorbance was measured at 450 nm using a microplate reader (BioTeK, USA). Cell viability of Meloside-treated dermal papilla cells was confirmed by comparison with a negative control group (treated with DMEM instead of Meloside A or Meloside L). Figure 10 The result is shown in the figure.

[0110] like Figure 10 As shown, a cell-protective effect against DKK1 was confirmed in HDPC treated with Meloside A. Conversely, no cell-protective effect against DKK1 was confirmed in HDPC treated with Meloside L.

[0111] [Experimental Example 8] Confirmation of the effect of Meloside treatment on alleviating hair loss inflammation

[0112] The study confirmed whether Meloside A and Meloside L have anti-inflammatory effects.

[0113] Specifically, the following groups were prepared: (i) negative control group, in which HDPCs (hair papilla cells) were not treated with DHT (dihydrotestosterone) and Meloside A and Meloside L; (ii) untreated group, in which HDPCs were treated with 100 μg / mL DHT and Meloside A and Meloside L were not treated; (iii) positive control group, in which HDPCs were treated with 100 μM minoxidil or 20 μM dexamethasone (DEX); and (iv) treatment group, in which HDPCs were treated with 100 μg / mL DHT to increase the expression of IL-6, and then Meloside A or Meloside L was treated at a concentration of 30 or 100 ppm for one day, respectively. The cell supernatants of HDPCs in (i) to (iv) were obtained, and the expression of IL-6 (interleukin 6) was confirmed by enzyme-linked immunosorbent assay (ELISA). In order to analyze the expression of each reagent, calibration was performed using the standard reagent of IL-6 as a benchmark. Figure 11 The result is shown in the figure.

[0114] like Figure 11 As shown, it was confirmed that the reduction in IL-6 expression in HDPC treated with Meloside A reduced DHT-induced inflammation, thereby alleviating scalp inflammation by decreasing IL-6 expression. Conversely, it was confirmed that HDPC treated with Meloside L did not reduce IL-6 expression.

[0115] [Experimental Example 9] Confirmation of the effect of Meloside treatment on the inhibition of dihydrotestosterone (DHT)-induced reactive oxygen species (ROS).

[0116] The inhibitory effect of HDPCs, used as human dermal papilla cells, on intracellular reactive oxygen species was evaluated. The assay was performed by utilizing the principle that the non-fluorescent substance 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA) reacts with intracellular free radicals and is oxidized to 2',7'-dichlorofluorescein (DCF), which is a fluorescent substance.

[0117] Human dermal papilla cells were divided into 5.5 x 10⁻⁶ cells. 4Cells were divided into 12-well plates and cultured at 37°C with 5% CO2 for 24 hours. Then, each well was treated with reagents prepared at different concentrations, and cultured at 37°C with 5% CO2 for 23 hours. The experiments included treatment groups (Meloside A or Meloside L treated with DHT at 30 or 100 ppm, respectively), control groups (treated with 100 μg / mL DHT but not Meloside A or Meloside L), and untreated groups (treated with neither Meloside A nor Meloside L and not DHT). Additionally, as positive controls, groups treated with 5 mM NAC or 10 μM minoxidil were used instead of Meloside A or Meloside L. After treating wells with 100 μg / mL DHT or Meloside and culturing for 24 hours, 2',7'-dichlorodihydrofluorescein diacetate (DCF-DA, a fluorescent probe) was added, and the reaction was carried out at room temperature for 30 minutes. After removing DCF-DA and washing twice with PBS, fluorescence microscopy was used to image cells at Ex 485 nm and Em 535 nm for 30 minutes at 10-minute intervals. The level of intracellular reactive oxygen species (ROS) generation was measured using ImageJ on a sample of 100 cells to confirm whether Meloside has the effect of reducing ROS. Figure 12 and Figure 13 The results are shown in the figure.

[0118] like Figure 12 and Figure 13 As shown, it was confirmed that in HDPC treated with Meloside A and Meloside L, the generation of reactive oxygen species (ROS) was reduced, thereby inhibiting the hair loss-inducing factor ROS induced by DHT, thus achieving a hair loss improvement effect (preventing hair loss, etc.).

[0119] Although specific portions of the invention have been described in detail above, it should be understood by those skilled in the art that these specific techniques are merely preferred embodiments and the scope of the invention is not limited thereto. Therefore, the essential scope of the invention will be defined by the appended claims and their equivalents.

Claims

1. A composition for relieving hair loss and improving hair regrowth, comprising Meloside, its isomer, its hydrate, its solvate, or a salt thereof, represented by the following chemical formula 1, as an active ingredient: [Chemical Formula 1] 2. The composition according to claim 1, characterized in that, The Meloside is selected from one or more of the group consisting of Meloside A represented by the following chemical formula 2 and Meloside L represented by the following chemical formula 3: [Chemical Formula 2] 【Chemical Formula 3】 3. The composition according to claim 1 or 2, characterized in that, The composition is used to prevent or improve male pattern baldness.

4. The composition according to claim 1 or 2, characterized in that, The composition promotes the growth of dermal papilla cells.

5. The composition according to claim 1 or 2, characterized in that, The composition reduces the expression of androgen receptors.

6. The composition according to claim 1 or 2, characterized in that, The composition prevents apoptosis of dermal papilla cells.

7. The composition according to claim 1 or 2, characterized in that, The composition relieves scalp inflammation.

8. The composition according to claim 1 or 2, characterized in that, The composition reduces the generation of reactive oxygen species, which are hair loss inducing factors.

9. The composition according to claim 1 or 2, characterized in that, The composition is a cosmetic composition, a health functional food composition, or a pharmaceutical composition.

10. Use of Meloside, represented by Formula 1, its isomers, its hydrates, its solvates, or salts thereof as an active ingredient in the preparation of a composition for alleviating hair loss and improving hair regrowth: [Chemical Formula 1] 11. The use according to claim 10, characterized in that, The Meloside is selected from one or more of the group consisting of Meloside A represented by the following chemical formula 2 and Meloside L represented by the following chemical formula 3: [Chemical Formula 2] 【Chemical Formula 3】

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