Composition for repairing cell DNA damage comprising kaempferol glycoside compound

Abstract

The present application provides a composition for repairing cell DNA damage, which contains, as an active ingredient, a kaempferol compound extracted from Camellia oil meal discarded after extracting oil from Camellia seeds, and thus is friendly to the natural environment, safe to the human body due to few side effects, and capable of protecting DNA or cells damaged by photostimulation.

Description

Technical Field

[0001] The present application relates to a composition containing kaempferol glycoside compounds for repairing cell DNA damage, which is safe for the human body and can restore damaged DNA or skin cells caused by light stimuli such as ultraviolet rays. Background Art

[0002] Aging occurs due to various reasons. Among them, ultraviolet rays are considered the most common cause, and therefore, there are also many reports on skin photoaging caused by ultraviolet rays. When photo damage caused by ultraviolet rays occurs, DNA in the cell nucleus will be damaged, which will lead to cell damage with various results (cell death, cell aging, metabolic problems, etc.).

[0003] Specifically, ultraviolet rays are divided into UVA, UVB, and UVC according to wavelength. Among them, UVA has a long wavelength of 320 - 400 nm. When exposed to the skin for a long time, it can reach the dermis of the skin and cause skin irritation. UVB has a medium wavelength of 290 - 320 nm and can reach the basal layer of the epidermis or the upper layer of the dermis of the skin, and can cause skin irritation in a short time.

[0004] Therefore, a substance is needed to help restore damaged DNA, cells, especially skin cells caused by these UVA and UVB ultraviolet ray stimuli. Summary of the Invention

[0005] Technical Problem

[0006] In one aspect, the present application aims to provide a composition containing kaempferol glycoside compounds as active ingredients, which is safe for the human body with few side effects and can repair cell DNA damage.

[0007] In another aspect, the present application aims to provide a composition for repairing cell DNA damage, which contains kaempferol glycoside compounds extracted from Camellia japonica seed cake as active ingredients, and utilizes the waste resources discarded after extracting camellia seed oil.

[0008] Technical Solution

[0009] The present application provides a composition for repairing cell DNA damage, the composition containing kaempferol glycoside compounds represented by any one of the following Chemical Formulas 1 to 8, their isomers, their pharmaceutically acceptable salts, their hydrates or their solvates as active ingredients.

[0010]

Chemical Formula 1

[0011]

[0012] Chemical Formula 2

[0013]

[0014] Chemical Formula 3

[0015]

[0016] Chemical Formula 4

[0017]

[0018] Chemical Formula 5

[0019]

[0020] Chemical Formula 6

[0021]

[0022] Chemical Formula 7

[0023]

[0024] Chemical Formula 8

[0025]

[0026] Beneficial Effects

[0027] In one aspect, the composition for repairing cellular DNA damage disclosed in the present application contains kaempferol triglycoside compound or kaempferol tetraglycoside compound as an active ingredient, so that the damaged DNA or skin cells caused by light stimulation can be restored. Description of the Drawings

[0028] Figure 1 Shows molecular markers sensitive to DNA damage and repair according to an embodiment of the present application.

[0029] Figure 2 Shows γ-H2AX expression evaluation imaging according to an embodiment of the present application to confirm the efficacy of inhibiting DNA damage.

[0030] Figure 3 Shows the quantitative analysis results of γ-H2AX expression intensity according to an embodiment of the present application to confirm the efficacy of inhibiting DNA damage.

[0031] Figure 4 Shows the cell viability imaging under ultraviolet light according to an embodiment of the present application.

[0032] Figure 5Shows the results of quantitative analysis of cell viability under ultraviolet light according to an embodiment of the present application to analyze the cell count.

[0033] Figure 6 Shows the NMR information of the compounds represented by Chemical Formulas 1 to 4 according to an embodiment of the present application.

[0034] Figure 7 Shows the NMR information of the compounds represented by Chemical Formulas 5 to 8 according to an embodiment of the present application. Detailed Description of the Invention

[0035] In the present application, when a certain part "comprises" a certain component, unless otherwise specifically stated, it means that the part may further contain other components, rather than excluding other components.

[0036] Hereinafter, the present application will be described in detail.

[0037] In an exemplary embodiment of the present application, a composition for repairing cell DNA damage is provided, the composition comprising a kaempferol glycoside compound represented by any one of the following Chemical Formulas 1 to 8, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

[0038]

Chemical Formula 1

[0039]

[0040]

Chemical Formula 2

[0041]

[0042]

Chemical Formula 3

[0043]

[0044]

Chemical Formula 4

[0045]

[0046]

Chemical Formula 5

[0047]

[0048]

Chemical Formula 6

[0049]

[0050]

Chemical Formula 7

[0051]

[0052]

Chemical Formula 8

[0053]

[0054] According to the present application, by repairing the DNA of skin cells, not only can the damaged skin cells be restored, but also the skin cells that are about to be damaged can be prevented from being damaged in advance through DNA repair.

[0055] In another exemplary embodiment of the present application, a method for repairing DNA damage of cells is provided, which includes: administering or applying an effective amount of the above composition to a subject in need.

[0056] In another exemplary embodiment of the present application, the use of kaempferol glycoside compounds represented by any one of the following chemical formulas 1 to 8, their isomers, their pharmaceutically acceptable salts, their hydrates or their solvates in the preparation of the above composition for repairing DNA damage of cells is provided.

[0057] In another exemplary embodiment of the present application, the non-therapeutic use of kaempferol glycoside compounds represented by any one of the following chemical formulas 1 to 8, their isomers, their pharmaceutically acceptable salts, their hydrates or their solvates in repairing DNA damage of cells is provided.

[0058]

Chemical formula 1

[0059]

[0060]

Chemical formula 2

[0061]

[0062]

Chemical formula 3

[0063]

[0064]

Chemical formula 4

[0065]

[0066]

Chemical formula 5

[0067]

[0068]

Chemical formula 6

[0069]

[0070]

Chemical formula 7

[0071]

[0072]

Chemical formula 8

[0073]

[0074] In another exemplary embodiment of the present application, there is provided a kaempferol glycoside compound, an isomer thereof, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof for improving, preventing, or treating a disease caused by light stimulation, and the kaempferol glycoside compound is represented by any one of the following Chemical Formulas 1 to 8.

[0075]

Chemical Formula 1

[0076]

[0077]

Chemical Formula 2

[0078]

[0079]

Chemical Formula 3

[0080]

[0081]

Chemical Formula 4

[0082]

[0083]

Chemical Formula 5

[0084]

[0085]

Chemical Formula 6

[0086]

[0087]

Chemical Formula 7

[0088]

[0089]

Chemical Formula 8

[0090]

[0091] In the present application, the term "isomer" includes not only optical isomers, for example, essentially pure enantiomers, essentially pure diastereomers, or mixtures thereof, but also conformational isomers (i.e., isomers that differ only in the angle of one or more chemical bonds), positional isomers (especially tautomers), or geometric isomers (e.g., cis-trans isomers).

[0092] In the present application, "essentially pure" means, for example, when using the relevant enantiomers or diastereomers, the specific compound that can be exemplified by the enantiomers or diastereomers accounts for more than about 90% (w / w), preferably more than about 95% (w / w), more preferably more than about 97% (w / w), or more than 98% (w / w), still more preferably more than about 99% (w / w), and further preferably more than about 99.5% (w / w).

[0093] In the present application, "pharmaceutically acceptable" means that when used at conventional medicinal dosages, it avoids significant toxic effects, and thus can be used in animals (more specifically in humans), and it has obtained or can obtain approval from a government or equivalent regulatory agency, or is listed in a pharmacopoeia or known through other general pharmacopoeias.

[0094] In the present application, "medically acceptable salt" means a salt that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound according to one aspect of the present application. The salts may include: (1) acid addition salts formed from inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc., or acid addition salts formed from organic acids such as 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-ethanedisulfonic 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-butylacetic acid, dodecylsulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, mucic acid, etc.; or, (2) salts formed by substituting the acidic protons present in the parent compound.

[0095] In the present application, a hydrate refers to a compound combined with water, which is used in a broad sense and includes inclusion compounds without chemical bonds between water and the compound.

[0096] In the present application, a solvate refers to a higher-order compound formed between the molecules or ions of a solute and the molecules or ions of a solvent.

[0097] The extract of camellia oil meal contains terpene glycosides, flavonoid glycosides, sugars and proteins. In particular, the flavonoid glycosides in camellia oil meal include tetraglycosides (chemical formulas 1, 2, 3 and 4) and triglycosides (chemical formulas 5, 6, 7 and 8) of kaempferol.

[0098] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 1 is Kaempferol-3-O-β-D-xylopyranosyl-(1→3)-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-xylopyranosyl-(1→2)-O-β-D-galactopyranoside, which can exist in the form of a light yellow powder at room temperature.

[0099] Information about the kaempferol glycoside compound represented by Chemical Formula 1 is as follows.

[0100] Light yellow powder; UV (HPLC-PDA) λ max 266, 347 nm; [α] 22 D -57.4 (c 0.10); ESI-Q-TOF-MS: m / z 859.2503 [M+H] + (C 37 H 47 O 23 + Calculated value of, 859.2508), 881.2329 [M+Na] + (C 37 H 46 O 23 Na + Calculated value of, 859.2327); 1 H-(CD 3 OD, 500 MHz) and 13 C-NMR (CD 3 OD, 125Mhz) (see Figure 6 ).

[0101] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 2 is Kaempferol-3-O-β-D-xylopyranosyl-(1→3)-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-xylopyranosyl-(1→2)-O-β-D-glucopyranoside, which can exist in the form of a light yellow powder at room temperature.

[0102] Information on the kaempferol glycoside compound represented by Chemical Formula 2 is as follows.

[0103] Light yellow powder; UV (HPLC-PDA) λ max 266, 347 nm; [α] 22 D -63.7 (c 0.10); ESI-Q-TOF-MS: m / z 859.2505 [M+H] + (C 37 H 47 O 23 + calculated value of, 859.2508), 881.2325 [M+Na] + (C 37 H 46 O 23 Na + calculated value of, 859.2327); 1 H-(CD 3 OD, 500 MHz) and 13 C-NMR(CD 3 OD, 125Mhz) (see Figure 6 ).

[0104] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 3 is Kaempferol-3-O-β-D-xylopyranosyl-(1→3)-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-glucopyranosyl-(1→2)-O-β-D-galactopyranoside, which can exist in the form of a light yellow powder at room temperature.

[0105] Information on the kaempferol glycoside compound represented by Chemical Formula 3 is as follows.

[0106] Pale yellow powder; UV (HPLC - PDA) λ max 266, 347 nm; [α] 22 D -72.5 (c 0.2); ESI - Q - TOF - MS: m / z 889.2616 [M + H] + (C 38 H 49 O 23 + Calculated value of, 889.2613), and 911.2432 [M + Na] + (C 38 H 48 O 24 Na + Calculated value of, 911.2432); 1 H - (CD 3 OD, 500 MHz) and 13 C - NMR (CD 3 OD, 125 Mhz) (see Figure 6 ).

[0107] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 4 is Kaempferol - 3 - O - β - D - xylopyranosyl - (1→3) - O - α - L - rhamnopyranosyl - (1→6) - O - β - D - glucopyranosyl - (1→2) - O - β - D - gluctopyranoside, and it can exist in the form of pale yellow powder at room temperature.

[0108] Information on the kaempferol glycoside compound represented by Chemical Formula 4 is as follows.

[0109] Pale yellow powder; UV (HPLC - PDA) λ max 266, 347 nm; [α] 22 D -45.1 (c 0.10); ESI - Q - TOF - MS: m / z 889.2613 [M + H] + (C 38 H 49 O 23 +Calculated value, 889.2613) and 911.2423 [M+Na] + (C 38 H 48 O 24 Na + Calculated value, 859.2327); 1 H-(CD 3 OD, 500MHz) and 13 C-NMR(CD 3 OD, 125Mhz) (see Figure 6 ).

[0110] The composition according to one aspect of the present application contains kaempferol triglycoside compound or kaempferol tetraglycoside compound extracted from camellia oil meal discarded after extracting oil from camellia seeds as an active ingredient, and thus can exhibit significantly excellent cytoprotective effects compared with existing compositions containing kaempferol glycoside compound as an active ingredient.

[0111] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 5 is kaempferol-3-O-β-D-xylopyranosyl-(1→3)-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-galactopyranoside, which can exist in the form of a pale yellow powder at room temperature.

[0112] Information about the kaempferol glycoside compound represented by Chemical Formula 5 is as follows.

[0113] Pale yellow powder; UV(HPLC-PDA)λ max 265, 346nm; [α] 22 D -62.1(c 0.10); ESI-Q-TOF-MS: m / z 727.2904 [M+H] + (C 32 H 39 O 19 + Calculated value, 727.2086) and 749.1915 [M+Na] + (C 32 H 38 O 19 Na + Calculated value, 749.1905); 1 H-(CD 3 OD, 500MHz) and 13C-NMR(CD 3 OD, 125Mhz) (see Figure 7 ).

[0114] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 6 is Kaempferol-3-O-β-D-xylopyranosyl-(1→3)-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-glucopyranoside, and it can exist in the form of a pale yellow powder at room temperature.

[0115] Information on the kaempferol glycoside compound represented by Chemical Formula 6 is as follows.

[0116] Pale yellow powder; UV (HPLC-PDA) λ max 265, 346 nm; [α] 22 D 53.9 (c 0.10); ESI-Q-TOF-MS: m / z 727.2097 [M+H] + (C 32 H 39 O 19 + calculated value, 727.2686); 1 H-(CD 3 OD, 500MHz) and 13 C-NMR(CD 3 OD, 125Mhz) (see Figure 7 ).

[0117] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 7 is Kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-xylopyranosyl-(1→2)-O-β-D-galactopyranoside, and it can exist in the form of a pale yellow powder at room temperature.

[0118] Information on the kaempferol glycoside compound represented by Chemical Formula 7 is as follows.

[0119] Pale yellow powder; UV (HPLC-PDA) λ max 265, 346 nm; [α] 22D -70.8 (c 0.15); ESI-Q-TOF-MS: m / z 727.2108 [M+H] + (C 32 H 39 O 19 + Calculated values for (C + (C 32 H 38 O 19 Na + are 727.2086) and 749.1916 [M+Na] 1 H-(CD 3 OD, 500 MHz) and 13 C-NMR(CD 3 OD, 125 MHz) (see Figure 7 ).

[0120] The IUPAC name of the kaempferol glycoside compound represented by Chemical Formula 8 is Kaempferol-3-O-α-L-rhamnopyranosyl-(1→6)-O-β-D-xylopyranosyl-(1→2)-O-β-D-glucopyranoside, which can exist in the form of a pale yellow powder at room temperature.

[0121] Information about the kaempferol glycoside compound represented by Chemical Formula 8 is as follows.

[0122] Pale yellow powder; UV(HPLC-PDA) λ max 265, 346 nm; [α] 22 D -39.6 (c 0.10); ESI-Q-TOF-MS: m / z 749.1902 [M+Na] + (C 32 H 38 O 19 Na + Calculated values for (C 1 H-(CD 3 OD, 500 MHz) and 13 C-NMR(CD 3 OD, 125 MHz) (see Figure 7 ).

[0123] In one embodiment, the kaempferol glycoside compound can be extracted from camellia oil meal.

[0124] Specifically, the kaempferol glycoside compound can be obtained as follows: after extracting oil from camellia seeds by the pressing method, camellia oil meal is obtained, and then the kaempferol glycoside compound is extracted from the camellia oil meal using 70% ethanol.

[0125] In one embodiment, based on the total weight of the composition, the concentration of the active ingredient can be from 0.01 μM to 100 μM. For example, it can be above 0.01 μM, above 0.1 μM, above 0.5 μM, above 1 μM, above 1.5 μM, above 2 μM, or above 2.5 μM, and below 100 μM, below 50 μM, below 10 μM, below 8 μM, below 6 μM, or below 4 μM.

[0126] In one embodiment, the cell can be a skin cell.

[0127] In one embodiment, the composition can be used to inhibit or prevent skin damage.

[0128] In one embodiment, the composition can be used to improve skin self-renewal ability.

[0129] In one embodiment, the composition can inhibit the expression of DNA damage genes.

[0130] In one embodiment, the DNA damage gene can be γH2AX.

[0131] For example, when cells are irradiated with ultraviolet light, the DNA in the cell nucleus is damaged and will try to repair the damage itself. However, when the damage is too severe to be repaired, γH2AX will form at the Ser-139 residue of histone H2AX of DNA. Therefore, γH2AX is widely used as a well-known DNA damage marker. (See Figure 1 )

[0132] In one embodiment, the damage can be damage caused by ultraviolet light.

[0133] In one embodiment, the ultraviolet light can be at least one of UVA and UVB.

[0134] In one embodiment, the composition can be a cosmetic composition.

[0135] The form of the cosmetic composition according to an embodiment of the present application may include a cosmetically or dermatologically acceptable medium or matrix. It can be prepared into all dosage forms suitable for topical application. For example, it can be prepared into solutions, gels, solids, anhydrous pastes, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microsphere particles, or ionic (liposomes) and non-ionic vesicle dispersants, or in the form of creams, toners, lotions, powders, ointments, sprays or sticks. These compositions can be prepared by conventional methods in the art. The compositions according to the present invention can also be used in the form of foam or in the form of an aerosol composition further containing a compressed propellant.

[0136] The dosage form of the cosmetic composition according to an embodiment of the present invention is not particularly limited. For example, it can be prepared into cosmetics such as skin softening lotion, astringent lotion, nutritive lotion, nutritive cream, massage cream, essence, eye cream, eye essence, cleansing cream, cleansing foam, cleansing water, facial mask, powder, body lotion, body cream, body oil and body essence.

[0137] When the dosage form according to an embodiment of the present invention is a paste, cream or gel, animal fibers, plant fibers, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, polysiloxanes, bentonites, silica, talc or zinc oxide can be used as carrier components.

[0138] When the dosage form according to an embodiment of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder can be used as carrier components. In particular, when the dosage form is a spray, a propellant such as chlorofluorocarbon, propane / butane or dimethyl ether can be further included.

[0139] When the dosage form according to an embodiment of the present invention is a solution or an emulsion, solvents, solubilizers or emulsifying agents are used as carrier components. For example, water, ethanol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butanediol oil, glycerol fatty acid esters, polyethylene glycol or fatty acid esters of sorbitan anhydride.

[0140] When the dosage form according to an embodiment of the present invention is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitan esters and polyoxyethylene sorbitan anhydride esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth can be used as carrier components.

[0141] When the dosage form according to an embodiment of the present invention is a facial cleanser containing a surfactant, as the carrier component, fatty alcohol sulfates, fatty alcohol ether sulfates, sulfosuccinic acid monoesters, hydroxyethyl sulfonates, imidazoline derivatives, methyl taurates, sarcosinates, fatty acid amide ether sulfates, alkylamide betaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters, etc. can be used.

[0142] In a cosmetic composition according to an embodiment of the present invention, in addition to containing the above-mentioned active ingredient, functional additives and components contained in a normal cosmetic composition can also be included. The functional additives can include components selected from water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extracts.

[0143] Furthermore, in a cosmetic composition according to an embodiment of the present invention, in addition to the above-mentioned functional additives, components contained in a normal cosmetic composition can also be added as needed. In addition, the additional components can be oil components, moisturizers, emollients, surfactants, organic pigments and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, bactericides, antioxidants, plant extracts, pH regulators, alcohols, pigments, fragrances, blood circulation promoters, coolants, antiperspirants, pure water, etc.

[0144] In addition, the composition according to an embodiment of the present invention can be a topical skin agent, which is a general term for all dosage forms applied to the outside of the skin, and can include various dosage forms of cosmetics and pharmaceuticals.

[0145] In one embodiment, the composition can be a pharmaceutical composition.

[0146] In one embodiment, the composition can be a pharmaceutical composition for improving, preventing, or treating diseases caused by light stimulation.

[0147] The pharmaceutical composition according to an embodiment of the present invention can further contain preservatives, stabilizers, wettable powders or emulsification promoters, pharmaceutical adjuvants for adjusting osmotic pressure such as salts and / or buffers, and other substances useful for treatment, and can be formulated into various oral dosage forms or parenteral dosage forms according to conventional methods.

[0148] The oral dosage forms may be, for example, tablets, pills, hard capsules and soft capsules, liquids, suspensions, emulsions, syrups, powders, powders, fine granules, granules and pellets, etc. In addition to the active ingredient, these dosage forms may also contain surfactants, 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 glycol). In addition, tablets may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose and polyvinylpyrrolidone, and may also contain disintegrants, absorbents, coloring agents, flavoring agents, and sweetening agents such as starch, agar, alginic acid or its sodium salt, etc. according to specific circumstances. The tablets can be prepared by conventional mixing, granulation or coating methods. In addition, the parenteral dosage forms may be percutaneous dosage forms, such as injection, drops, ointment, lotion, gel, cream, spray, suspension, emulsion, suppository, patch and other dosage forms, but not limited thereto.

[0149] The pharmaceutical composition according to an embodiment of the present invention can be administered parenterally, rectally, topically, percutaneously, subcutaneously, etc.

[0150] Those skilled in the art can determine the dosage of the active ingredient based on their cognitive level. Although the daily dosage of the drug varies according to various factors such as the disease progression degree, onset period, age, health status, and complications of the subject to be administered, generally, based on adults, the dosage of the composition can be 0.01 mg / kg / day to 100 mg / kg / day. For example, it can be above 0.01 mg / kg / day, above 0.1 mg / kg / day, above 0.5 mg / kg / day, above 1 mg / kg / day, above 1.3 mg / kg / day, above 1.5 mg / kg / day, above 1.7 mg / kg / day, above 2 mg / kg / day, or above 2.2 mg / kg / day, and below 100 mg / kg / day, below 50 mg / kg / day, below 10 mg / kg / day, below 7 mg / kg / day, below 5 mg / kg / day, below 3 mg / kg / day, or below 2.5 mg / kg / day. Preferably, 1 mg / kg to 5 mg / kg can be divided into 1 to 3 administrations per day, and this dosage does not limit the scope of the present invention in any way.

[0151] In one embodiment, the composition may be an oral composition.

[0152] The oral composition according to an embodiment of the present invention may be in a liquid or solid dosage form, and may be a tablet, capsule, soft capsule, pill, granule, beverage (drink), weight loss bar, chocolate, caramel dosage form or biscuit dosage form, and its dosage form is not particularly limited. In addition to the active ingredient, the oral composition of the present invention may optionally contain excipients, sugars, flavors, pigments, oils and fats, proteins, etc. in appropriate amounts.

[0153] Hereinafter, the present invention will be described more specifically by the following examples. However, the purpose of providing these examples is only to help understand the present invention, and the scope and range of the present invention are not limited by them.

[0154]

Example

[0155] The camellia oil meal used in the experiment was obtained by pressing camellia seeds in the camellia village of Jeju Island to produce oil. An extract was prepared from the camellia oil meal using 70% ethanol, and it was confirmed that the camellia oil meal extract contained terpene glycosides, flavonoid glycosides, sugars and proteins. In particular, the flavonoid glycosides of the camellia oil meal include tetraglycosides (chemical formulas 1 to 4) and triglycosides (chemical formulas 5 to 8) of kaempferol.

[0156] Purification of kaempferol glycosides from camellia oil meal

[0157] 8 kg of 70% ethanol was added to 1 kg of defatted camellia oil meal after low-temperature oil pressing, and extraction was carried out at 60 °C for 8 hours using a heating extractor. The extract obtained by filtration through a pressure filter was concentrated under reduced pressure using a rotary vacuum concentrator at 50 °C to 60 °C until the solid content was about 50%, thereby obtaining 200 g of concentrated solution. The concentrated solution was diluted with pure water to a solid content of 10%.

[0158] After loading this solution onto a glass column filled with 2 L of ion exchange resin (HP-20), it was washed with 10 L of pure water to remove impurities and sugar components. Then 10 L of 30% ethanol was passed through in sequence, and the eluate was concentrated under reduced pressure and freeze-dried to obtain about 20 g of camellia kaempferol glycoside powder.

[0159] That is, the finally obtained camellia kaempferol glycoside powder is in a form containing chemical formulas 1 to 4 (tetraglycosides) and chemical formulas 5 to 8 (triglycosides).

[0160]

Experimental Example

[0161]

Experimental Example 1

[0162] HaCaT cells were cultured in DMEM medium supplemented with 10% FBS and 1% PS. For immunofluorescence staining, cells were seeded at 50,000 cells / well onto 8-well chamber slides. The next day, cells were pretreated with kaempferol glycoside at the following concentrations for 1 hour. The treatment concentrations were 2.5 ppm and 5.10 ppm. After 1 hour of pretreatment, the cells were washed with PBS and then irradiated with 15 mJ / cm2 of UVB. At this time, the control group was not irradiated with UVB.

[0163] The experiment was conducted by dividing the cells into a group not irradiated with UVB (cont), a group irradiated only with UVB, and a group post-treated with kaempferol glycoside in the same manner as the pretreatment conditions after UVB irradiation. After a total of 24 hours, the cells were collected for immunofluorescence staining.

[0164] For immunofluorescence staining, cells in each well were washed with PBS containing 1 mM of CaCl2 and 1 mM of MgCl2. The cells were reacted and fixed with 3.5% paraformaldehyde at room temperature for 10 minutes. The fixed cells were washed again with PBS, 3 times for 10 minutes each. The cells were treated with 0.1% Triton X-100 for 5 minutes. The cells were washed again with PBS, 3 times for 10 minutes each. The p-H2AX antibody (Cell Signaling) was diluted 1:100 in PBS containing 0.05% Tween 20 (PBST) and reacted with the primary antibody at 4 °C O / N. The cells were washed again with PBST, 3 times for 10 minutes each, and then reacted with a rhodamine-labeled secondary antibody at 1:200 at room temperature for 1 hour. After that, the cells were washed with PBST, 3 times for 10 minutes each. To stain the nuclei, propidium iodide (PI) was used for staining for about 3 minutes. The cells were washed 3 times with PBST again, and then a mounting solution was added and a coverslip was covered. The stained slides were photographed using a confocal laser scanning microscope (Ziess). Each image was photographed at least 6 times.

[0165] Using the confocal dedicated program ZEN, the intensity of each fluorescence of each photographed image was measured. The results are shown respectively as Figure 2 and Figure 3 shown. Specifically, since all fluorescence values are the fluorescence counts of individual cells, the total red signal in one image was measured, and then the number of cells stained with DAPI was measured and calculated, as Figure 3 shown. (n = 6 or more)

[0166] Figure 2The upper part only shows red signals, and the result can be qualitatively confirmed that the expression of γH2AX (red signal), a marker of DNA damage caused by UVB, is strong, and it is inhibited in a concentration-dependent manner by kaempferol glycoside. In addition, as Figure 2 shown in the lower part, it can be qualitatively confirmed that γH2AX (red signal), a marker of DNA damage caused by UVB, is strongly expressed in the nucleus (the same position as the blue signal) (red-pink), but after being inhibited in a concentration-dependent manner by kaempferol glycoside, the color gets closer and closer to blue in the right direction, and it is confirmed by measuring the intensity of γH2AX (red signal) that its value gets smaller and smaller.

[0167]

Experimental Example 2

[0168] Ultraviolet light can cause cell death and growth reduction, and it is confirmed that the cell protection effect is achieved by overcoming this situation with kaempferol glycoside. Specifically, in order to confirm cell viability under ultraviolet light, the cell number was directly measured. The DAPI images stained in Experimental Example 1 above were analyzed separately to measure the cell number. The results are as Figure 4 and Figure 5 shown.

[0169] As Figure 4 shown, the nuclei of living cells were stained with DAPI (blue signal), and the cell number could be directly confirmed with the naked eye. It was confirmed that UVB caused a decrease in the cell number, and kaempferol glycoside had the effect of protecting the cell number under ultraviolet light. From the quantitative analysis results of the cell number ( Figure 5 ), it can be seen that the cell number increased significantly. Therefore, the effect of directly improving cell viability under UVB was confirmed.

[0170]

Embodiment

[0171] Embodiment 1: Use of a kaempferol glycoside compound, its isomer, its pharmaceutically acceptable salt, its hydrate or its solvate in the preparation of a composition for repairing cell DNA damage, wherein the kaempferol glycoside compound is represented by any one of the following Chemical Formulas 1 to 8,

[0172]

Chemical Formula 1

[0173]

[0174]

Chemical Formula 2

[0175]

[0176]

Chemical Formula 3

[0177]

[0178]

Chemical Formula 4

[0179]

[0180]

Chemical Formula 5

[0181]

[0182]

Chemical Formula 6

[0183]

[0184]

Chemical Formula 7

[0185]

[0186]

Chemical Formula 8

[0187]

[0188] Embodiment 2: The use according to Embodiment 1, wherein the kaempferol glycoside compound is extracted from camellia oil meal.

[0189] Embodiment 3: The use according to Embodiment 1 or 2, wherein, based on the total weight of the composition, the concentration of the kaempferol glycoside compound, its isomer, its pharmaceutically acceptable salt, its hydrate or its solvate represented by any one of Chemical Formulas 1 to 8 is 0.01 μM to 100 μM.

[0190] Embodiment 4: The use according to any one of Embodiments 1 to 3, wherein the cell is a skin cell.

[0191] Embodiment 5: The use according to any one of Embodiments 1 to 4, wherein the composition is used for inhibiting or preventing skin damage.

[0192] Embodiment 6: The use according to any one of Embodiments 1 to 5, wherein the composition is used for improving skin self-renewal ability.

[0193] Embodiment 7: The use according to any one of Embodiments 1 to 6, wherein the composition inhibits the expression of DNA damage genes.

[0194] Embodiment 8: The use according to any one of Embodiments 1 to 7, wherein the DNA damage gene is γH2AX.

[0195] Embodiment 9: The use according to any one of Embodiments 1 to 8, wherein the damage is damage caused by ultraviolet rays.

[0196] Embodiment 10: The use according to any one of Embodiments 1 to 9, wherein the ultraviolet ray is at least one of UVA and UVB.

[0197] Embodiment 11: The use according to any one of Embodiments 1 to 10, wherein the composition is a cosmetic composition.

[0198] Embodiment 12: The use according to any one of Embodiments 1 to 11, wherein the composition is a pharmaceutical composition.

[0199] Embodiment 13: The use according to any one of Embodiments 1 to 12, wherein the application amount of the active ingredient is 0.01 mg / kg / day to 100 mg / kg / day.

Claims

1. Use of a kaempferol glycoside compound, its isomers, its pharmaceutically acceptable salts, its hydrates or its solvates in preparing a composition for repairing cell DNA damage, wherein: The kaempferol glycoside compound is represented by any one of the following chemical formulas 1 to 8, 【Chemical formula 1】 【Chemical formula 2】 【Chemical formula 3】 【Chemical formula 4】 【Chemical formula 5】 【Chemical formula 6】 【Chemical formula 7】 【Chemical formula 8】 2. The use according to claim 1, wherein The kaempferol glycoside compound is extracted from camellia oil cake.

3. The use according to claim 1, wherein The concentration of the kaempferol glycoside compound, its isomer, its pharmaceutically acceptable salt, its hydrate or its solvate is 0.01 μM to 100 μM based on the total weight of the composition.

4. The use according to claim 1, wherein The cells are skin cells.

5. The use according to claim 1, wherein The composition is used to inhibit or prevent skin damage.

6. The use according to claim 1, wherein The composition is used to improve the skin's self-regeneration ability.

7. The use according to claim 1, wherein The composition inhibits the expression of DNA damaging genes.

8. The use according to claim 1, wherein The DNA damage gene is γH2AX.

9. The use according to claim 1, wherein The damage is damage caused by ultraviolet rays.

10. The use according to claim 9, wherein The ultraviolet rays are at least one of UVA and UVB.

11. The use according to claim 1, wherein The composition is a cosmetic composition.

12. The use according to claim 1, wherein The composition is a pharmaceutical composition.

13. The use according to claim 12, wherein The administration amount of the kaempferol glycoside compound, its isomer, its pharmaceutically acceptable salt, its hydrate or its solvate is 0.01 mg / kg / day to 100 mg / kg / day.

Images