Composition comprising novel ginsenoside for protecting skin or respiratory against finedust

Abstract

This specification relates to a composition comprising, as an active ingredient, a novel ginsenoside (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranosyl-dammar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof. The composition exhibits excellent skin or respiratory protective effects against fine dust.

Description

Technology Field

[0001] This specification describes novel ginsenosides and compositions containing them. Background Technology

[0002] Ginseng (*Panax ginseng CA Meyer*) is a plant belonging to the genus Panax in the family Araliaceae and is a medicinal herb that has been used in Korea, China, Japan, and other regions for over 2,000 years. Saponins, polysaccharides, peptides, sitosterol, polyacetellin, and fatty acids are known as the representative physiologically active components of ginseng; among these, the saponins of ginseng are called ginsenosides. The efficacy and effects of ginseng are known to include actions on the central nervous system, anti-carcinogenic and anticancer activity, immune function regulation, anti-diabetic activity, liver function enhancement, improvement of cardiovascular disorders and anti-atherosclerosis, blood pressure regulation, improvement of menopausal disorders and effects on osteoporosis, anti-stress and anti-fatigue activity, antioxidant activity, and anti-aging efficacy. Although the content and composition of the aforementioned ginsenosides vary significantly depending on the part of the ginseng plant, such as the root, leaves, fruit, flower, and seeds, the known efficacy described above is mostly attributed to the ginseng root, and research on other parts of the ginseng plant excluding the root is currently lacking. Prior art literature

[0003] Republic of Korea Published Patent Application No. 10-2016-0086149 The problem to be solved

[0004] In one aspect, the problem that the present invention aims to solve is to provide a composition comprising a novel ginsenoside having skin or respiratory protective efficacy against fine dust. means of solving the problem

[0005] In one aspect, the present invention provides a composition for skin or respiratory protection against fine dust comprising (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient. Effects of the invention

[0006] In one aspect, the present invention may provide a composition comprising a novel ginsenoside having an excellent effect on skin protection or respiratory protection against fine dust, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof. Brief explanation of the drawing

[0007] Figure 1 is a figure illustrating the isolation process of the novel ginsenoside (Cpd. 10) of the present invention among compounds fractionated from ginseng seed extract. Figure 2 shows the chemical structures of 16 compounds fractionated from ginseng seed extract. Figure 3 is a figure showing the spectroscopic evidence and structures of compounds 1 to 6, which correspond to previously known ginsenosides among the compounds fractionated from ginseng seed extract. FIG. 4 shows compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. 1 This is a diagram showing the H-NMR spectrum. FIG. 5 shows compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. 13 This is a diagram showing the C-NMR spectrum. Figure 6 is a figure showing the COSY spectrum of compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. Figure 7 is a figure showing the HSQC spectrum of compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. Figure 8 is a figure showing the HMBC spectrum of compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. Figure 9 is a figure showing the MS spectrum of compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. Figure 10 is a figure showing the core HMBC correlation of compound 10, which corresponds to the novel ginsenoside of the present invention among the compounds fractionated from ginseng seed extract. Figure 11 shows the efficacy of compound 10 (GS#10), corresponding to the novel ginsenoside of the present invention, in preventing skin cell death caused by fine dust. (*** P < 0.001 vs. PM10, ** P < 0.01 vs. PM10, * P < 0.05 vs. PM10) Figure 12 shows a comparison of the efficacy of ginseng seed extract (GSE) and compound 10 (GS#10), which corresponds to the novel ginsenoside of the present invention, in preventing skin cell death caused by fine dust. ( *** P < 0.001 vs. PM10, * P < 0.05 vs. PM10) Figure 13 shows a comparison of the efficacy of ginsenosides Rg1, Rg3, and Rb1, which are indicator components of red ginseng, and compound 10 (GS#10), which corresponds to the novel ginsenoside of the present invention, in preventing skin cell death caused by fine dust. (*** P < 0.001 vs. PM10, * P < 0.05 vs. PM10) Figure 14 shows the efficacy of compound 10 (GS#10), corresponding to the novel ginsenoside of the present invention, in preventing lung cell death caused by fine dust. (*** P < 0.001 vs. PM10, ** P < 0.01 vs. PM10, * P < 0.05 vs. PM10) Figure 15 shows a comparison of the efficacy of ginseng seed extract (GSE) and compound 10 (GS#10), which corresponds to the novel ginsenoside of the present invention, in preventing lung cell death caused by fine dust. ( *** P < 0.001 vs. PM10, ** P < 0.01 vs. PM10, * P < 0.05 vs. PM10) Figure 16 shows a comparison of the efficacy of ginsenosides Rg1, Rg3, and Rb1, which are indicator components of red ginseng, and compound 10 (GS#10), which corresponds to the novel ginsenoside of the present invention, in preventing lung cell damage caused by fine dust. (** P < 0.01 vs. UVB, * P < 0.05 vs. UVB) Figure 17 shows the cell viability (%Viable cells) of Compound 10 (GS#10), which corresponds to the novel ginsenoside of the present invention. (*** P < 0.001 vs. (-), ** P < 0.01 vs. (-), * P < 0.05 vs. (-)) Specific details for implementing the invention

[0008] Hereinafter, embodiments of the present application will be described in more detail with reference to the attached drawings. However, the technology disclosed in the present application is not limited to the embodiments described herein and may be embodied in other forms. The embodiments introduced herein are provided merely to ensure that the disclosed content is thorough and complete and to ensure that the concept of the present application is sufficiently conveyed to those skilled in the art. In the drawings, the size of the components, such as their width or thickness, has been shown slightly enlarged to clearly represent each component. Additionally, for convenience of explanation, only parts of the components have been illustrated, but those skilled in the art will be able to easily understand the remaining parts of the components. Furthermore, those with ordinary knowledge in the relevant field may embody the concept of the present application in various other forms without departing from the technical concept of the present application.

[0010] In one embodiment, the present invention may provide a composition for protecting the skin or respiratory tract against fine dust, comprising a novel ginsenoside, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

[0011] As used in this specification, "fine dust" refers to particulate matter that floats or is scattered in the atmosphere for a long time. Dust is classified according to particle size into Total Suspended Particles (TSP), which are 50 μm or smaller, and Particulate Matter (PM), which are very small particles. The components of fine dust may vary depending on the region, season, and weather conditions where the fine dust is generated, and may include, for example, clusters formed by the reaction of air pollutants in the air (such as sulfates and nitrates), carbon compounds and soot generated during the burning of fossil fuels such as coal and petroleum, and minerals generated from surface soil dust. According to the World Health Organization (WHO), fine dust is classified into fine dust with a diameter of 10 μm or less (PM10) and small fine dust with a diameter of 2.5 μm or less, i.e., ultrafine dust (PM2.5). Fine dust (PM10) with a diameter of 10 μm or less may include, for example, soil dust, pollen, mold, etc. Ultrafine dust (PM2.5) with a diameter of 2.5 μm or less may include, for example, combustion particles, organic compounds, metals, etc. Because fine dust is so small that it is invisible to the naked eye, it remains in the atmosphere and can have adverse effects on health by penetrating the lungs through the respiratory system or traveling into the body via blood vessels. The International Agency for Research on Cancer (IARC), under the World Health Organization, has designated fine dust as a Group 1 carcinogen, meaning it is confirmed to be carcinogenic to humans. In this invention, the term “fine dust” is used in the broadest sense to include all substances capable of affecting the human body, without being limited by the size or composition of the dust.

[0012] In one embodiment, the ginsenoside is a novel triterpene saponin, (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol.

[0013] One embodiment may provide for the use of (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-damar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof for use in the manufacture of compositions for skin or respiratory protection against fine dust.

[0014] One embodiment may provide a method for protecting the skin or respiratory tract against fine dust, comprising administering an effective amount of (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-damar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof to a subject in need thereof.

[0015] One embodiment may provide (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranosyl-damar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient for use in a composition for skin or respiratory protection against fine dust. Additionally, non-therapeutic use of (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranosyl-damar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient for skin or respiratory protection against fine dust may be provided.

[0016] In this specification, “pharmaceuticalally acceptable” means that it is approved by a government or equivalent regulatory authority for use in animals, more specifically in humans, by avoiding significant toxic effects when used at a normal medicinal dosage, or is recognized as being listed in a pharmacopoeia or described in other general pharmacopoeias.

[0017] In this specification, "pharmaceuticalally acceptable salt" means a salt according to one aspect of the present invention that is pharmaceutically acceptable and has the desirable pharmacological activity of a parent compound. The salt is formed from (1) an inorganic acid such as hydrochloric acid, hydrobromide, sulfuric acid, nitric acid, phosphoric acid, etc.; or acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvate, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2,2,2]-oct-2-en-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, It may include an acid addition salt formed from organic acids such as tert-butylacetic acid, lauryl sulfate, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, and muconic acid; or (2) a salt formed when an acidic proton present in the parent compound is substituted.

[0018] In this specification, "hydrate" refers to a compound to which water is bound, and is a broad concept that includes inclusion compounds in which there is no chemical bonding force between the water and the compound.

[0019] In this specification, "solvent" refers to a higher-order compound formed between molecules or ions of a solute and molecules or ions of a solvent.

[0020] In one embodiment, the molecular formula of the ginsenoside is C 42 H 70 O 15 It is and has the following chemical structure.

[0021] [Chemical Formula 1]

[0022]

[0023] In this specification, the novel ginsenoside is named "pseudoginsenoside RT8" or "PG-RT8".

[0024] In one embodiment, the ginsenoside may be extracted from ginseng seeds. More specifically, the ginsenoside may be isolated from a ginseng seed extract, but is not limited thereto. In one embodiment, the ginseng of the ginseng seeds is Panax ginseng ( Panax ginseng CA Meyer)

[0025] In this specification, the term "separation" means including extraction or fractionation from ginseng seed extract, and may involve the use of water, organic solvents, etc., and any method known to those skilled in the art may be applied. The fractionation may be performed after the extraction.

[0026] In this specification, the term "extract" includes all substances obtained by extracting components from a natural product, regardless of the extraction method or the type of component. For example, it is a broad concept that includes substances obtained by extracting solvent-soluble components from a natural product using water or an organic solvent, and substances obtained by extracting only specific components of a natural product.

[0027] In this specification, "fraction" includes fractions of a specific substance or extract obtained by fractionating it using a specific solvent, the remainder after fractionation, and the fractions obtained by re-extracting them with a specific solvent. Any method known to a person skilled in the art may be used for fractionation and extraction.

[0028] In one embodiment, the ginsenoside may be isolated from the methanol and butanol-soluble extract of ginseng seeds. Specifically, the ginsenoside may be detected and isolated by analyzing the methanol and butanol-soluble extract of ginseng seeds using HPLC-ESI-Q-TOF-MS. Since the main component of the ginseng seed extract is lipids, not all triterpenes and steroidal saponins can be observed from the crude ginseng seed extract by HPLC-UV or HPLC-ELSD.

[0029] In one embodiment, the present invention may provide a composition that inhibits damage or death of skin cells caused by fine dust by including the ginsenoside, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof. For example, one embodiment of the present invention may prevent, improve, or treat skin irritation or skin diseases caused by fine dust. In one embodiment, the skin diseases caused by fine dust may include inflammatory skin diseases such as contact dermatitis, atopic dermatitis, allergic dermatitis, acne, etc.

[0030] In one embodiment, the present invention may provide a composition that inhibits damage or death of lung cells caused by fine dust by comprising the ginsenoside, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof. For example, one embodiment of the present invention may prevent, improve, or treat respiratory diseases caused by fine dust. In this specification, the term “respiratory disease” means a disease occurring in the respiratory organs, including the nose, airways, bronchi, or lungs. In one embodiment, the respiratory disease may include, for example, chronic obstructive pulmonary diseases such as asthma, emphysema, or chronic bronchitis, lung cancer, etc.

[0031] In this specification, the term “prevention” refers to any act of suppressing or delaying a desired symptom by administering a composition according to one embodiment of the present invention. In this specification, the term “treatment” refers to any act of improving or causing a desired symptom or disease to disappear by administering a composition according to one embodiment of the present invention. In this specification, the term “improvement” refers to any act of improving or beneficially changing a desired symptom compared to before administration by administering a composition according to one embodiment of the present invention.

[0032] In one embodiment, the present invention may include the active ingredient in an amount of 0.0001 to 99.9 weight% based on the total weight of the composition. Specifically, as one embodiment, the above composition comprises the active ingredient in an amount of 0.0001 wt% or more, 0.0005 wt% or more, 0.001 wt% or more, 0.01 wt% or more, 0.1 wt% or more, 1 wt% or more, 2 wt% or more, 3 wt% or more, 4 wt% or more, 5 wt% or more, 6 wt% or more, 7 wt% or more, 8 wt% or more, 9 wt% or more, 10 wt% or more, 15 wt% or more, 20 wt% or more, 25 wt% or more, 30 wt% or more, 35 wt% or more, 40 wt% or more, 45 wt% or more, 50 wt% or more, 55 wt% or more, 60 wt% or more, 65 wt% or more, 70 wt% or more, 75 wt% or more, 80 wt% or more, 85 wt% or more, 90 wt% or more, 95 wt% or more, or based on the total weight of the composition. It may contain 99.9% by weight or more, but is not limited to the above range. Or, as one embodiment, the above composition comprises the active ingredient in an amount of 100 wt% or less, 99 wt% or less, 95 wt% or less, 90 wt% or less, 85 wt% or less, 80 wt% or less, 75 wt% or less, 70 wt% or less, 65 wt% or less, 60 wt% or less, 55 wt% or less, 50 wt% or less, 45 wt% or less, 40 wt% or less, 35 wt% or less, 30 wt% or less, 25 wt% or less, 20 wt% or less, 15 wt% or less, 10 wt% or less, 9 wt% or less, 8 wt% or less, 7 wt% or less, 6 wt% or less, 5 wt% or less, 4 wt% or less, 3 wt% or less, 2 wt% or less, 1 wt% or less, 0.5 wt% or less, 0.1 wt% or less, 0.01 wt% or less, 0.001 wt% or less, or based on the total weight of the composition. It may be included in an amount of 0.0005 weight% or less, but is not limited to the above range.

[0033] In one embodiment, the dosage of the composition may be from 0.05 mg / kg / day to 10 g / kg / day. In one embodiment, the dosage of the composition may vary depending on the subject's age, gender, body weight, specific disease or pathological condition, severity of the disease or pathological condition, route of administration, etc., and the determination of the dosage based on these factors is within the level of a person skilled in the art. For example, the dosage may be 0.05 mg / kg / day or 1 mg / kg / day or more, and 10 g / kg / day or less, 100 mg / kg / day or less, or 10 mg / kg / day or less, but the dosage does not limit the scope of this specification in any way.

[0034] In one embodiment, the dosage of the composition may be from 0.1 μg / mL / day to 10 mg / mL / day. In one embodiment, the dosage of the composition may vary depending on the subject's age, gender, body weight, specific disease or pathological condition, severity of the disease or pathological condition, route of administration, etc., and the determination of the dosage based on these factors is within the level of a person skilled in the art. For example, the dosage may be 0.1 μg / mL / day or more or 1 μg / kg / day or more, and may be 10 mg / mL / day or less, 1 mg / L / day or less, or 100 μg / L / day or less, but the dosage does not limit the scope of this specification in any way.

[0035] The composition according to the embodiments of the present invention may be a skin external preparation composition comprising the above active ingredient.

[0036] In this specification, the term "skin" refers to tissue covering the surface of an animal's body, and is a broad concept that includes not only tissue covering the surface of the body such as the face or body, but also the scalp and hair.

[0037] The composition according to the embodiments of the present invention may be a cosmetic composition comprising the active ingredient.

[0038] In one embodiment, the composition may be formulated to contain a cosmetically or dermatologically acceptable medium or base. This may be provided in any formulation suitable for topical application, for example, in the form of a solution, gel, solid, paste anhydrous product, emulsion obtained by dispersing an oil phase in an aqueous phase, suspension, microemulsion, microcapsule, microgranule, or ionic (liposome) and non-ionic vesicular dispersant, or in the form of a cream, skin toner, lotion, powder, ointment, spray, or concealer stick. It may also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions may be prepared according to conventional methods in the art.

[0039] The composition according to the embodiments of the present invention may be a food composition comprising the active ingredient.

[0040] For example, it may be processed into functional foods such as fermented milk, cheese, yogurt, juice, probiotic preparations, and health foods containing the above active ingredient, and may also be used in the form of various other food additives. In one embodiment, the composition may be a composition for health foods. In one embodiment, the composition for health foods may be formulated into pills, capsules, tablets, granules, caramels, or drinks. In another embodiment, it may be processed into the form of liquids, powders, granules, tablets, or tea bags. The composition may be administered by various methods such as simple drinking, injection, spraying, or squeezing. The composition may contain other ingredients that can provide a synergistic effect to the main effect within a range that does not impair the main effect of the present invention. For example, to improve physical properties, it may further include additives such as flavorings, colorings, disinfectants, antioxidants, preservatives, humectants, thickeners, inorganic salts, emulsifiers, and synthetic polymer materials. In addition, auxiliary ingredients such as water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, and seaweed extract may be further included. The above ingredients may be appropriately selected and combined by a person skilled in the art according to the formulation or purpose of use, and the amount added may be selected within a range that does not impair the purpose and effect of the present invention. For example, the amount added of the above ingredients may be 0.0001% by weight to 99.9% by weight based on the total weight of the composition. As an example, the dosage of the food composition may vary depending on the subject's age, gender, body weight, specific disease or pathological condition, severity of the disease or pathological condition, route of administration, etc., and the determination of the dosage based on these factors is within the level of a person skilled in the art. For example, the above dosage is 0.It may be 0.5 mg / kg / day or more or 1 mg / kg / day or more, and may be 10 g / kg / day or less, 100 mg / kg / day or less, or 10 mg / kg / day or less, but the above dosage does not limit the range of this specification in any way.

[0041] The composition according to the embodiments of the present invention may be a pharmaceutical composition comprising the active ingredient. The pharmaceutical composition may further contain pharmaceutical adjuvants such as preservatives, stabilizers, wettable agents or emulsification promoters, salts and / or buffers for osmotic pressure regulation, and other therapeutically useful substances.

[0042] In one embodiment, the pharmaceutical composition may be an oral formulation, and the oral formulation includes, for example, tablets, pills, hard and soft capsules, liquids, suspensions, emulsifiers, syrups, powders, granules, granules, pellets, etc. These formulations may contain, in addition to the active ingredient, surfactants, diluents (e.g., lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, and glycine), and lubricants (e.g., silica, talc, stearic acid and its magnesium or calcium salts, and polyethylene glycol). The tablet may also contain binders such as magnesium aluminum silicate, starch paste, gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose, and polyvinylpyrrolidine, and, in some cases, may contain pharmaceutical additives such as disintegrants, absorbents, coloring agents, flavoring agents, and sweeteners, such as starch, agar, alginic acid, or its sodium salt. The tablet may be manufactured by conventional mixing, granulation, or coating methods.

[0043] In one embodiment, the pharmaceutical composition may be a parenteral agent, and the parenteral agent may be a rectal, topical, subcutaneous, or transdermal formulation. For example, it may be a formulation such as an injection, drops, ointment, lotion, gel, cream, spray, suspension, emulsion, suppository, or patch, but is not limited thereto.

[0044] In one embodiment, the dosage of the pharmaceutical composition will vary depending on the age, gender, and weight of the subject to treatment, the specific disease or pathological condition to be treated, the severity of the disease or pathological condition, the route of administration, and the judgment of the prescriber. The determination of the dosage based on these factors is within the level of a person skilled in the art. For example, the dosage may be 0.05 mg / kg / day or 1 mg / kg / day or more, and may be 10 g / kg / day or less, 100 mg / kg / day or less, or 10 mg / kg / day or less, but the dosage does not limit the scope of this specification in any way.

[0046] The present invention will be described in detail below with reference to examples, comparative examples, and test examples. These are presented merely as examples to explain the invention more specifically, and it will be obvious to those skilled in the art that the scope of the invention is not limited by these examples, comparative examples, and test examples.

[0047] All experimental values ​​below represent the average of values ​​from at least three repeated experiments, and the standard deviation (SD) is indicated by error bars. P-values ​​were calculated using one-way ANOVA and the Dunnett test, and p-values ​​less than 0.05 were considered statistically significant.

[0049] [Example 1] Separation of Ginsenosides

[0050] fraction

[0051] Ginseng seeds (Seeds of Panax ginseng5.5 kg was finely ground into a powder using a mixer, extracted with methanol, and then fractionated stepwise using n-hexane, ethyl acetate, and n-butanol. Most of the lipids were removed by n-hexane, and the lipids remaining in the ethyl acetate fraction were suspended in a methanol:water ratio of 1:1 (v / v). After storing the mixture in a freezer overnight, only the supernatant was collected and further removed using a centrifuge. The 2.61 g of the ethyl acetate fraction and 114.64 g of the n-butanol fraction obtained in this way were fractionated as follows using a column and HPCCC (High Performance Counter-Current Chromatography).

[0052] Column and HPCCC of n-butanol fractions

[0053] 114.64 g of the n-butanol fraction was divided into fractions using MPLC. The solvents used were n-hexane / ethyl acetate = 10:1 -> 5:1 -> 1:1 -> CHCl3 / MeOH = 10:1 -> 5:1 (v / v), and the flow rate was 50 mL / min. Using these conditions, the fraction was divided into a total of 12 sub-fractions. The components contained in each fraction were separated using HPCCC, HPLC (High-performance liquid chromatography), and a Sephadex LH-20 column. Subsequently, 16 compounds were identified by structural analysis using NMR (Nuclear magnetic resonance), UV (Ultraviolet rays), and MS (Mass spectrometry).

[0054] The above-mentioned isolated 16 compounds include protopanaxatriol saponins ginsenoside Rg1 (Compound 1), ginsenoside Rg2 (Compound 2), and ginsenoside Re (Compound 3); protopanaxadiol saponins ginsenoside Rd (Compound 4), ginsenoside Rb1 (Compound 5), and ginsenoside Rb2 (Compound 6); Stigma-5-en-3-O-β-D-glucopyranoside (Compound 7), Stigma-5,24(28)-dien-3-O-β-D-glucopyranoside (Compound 8), and Stigma-5,22-dien-3-O-β-D-glucopyranoside (Compound 9); (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol (Compound 10), a novel compound that is isolated for the first time from nature as a novel ginsenoside according to one embodiment of the present invention; phenolic glycosides phenethyl alcohol β-D-xylopyranosyl(1->6)-β-D-glucopyranoside (Compound 12) and eugenyl β-gentiobioside (Compound 13); the flavonoid isorhamnetin 3- O -β-D-glucopyranoside(isorhamnetin 3- O-β-D-glucopyranoside) (Compound 15); the primary metabolites were adenosine (Compound 11), uracil (Compound 14) and tryptophan (Compound 16).

[0055] The isolation process of a novel ginsenoside according to one embodiment of the present invention corresponding to the above compound 10 is illustrated in FIG. 1. The chemical structures of the above 16 types of compounds are illustrated in FIG. 2, and the spectroscopic evidence and chemical structures of compounds 1-6, which are previously known ginsenosides among the above compounds, are separately illustrated in FIG. 3.

[0056] Compound 10 in the cation ESI-Q-TOF-MS (Electrospray Ionization-Quadrupole-Time-of-flight mass spectrometry) spectrum m / z 837.4617 [(M+Na) + Based on the sodiated pseudomolecular ion peak in [calcd.837.4612], C 42 H 70 O 15 It was isolated as a white amorphous powder exhibiting the molecular formula of the above compound 10. 1 The H NMR spectrum is [δ H It contained 8 methyl resonances at [1.86(3H,s,H-28), 1.69(3H,s,H-29), 1.47(3H,s,H-27), 1.25(6H,s,H-21,26), 1.10(3H,s,H-18), 0.81(3H,s,H-30), 0.75(3H,s,H-19)]. Additionally, two pairs of signals corresponding to anomeric protons and carbon atoms at the two sugar residues were δ H 6.02(1H,d, J = 7.8, H-2") / δ C 104.08(C-1') and H 4.91 (1H, d, J= 7.7, H-1') / δ C It was detected in 104.32 (C-1). 13 CNMR and heteronuclear single quantum correlation (HSQC) spectra revealed 42-carbon signals. Apart from the two sugar residues mentioned above, the aglycone of compound 10 consists of 8 methylene, 4 methine, and 3 oxygen-containing methine [δ C 79.79 (C-6), 71.40 (C-12) and 86.09 (C-24)], 5 quaternary carbon atoms, 2 oxygenated quaternary carbon atoms [δ C 87.15(C-20) and 70.78(C-25)], 8 methyl groups and carbonyl carbon [δ C It had 218.85(C-3)]. 1 Hand 13 As a result of thorough analysis of CNMR data, the aglycone of compound 10 is pseudoginsengenin R1 [(20 S ,24 R )-Damar-3-one-20,24-epoxy-6α,12β,25-triol([(20 S ,24 R It was found that the absolute configuration of C-20 in compound 10 overlapped with the chemical shift (δ) of C-21. C S is inferred from 27.67), and the 24R array is C-24(δ as previously disclosed C It was determined by the chemical shift of 86.09). The two sugar units were determined along with acid hydrolysis data and gas chromatography (GC) analysis results. 1 It was identified as a β-D-glucopyranosyl residue from the H NMR spectrum and the coupling constant of the anomeric proton at the 12-carbon resonance. The glycosidic bond is δ H 6.02(H-1") / δ C 79.49(C-2') and δ H 4.91(H-1') / δC It was determined by the heteronuclear multiple coupling correlation (HMBC) showing a cross peak at 79.79 (C-6), and 2- O -(β-D-glucopyranosyl)-β-D-glucopyranosyl(2- O It was demonstrated that the (β-D-glucopyranosyl)-β-D-glucopyranosyl) residue is connected to C-6 of the aglycone in pseudoginsengenin R1. Each analytical spectrum of the above compound 10 and the key HBMC correlations are shown in Figures 4 to 10.

[0057] As a result of the above analysis, the chemical structure of compound 10 was determined to be (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol, and was named pseudoginsenoside RT8 (PG-RT8).

[0058] PPT among the ginsenosides isolated from the above ginseng seed extract ( P roto P anax T Ginsenosides Rg1 (Compound 1), Rg2 (Compound 2), and Re (Compound 3), which are riol-series ginsenosides, contain three hydroxyl groups in their ginsenoside backbone. PPD ( P roto P anax DGinsenosides of the iol series, such as ginsenoside Rd (Compound 4), ginsenoside Rb1 (Compound 5), and ginsenoside Rb2 (Compound 6), contain two hydroxyl groups in the ginsenoside backbone. On the other hand, Compound 10, a ginsenoside newly isolated and identified in the present invention, has a PPT series backbone, but has a structural difference in that the terminal hydroxyl group of the backbone is a ketone and the linear chain of the ginsenoside is cyclized into a furan ring.

[0059] The molecular formula of compound 10, newly isolated and identified in the present invention, is C 42 H 70 O 15 ,ESI-Q-TOF-MS is m / z is 837.4617 [M+Na] + was, 1 H, 13 The C-NMR spectrum is as shown in the table below.

[0060]

[0061]

[0062] [Test Example 1] Comparison of skin protective efficacy against fine dust 1

[0063] An experiment to confirm the skin protective efficacy against fine dust of the novel ginsenoside GS#10, which is an embodiment of the present invention, was conducted as follows.

[0064] After pre-treating human-derived epithelial cells (Normal Human Dermal Fibroblast (NHDF), ATCC) with the novel ginsenoside GS#10, which is an embodiment of the present invention isolated from the ginseng seed extract, at concentrations of 1, 2, 5, 10, 20, 50, and 100 μg / ml for 2 hours, respectively, 10 μm size (PM10) fine dust (Sigma) was 30 μg / cm² 2The cells were treated at a certain concentration for 24 hours. Then, the cell survival rate was observed by performing an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.

[0065] MTT analysis was performed according to the following method. 100 μl of 10% MTT reagent diluted in serum-free medium was added to a 96-well plate. After incubating at 37°C for 1 hour, the MTT reagent was removed, and 100 μl of formazan was added to each 96-well plate. Then, absorbance was measured at 540 nm, and cell viability was expressed as a percentage (%) of the absolute optical density of each sample relative to the untreated sample.

[0066] As a result, as shown in Fig. 11, the novel ginsenoside GS#10, which is an embodiment of the present invention, was found to inhibit skin cell death caused by fine dust in a concentration-dependent manner, and the effect was prominent starting from 10 μg / ml.

[0067] Next, in order to confirm whether the increase in skin cell viability caused by the novel ginsenoside GS#10, which is an embodiment of the present invention, also appears in other ginseng seed-derived substances, as a comparative example of the present invention, the ginseng seeds used for the isolation of ginsenoside in Example 1 described in detail herein (Seeds of Panax ginsengThe efficacy of the methanol extract (GSE) in inhibiting skin cell apoptosis against fine dust was also analyzed by conducting experiments in the same manner as above. The ginseng seed extract, which is a comparative example of the present invention, contains a small amount of the novel ginsenoside GS#10, which is an embodiment of the present invention, at approximately 0.2 (w / w)%. Figure 12 shows a comparison of the skin cell viability against fine dust of the novel ginsenoside GS#10, which is an embodiment of the present invention, and the ginseng seed extract (GSE). As a result, unlike the novel ginsenoside GS#10, the ginseng seed extract did not exhibit an efficacy in inhibiting skin cell apoptosis from fine dust. This implies that the skin protection efficacy against fine dust is an inherent efficacy of the novel ginsenoside GS#10, which is an embodiment of the present invention.

[0069] [Test Example 2] Comparison of Skin Protection Efficacy Against Fine Dust 2

[0070] The skin protective efficacy against fine dust was compared using the same method as in Test Example 1 above, but the novel ginsenoside GS#10 (isolated from ginseng seed extract), which is an embodiment of the present invention, was compared with three types of ginsenosides (Rg1, Rg3, Rb1; Sigma), which are indicator components of red ginseng, as comparative examples of the present invention. At this time, the concentration of each ginsenoside was 100 μg / ml. The chemical structure of ginsenoside Rg3, which is a comparative example of the present invention, is as follows.

[0071] [Chemical Formula 2]

[0072]

[0073] As a result, as shown in Fig. 13, among the ginsenosides that constitute the indicator components of red ginseng, Rb1 showed almost no skin protective efficacy, and it can be confirmed that the novel ginsenoside GS#10, which is an embodiment of the present invention, has a significantly higher effect of preventing and protecting against skin cell death caused by fine dust than the three types of ginsenosides that constitute the indicator components of red ginseng, by about 2 to 20 times.

[0075] [Test Example 3] Comparison of Respiratory Protective Efficacy Against Fine Dust 1

[0076] An experiment to confirm the respiratory protective efficacy of the novel ginsenoside GS#10, which is an embodiment of the present invention, against fine dust was conducted as follows.

[0077] After pre-treating human lung epithelial cell line A549 (ATCC) with the novel ginsenoside GS#10, an embodiment of the present invention isolated from the ginseng seed extract, at concentrations of 1, 2, 5, 10, 20, 50, and 100 μg / ml for 2 hours, respectively, 10 μm size (PM10) fine dust (Sigma) was 30 μg / cm² 2 The cells were treated at the concentration for 24 hours. Then, the cell survival rate was observed by performing an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay in the same manner as in Test Example 1.

[0078] As a result, as shown in Fig. 14, the novel ginsenoside GS#10, which is an embodiment of the present invention, was found to inhibit lung cell death caused by fine dust in a concentration-dependent manner, and the effect was prominent starting from 5 μg / ml.

[0079] Next, in order to confirm whether the increase in lung cell viability caused by the novel ginsenoside GS#10, which is an embodiment of the present invention, also appears in other ginseng seed-derived substances, as a comparative example of the present invention, the ginseng seeds used for the isolation of ginsenoside in Example 1 described in detail herein Panax ginsengThe efficacy of the methanol extract (GSE) in inhibiting lung cell apoptosis against fine dust was also analyzed by conducting experiments in the same manner as above. The ginseng seed extract, which is a comparative example of the present invention, contains a small amount of the novel ginsenoside GS#10, which is an embodiment of the present invention, at approximately 0.2 (w / w)%. Figure 15 shows a comparison of the lung cell survival rates against fine dust of the novel ginsenoside GS#10, which is an embodiment of the present invention, and the ginseng seed extract (GSE). As a result, unlike the novel ginsenoside GS#10, the ginseng seed extract showed almost no efficacy in inhibiting lung cell apoptosis against fine dust. This implies that the respiratory protective efficacy against fine dust is an inherent efficacy of the novel ginsenoside GS#10, which is an embodiment of the present invention.

[0081] [Test Example 4] Comparison of Respiratory Protective Efficacy Against Fine Dust 2

[0082] The respiratory protective efficacy against fine dust was compared using the same method as in Test Example 3 above, but the novel ginsenoside GS#10 (isolated from ginseng seed extract), which is an embodiment of the present invention, was compared with three types of ginsenosides (Rg1, Rg3, Rb1; Sigma), which are indicator components of red ginseng, as comparative examples of the present invention. At this time, the concentration of each ginsenoside was 100 μg / ml.

[0083] As a result, as shown in Fig. 16, it can be confirmed that the novel ginsenoside GS#10, which is an embodiment of the present invention, has a much superior effect in preventing and protecting against lung cell death caused by fine dust compared to the three types of ginsenosides that constitute the indicator components of red ginseng.

[0085] [Test Example 5] Cytotoxicity

[0086] To exclude the possibility that ginsenosides may affect the skin protective efficacy against fine dust through cytotoxic activity, cell growth in the presence of the novel ginsenoside GS#10, an embodiment of the present invention, was evaluated using CCK (Cell Counting Kit)-8. The experimental method is as follows.

[0087] 10 μl of CCK-8 reagent was added to cultured SH-SY5Y cells (Dojindo, MD, USA) in a 96-well plate, and after being incubated at 37°C for 2 hours, the absorbance was measured at 450 nm. The cell viability was expressed as a percentage (%) of the absolute optical density of each sample relative to the untreated sample. At this time, the concentrations of the novel ginsenoside GS#10, an embodiment of the present invention, included in the culture medium in which the cells were cultured were 0.1, 1, 5, 10, 20, and 50 μM, respectively.

[0088] As a result, as can be seen in Fig. 17, the novel ginsenoside GS#10, an embodiment of the present invention, did not exhibit cytotoxicity up to 50 μM. These results indicate that the novel ginsenoside, an embodiment of the present invention, can exhibit skin and respiratory protective effects against fine dust without having a harmful effect on cell viability.

[0090] These results suggest that the novel ginsenoside PG-RT8, an embodiment of the present invention, possesses skin protective properties against various strong fine dusts and has pharmaceutical potential as a skin protectant and a skin protectant for preventing skin damage from fine dusts.

[0092] A formulation example of a composition according to one embodiment of the present specification is described below, but it can be applied to various other formulations, and this is not intended to limit the present specification but merely to provide a specific description.

[0094] [Formulation Example 1] Softening lotion (skin lotion)

[0095] Softening lotion was prepared using a conventional method according to the composition listed in the table below.

[0096] Formula ingredients Content (weight%) PG-RT8 0.1 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 PEG-12 nonylphenyl ether 0.2 Polysorbate 80 0.4 ethanol 10.0 Triethanolamine 0.1 Preservatives, colorings, fragrances Appropriate amount purified water Remaining amount

[0097] [Formulation Example 2] Nourishing lotion (milk lotion)

[0098] A nourishing lotion was prepared using a conventional method according to the composition listed in the table below.

[0099] Formula ingredients Content (weight%) PG-RT8 0.1 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Beeswax 4.0 Polysorbate 60 1.5 Caprylic / Capric Triglyceride 5.0 Squalan 5.0 Sorbitasesquioleate 1.5 Liquid paraffin 0.5 cetearyl alcohol 1.0 Triethanolamine 0.2 Preservatives, colorings, fragrances Appropriate amount purified water Remaining amount

[0100] [Formulation Example 3] Massage cream

[0101] A massage cream was prepared using a conventional method according to the composition listed in the table below.

[0102] Formula ingredients Content (weight%) PG-RT8 0.1 glycerin 8.0 Butylene glycol 4.0 Liquid paraffin 45.0 beta-glucan 7.0 Carbomer 0.1 Caprylic / Capric Triglyceride 3.0 Beeswax 4.0 Cetearyl glucoside 1.5 Sorbitan sesquioleate 0.9 Vaseline 3.0 paraffin 1.5 Preservatives, colorings, fragrances Appropriate amount purified water Remaining amount

[0103] [Formulation Example 4] Tablet

[0104] After mixing 100 mg of ginsenoside PG-RT8, 400 mg of lactose, 400 mg of corn starch, and 2 mg of magnesium stearate, tablets were manufactured by compressing them according to the usual method of manufacturing tablets.

[0106] [Formulation Example 5] Capsule

[0107] After mixing 100 mg of ginsenoside PG-RT8, 400 mg of lactose, 400 mg of corn starch, and 2 mg of magnesium stearate, a capsule was prepared by filling it into a gelatin capsule according to the standard method of preparing capsules.

[0109] [Formulation Example 6] Granules

[0110] 850 mg of ginsenoside PG-RT, 250 mg of anhydrous crystalline glucose, and 550 mg of starch were mixed, formed into granules using a fluid bed granulator, and then filled into a bag.

[0112] [Formulation Example 7] Drink

[0113] 850 mg of ginsenoside PG-RT, 10 g of glucose, 0.6 g of citric acid, and 25 g of liquid oligosaccharide were mixed, and 300 ml of purified water was added to fill each bottle with 200 ml. After filling the bottles, the mixture was sterilized at 130°C for 4 to 5 seconds to prepare a drink.

[0115] [Formulation Example 8] Caramel formulation

[0116] Caramel was formed by mixing 850 mg of ginsenoside PG-RT, 1.8 g of corn syrup, 0.5 g of skim milk, 0.5 g of soy lecithin, 0.6 g of butter, 0.4 g of hydrogenated vegetable oil, 1.4 g of sugar, 0.58 g of margarine, and 20 mg of salt.

[0118] [Formulation Example 9] Health food

[0119] ingredient Content PG-RT8 100 ㎎ Vitamin mixture Vitamin A acetate 70 ㎍ Vitamin E 1.0 ㎎ Vitamin B1 0.13 ㎎ Vitamin B2 0.15 ㎎ Vitamin B6 0.5 ㎎ Vitamin B12 0.2 ㎍ Vitamin C 10 ㎎ biotin 10 ㎍ Nicotinamide 1.7 ㎎ folic acid 50 ㎍ calcium pantothenate 0.5 ㎎ mineral mixture Ferrous sulfate 1.75 ㎎ zinc oxide 0.82 ㎎ magnesium carbonate 25.3 ㎎ potassium dihydrogen phosphate 15 ㎎ dicalcium phosphate 55 ㎎ potassium citrate 90 ㎎ calcium carbonate 100 ㎎ magnesium chloride 24.8 ㎎

[0120] The composition ratio of the above vitamin and mineral mixture was mixed using ingredients that are relatively suitable for health food as examples, but the mixing ratio may be modified at will. The above ingredients may be mixed according to a conventional method for manufacturing health food, then granules may be prepared and used to manufacture a health food composition according to a conventional method.

[0122] [Formulation Example 10] Health Drink

[0123] ingredient Content PG-RT8 10 ㎎ Citric acid 1000 ㎎ Oligosaccharides 100 g Plum concentrate 2 g Taurine 1 g purified water Remaining amount Total volume 900 ㎖

[0124] As shown in the table above, the remaining amount of purified water is added to make the total volume 900 ml, and the above ingredients are mixed according to a conventional method for manufacturing health drinks. Then, the mixture is stirred and heated at 85°C for about 1 hour, and the resulting solution is filtered, taken into a sterilized 2 L container, sealed and sterilized, and then refrigerated for use in manufacturing a health drink composition.

[0126] [Formulation Example 11] Injectable

[0127] An injectable was prepared by conventional methods according to the composition listed in the table below.

[0128] Formula ingredients Content PG-RT8 10-50 mg Sterile distilled water for injection Appropriate amount pH adjuster Appropriate amount

[0129] The present invention may provide the following embodiments as an example.

[0130] A first embodiment may provide a composition for skin or respiratory protection against fine dust comprising (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient.

[0131] A second embodiment may provide a composition in which, in the first embodiment, the active ingredient has the structure of the following chemical formula 1.

[0132] [Chemical Formula 1]

[0133]

[0134] A third embodiment may provide a composition in which, in the first or second embodiment, the active ingredient is extracted from ginseng seeds.

[0135] The fourth embodiment may provide a composition in which, in any one or more of the first to third embodiments, the active ingredient inhibits damage or death of skin cells caused by fine dust.

[0136] The fifth embodiment may provide a composition in which, in any one or more of the first to fourth embodiments, the active ingredient inhibits damage or death of lung cells caused by fine dust.

[0137] The sixth embodiment may provide a composition comprising, in any one or more of the first to fifth embodiments, the active ingredient in an amount of 0.0001 to 99.9 weight% based on the total weight of the composition.

[0138] The seventh embodiment may provide a composition in which, in any one or more of the first to sixth embodiments, the dosage of the composition is 0.05 mg / kg / day to 10 g / kg / day.

[0139] The eighth embodiment may provide a composition in which, in any one or more of the first to seventh embodiments, the composition is a skin external application composition.

[0140] The ninth embodiment may provide a composition in which, in any one or more of the first to eighth embodiments, the composition is a cosmetic composition.

[0141] The 10th embodiment may provide a composition in which, in one or more of the 1st to 9th embodiments, the composition is a food composition.

[0142] The 11th embodiment may provide a composition in which, in any one or more of the 1st to 10th embodiments, the composition is a pharmaceutical composition.

[0144] The above embodiments are disclosed for the purpose of describing the invention, and such description is not intended to limit the scope of the invention. Accordingly, various modifications, variations, and substitutions may be made by those skilled in the art without departing from the meaning and scope of the invention.

Claims

Claim 1 A food composition for skin or respiratory protection against fine dust comprising (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient. Claim 2 A food composition according to claim 1, wherein the active ingredient has the structure of the following chemical formula 1. [Chemical Formula 1] Claim 3 A food composition according to claim 1, wherein the active ingredient is extracted from ginseng seeds. Claim 4 A food composition according to claim 1, wherein the active ingredient inhibits damage or death of skin cells caused by fine dust. Claim 5 A food composition according to claim 1, wherein the active ingredient inhibits damage or death of lung cells caused by fine dust. Claim 6 A food composition according to claim 1, comprising the active ingredient in an amount of 0.0001 to 99.9 weight% based on the total weight of the composition. Claim 7 A food composition according to claim 1, wherein the dosage of the composition is 0.05 mg / kg / day to 10 g / kg / day. Claim 8 A cosmetic composition for skin protection against fine dust, comprising (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient. Claim 9 A cosmetic composition according to claim 8, comprising the above active ingredient in an amount of 0.0001 to 99.9 weight% based on the total weight of the composition. Claim 10 A pharmaceutical composition for the prevention or treatment of inflammatory skin or respiratory diseases caused by fine dust, comprising (20S,24R)-6-O-β-D-glucopyranosyl(1->2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol, a pharmaceutically acceptable salt thereof, a hydrate thereof, or a solvate thereof as an active ingredient. Claim 11 In claim 10, the above composition is a pharmaceutical composition that is a topical skin preparation composition for preventing or treating inflammatory skin diseases caused by fine dust. Claim 12 A pharmaceutical composition according to claim 10, comprising the above active ingredient in an amount of 0.0001 to 99.9 weight% based on the total weight of the composition. Claim 13 A pharmaceutical composition according to claim 10, wherein the dosage of the above composition is 0.05 mg / kg / day to 10 g / kg / day.

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