KR20120079983A - Novel gallotannin compound isolated from leaf of acer ginnala and antioxidative composition comprising the same as acitive ingredient - Google Patents

Abstract

PURPOSE: A novel gallotannin compound and a composition containing the same for antioxidation are provided to remove DPPH radial and superoxide radical and to enhance antioxidation. CONSTITUTION: A novel gallotannin compound is denoted by chemical formula 1 or 2. The compound is isolated from Acer ginnala. The gallotannin compound is 2,4-di-O-galloyl-1,5-anhydro-D-glucitol and 2,4-di-O-galloyl-1,5-anhydro-D-glucitol. A composition for anti-oxidation contains the gallotannin compound of chemical formula 1 or 2 as an active ingredient. The composition is a cosmetic and functional food composition. The cosmetic composition is manufactured in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, or spray.

Description

Novel Gallotannin Compound Isolated From Leaf of Acer ginnala and Antioxidative Composition Comprising the Same As Acitive Ingredient}

The present invention is a new tree ( Acer ginnala ) relates to a novel gallotannin compound isolated from leaves and an antioxidant composition comprising the same as an active ingredient. More specifically, 2,4-di- O -galloyl-1,5-anhydro-D-glycitol (2,4-di- O- galloyl-1,5-, which is a newly isolated gallotannin compound anhydro-D-glucitol) and 2,4,6-tri- O -galloyl-1,5-anhydro-D-glucitol (2,4,6-tri- O -galloyl-1,5-anhydro -D-glucitol) and the use of these compounds as antioxidants.

Acer ginnala is a member of the family Mapleaceae , which grows widely in the foothills and low mountains. Leaves are used as a source of tannin and diarrhea is used as a medicine in the private sector. It has been reported that the leaves of the new tree have antimicrobial activity against Gram-negative bacteria, such as Shigella flexneri and Pseudomonas aeruginosa , along with antioxidant activity that strongly eliminates DPPH radicals. Studies have reported high levels of gallotannin-based compounds with sugar as the nucleus and gallic acids as substituents. Acertannin, which is known to be contained in a large amount of the leaves of a new tree, is a type of gallotannin, first identified in 1922 (Perkin, AG and Uyeda, YJ Chem. Soc. P66, 1922). The structure was clarified by Bock in 1980. Korean Patent Laid-Open Publication No. 1999-0030624 discloses a Gallicin compound having an antioxidant activity, which is purified and purified from the leaves of a cypress, and a method for separating and purifying the compound, and Korean Patent Laid-Open Publication No. 1999-025781 discloses a separation from a cypress leaf. Purified acertannin-based compounds and their use as natural antioxidants are disclosed.

Free radicals, on the other hand, are chemically highly reactive chemicals, which are caused by various causes in the human body: external factors such as ultraviolet rays, radiation, exhaust gases, tobacco smoke, stress, and metabolism of mitochondria and white blood cells in the body. It is produced through a multi-step chain reaction to cause peroxidation of lipids, which are components of cell membranes. Most of the free radicals are reactive oxygen species (ROS), which include super oxide radicals, hydrogen peroxide (H ₂ O ₂ ), hydroxy radicals, and single-phase oxygen ( Singlet oxygen). The peroxidation of lipids caused by them generates various reactive oxygen species of radicals, peroxides, aldehydes and epoxides and damages DNA, RNA, proteins, cell membranes and cellular structures. In addition, these reactive oxygen species are considered to be the main cause of cancer, tissue damage, aging, skin aging (Black HS, Photochemphotobiol. 46 (2): 213 (1987)). For example, the peroxidation of the membrane lipids that make up the cell may be caused by changes in cell membrane structure and fluidity, increased cellular permeability, leakage of lysosomal enzymes, inactivation of membrane enzymes and transport proteins, generation of covalent crosslinking of lipids and proteins, It causes protein chain breakage, DNA damage, and mutations. In addition, it is known to cause skin aging by peroxidation of skin cell membranes and modification of dermal constituent proteins. Conventionally, many antioxidants such as BHT (2,6-di-tertibutyl-4-methylphenol), BHA (2-tert0butyl-4-methoxyphenol), PG (3,4,5-terhydroxy benzoic acid propyl ester), etc. Was used. However, there are drawbacks such as consumers' rejection of such synthetic antioxidants as the diet and living standards improve. In particular, BHT, a synthetic antioxidant most widely used in the past, has been reported to cause cancer when ingested at 50 mg / kg / day or more.

Numerous papers and patent documents are referenced and cited throughout this specification. The disclosures of the cited papers and patent documents are incorporated herein by reference in their entirety to better understand the state of the art to which the present invention pertains and the content of the present invention.

The present inventors made diligent research efforts to find compounds having antioxidant activity from natural products. As a result, a novel gallotannin compound was isolated from the Acer ginnala leaf extract, and the compound showed strong DPPH radical scavenging activity and superoxide radical scavenging activity, and thus the antioxidant activity was very excellent. The present invention was completed by confirming with.

Accordingly, an object of the present invention is to provide a novel gallotannin compound.

In addition, another object of the present invention to provide an antioxidant composition comprising the novel gallotannin compound as an active ingredient.

The objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

According to one aspect of the present invention, the present invention provides a novel gallotannin compound represented by the following general formula (1) or (2).

According to another aspect of the present invention, the present invention provides a composition for antioxidant comprising a novel gallotannin compound represented by Formula 1 or Formula 2 as an active ingredient.

The present inventors made diligent research efforts to find compounds having antioxidant activity from natural products. As a result, a novel gallotannin compound was isolated from the Acer ginnala leaf extract, and the compound showed strong DPPH radical scavenging activity and superoxide radical scavenging activity, and thus the antioxidant activity was very excellent. The present invention was completed by confirming with.

As used herein, the term “gallotannin” refers to a hydrolyzable tannin compound, in which one or more polyphenol monomers, such as gallic acid, form an ester bond to the hydroxyl group of a polyol carbohydrate such as glucose. Refers to a polymeric compound bound through. The most representative gallotannin compound is pentagalloyl glucose (β-1,2,3,4,6-pentagalloyl- O- D-glucopyranose).

Compound represented by the formula (1) of the present invention is 2,4-di- O -galloyl-1,5-anhydro-D-glucitol (2,4-di- O -galloyl-1,5-anhydro -D-glucitol, and the compound represented by the formula (2) is 2,4,6-tri- O -galloyl-1,5-anhydro-D-glucitol (2,4,6-tri- O -galloyl-1,5-anhydro-D-glucitol), a novel gallotannin compound that has not been reported to date.

In the present invention, the novel gallotannin compound represented by Chemical Formula 1 or 2 is not only purified from natural products, for example, Acer ginnala leaf extract, but also chemically synthesized from the extract. It is apparent to those skilled in the art that it has activity.

According to the conventional method known in the art for extracting an extract from natural products, the nectar leaf extract in the present invention may be separated using a conventional solvent under conditions of conventional temperature and pressure. As an extraction solvent for extracting the leaves of the young tree leaf of the present invention, a solvent which can be generally used in the extraction process may be used. It is preferred to extract using a solvent selected from the group consisting of acetone, ethyl acetate, butyl acetate, dichloromethane, chloroform, hexane and 1,3-butylene glycol.

The gallotannin compound of the present invention can be obtained by a conventional method of separating and purifying a single compound from a natural product extract. For example, filtration using silica gel or celite gel column, size-exclusion chromatography using ion column chromatography, ion-exchange chromatography, partition chromatography, affinity ( affinity chromatography or a combination of these chromatography, but is not limited thereto. Single compounds isolated from natural extracts can be identified through conventional analytical methods known in the art, such as, for example, Matrix-Assisted Laser Desorption Ionization-Time Of Flight Mass Spectroscopy (MALDI-TOF MS), Electron Ionization (EI MS). Mass spectroscopy (CI), chemical ion mass mass spectroscopy (CI), fast atom bombardment ion (FABI), electrospray ion mass spectrometry (ESI-MS), infrared spectroscopy (IR Sectrum), nuclear magnetic resonance (NMR) Magnetic Resonance), CD (Circular Dichroism) and the like, but is not limited thereto.

The novel gallotannin compound of the present invention exhibits strong DPPH radical scavenging activity and superoxide radical scavenging activity, as demonstrated by one specific embodiment of the present invention, which is very excellent in antioxidant activity. Therefore, the novel gallotannin compound may be used as an active ingredient of an antioxidant composition, and in particular, may be used as an active ingredient of a cosmetic composition or a functional food composition having antioxidant activity.

The antioxidant composition of the present invention may be prepared in the form of a cosmetic composition for antioxidant. The cosmetic composition of the present invention may be prepared into any of the formulations conventionally produced in the art, and may be prepared, for example, as a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, But are not limited to, cleansing, oils, powder foundations, emulsion foundations, wax foundations and sprays. More specifically, it may be prepared in the form of a flexible lotion, nutrition lotion, lotion, nutrition cream, massage cream, essence, eye cream, cleansing cream, cleansing foam, cleansing water, pack, spray or powder.

When the formulation of the present invention is a paste, cream, lotion, or gel, the carrier component is animal oil, vegetable oil, wax, paraffin, starch, tracant, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide. This can be used.

In the case where the formulation of the present invention is a powder or a spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component. Especially, in the case of a spray, a mixture of chlorofluorohydrocarbons, propane / Propane or dimethyl ether.

When the formulation of the present invention is a solution or an emulsion, a solvent, a dissolving agent or an emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, , 3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or sorbitan fatty acid esters.

In the case where the formulation of the present invention is a suspension, a carrier such as water, a liquid diluent such as ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, Cellulose, aluminum metahydroxide, bentonite, agar or tracant, etc. may be used.

When the formulation of the present invention is an interfacial active agent-containing cleansing, the carrier component may include aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide Ether sulfates, alkylamidobetaines, aliphatic alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives, or ethoxylated glycerol fatty acid esters.

Ingredients included in the cosmetic composition of the present invention, in addition to the active ingredient and the carrier component, include ingredients conventionally used in cosmetic compositions, for example, such as antioxidants, stabilizers, solubilizers, vitamins, pigments and flavorings Adjuvants may be included.

The antioxidant composition of the present invention may be prepared in the form of a functional food composition for antioxidant. The functional food composition of the present invention includes components that are ordinarily added during the manufacture of food, and includes, for example, proteins, carbohydrates, fats, nutrients, and seasonings. For example, when prepared with a drink, flavoring agents or natural carbohydrates may be included as additional ingredients in addition to the active ingredient. For example, natural carbohydrates include monosaccharides (eg, glucose, fructose, etc.); Disaccharides (eg maltose, sucrose, etc.); oligosaccharide; Polysaccharides (eg, dextrins, cyclodextrins, etc.); And sugar alcohols (eg, xylitol, sorbitol, erythritol, and the like). As the flavoring agent, natural flavoring agents (e.g., taumartin, stevia extract, etc.) and synthetic flavoring agents (e.g., saccharin, aspartame, etc.) can be used.

A summary of the features and advantages of the present invention described above is as follows:

(Iii) The present invention provides a novel gallotanine compound isolated from the leaves of the new tree.

(Ii) Since the novel gallotannin compound of the present invention has a strong antioxidant activity, it can be usefully used as an active ingredient in cosmetics, functional foods and the like which need antioxidant activity.

The present invention provides a novel gallotannin compound 2,4-di-O-galloyl-1,5-anhydro-D-glucitol and 2,4,6-tri-O-galloyl-1,5-an It relates to hydro-D-glucitol. The gallotannin compound of the present invention exhibits strong DPPH radical scavenging activity and superoxide radical scavenging activity and is excellent in antioxidant activity. As a natural antioxidant, it is a raw material for the prevention of adult diseases, functional cosmetics, and health functional foods. Can be used.

1 is a result of measuring the DHHP scavenging activity of the extract of the leaves of the new tree.
2 is a novel gallotannin compound 2,4-di-O-galloyl-1,5-anhydro-D-glucitol and 2,4,6-tri-O-galloyl-1,5 of the present invention. The result of measuring the DHHP scavenging activity of -anhydro-D-glucitol.
Figure 3 is the result of measuring the superoxide scavenging activity of the leaves of the new tree.
4 is a novel gallotannin compound 2,4-di-O-galloyl-1,5-anhydro-D-glucitol and 2,4,6-tri-O-galloyl-1,5 of the present invention. The result of measuring the superoxide scavenging activity of -anhydro-D-glucitol.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for describing the present invention in more detail and that the scope of the present invention is not limited by these embodiments in accordance with the gist of the present invention .

Example

Example  One: God Tree  Isolation of Leaf Extracts and Single Compounds

The leaves of the cypress (2.0 kg) were extracted three times with 80% methanol at room temperature, filtered, and the extract was concentrated under reduced pressure to obtain 674 g of the extract. The extract was made up of six subfractions in a Sephadex LH-20 (2 kg, 10 X 80 cm) resin with mobile phase conditions increasing concentrations from 40% methanol to 10% in methanol by up to 100%. Divided by. Fraction 4 (59.94g) and Fraction 5 (21.73g) were carried out using a water-methanol gradient system on MCI-gel CHP20P (600g, 5 X 60 cm) and ODS (600 g, 5 X 60 cm) resins. As a result, as a single compound, 2,4-di-O-galloyl-1,5-anhydroglucitol and 2,4,6-tri-O-galloyl-1,5-anhydroglusy, respectively Toll was obtained.

Example 2 Structure Identification of Isolated Single Compound

(1) Identification of the structure of 2,4-di-O-galloyl-1,5-anhydroglucitol

Spectroscopic data of the isolated 2,4-di-O-galloyl-1,5-anhydroglucinitol compound of the present invention was as follows.

① ¹ H-NMR (600 MHz, DMSO- d ₆ + D ₂ O); δ 6.97 (4H, each s, 2X galloyl-2H), 4.80 (1H, t, J = 9.6 Hz, H-4), 4.79 (1H, m, H-2), 3.98 (1H, dd, J = 5.4 , 11.1 Hz, H-1a), 3.80 (1H, t, J = 9.3 Hz, H-3), 3.46 (1H, m, H-5), 3.98 (1H, dd, J = 5.4, 11.1 Hz, H -1a), 3.38 (1H, dd, J = 1.8, 11.4 Hz, H-6a), 3.30 (1H, dd, J = 3.9, 11.4 Hz, H-6b), 3.30 (1H, t, J = 11.1 Hz , H-1b).

② ¹³ C-NMR (150 MHz, DMSO- d ₆ + D ₂ O); δ 165.5 ~ 165.8 (C-7 ', 7``), 145.8 (C-3', 3 '', 5 ', 5''), 138.8 ~ 138.9 (C-4', 4 ''), 119.5 ~ 119.7 (C-1 ', 1''), 109.2-109.3 (C-2', 2 '', 6 ', 6''), 79.9 (C-5), 73.0 (C-3), 72.3 (C -2), 72.0 (C-4), 66.5 (C-1), 61.2 (C-6).

③ HRFAB-MS m / z ; 467.0831 [M ⁻ H] ⁻ (calculated for C ₂₀ H ₁₉ O ₁₃ , 467.0826)

Based on the spectroscopic data, it was confirmed that the compound of the present invention is a novel 2,4-di-O-galloyl-1,5-anhydroglucitol compound represented by the following Chemical Formula 1.

[Formula 1]

(2) Structural Identification of 2,4,6-Tri-O-galloyl-1,5-anhydroglucitol

Spectroscopic data of the isolated 2,4,6-tri-O-galloyl-1,5-anhydroglucitol compound of the present invention were as follows.

① ¹ H-NMR (600 MHz, MeOH- d ₄ + D ₂ O); δ 7.09 ~ 7.11 (6H, each s, 3 X galloyl-2H), 5.22 (1H, t, J = 9.6 Hz, H-4), 5.02 (1H, m, H-2), 4.42, (1H, dd , J = 2.4, 12.0 Hz, H-6a), 4.21 (1H, dd, J = 4.8, 12.0 Hz, H-6b), 4.18 (1H, dd, J = 5.4, 11.1 Hz, H-1a), 4.03 (1H, t, J = 9.3 Hz, H-3), 3.85 (1H, m, H-5), 3.45 (1H, t, J = 11.1 Hz, H-1b).

② ¹³ C-NMR (150 MHz, MeOH- d ₄ + D ₂ O); δ 161.1 ~ 161.7 (C-7 ', 7``, 7'''), 145.0 ~ 145.1 (C-3 ', 3'',3''', 5 ', 5'',5''') , 138.5 ~ 138.7 (C-4 ', 4'',4'''), 119.6 ~ 119.8 (C-1 ', 1'',1'''), 108.9 ~ 109.0 (C-2 ', 2'',2''', 6 ', 6'',6'''), 76.8 (C-5), 73.4 (C-3), 71.8 (C-2), 71.1 (C-4), 66.6 ( C-1), 62.7 (C-6).

③ HRFAB-MS m / z; 619.0933 [M ⁻ H] ⁻ (calculated for C ₂₇ H ₂₃ O ₁₇ , 619.0935)

On the basis of the spectroscopic data, it was confirmed that the compound of the present invention is a novel 2,4,6-tri-O-galloyl-1,5-anhydroglucitol compound represented by the following Chemical Formula 2. [Formula 2]

Example 3: Antioxidant Experiment

(One) DPPH Radical Scatters ( radical scavenging activity ) Measure

20 μl of each sample was added to 180 μl of DPPH (200 mM) solution, and the absorbance (OD) was measured at 518 nm after standing for 30 minutes. DPPH radical scavenging ability of each sample was calculated by the formula [1- (sample OD-blank OD) / (control OD-blank OD)] X 100. Vitamin C (Vitamin C) was used as a positive control.

(2) Determination of Superoxide Radical Scavenging Activity

The scavenging ability of the superoxide radical was performed by measuring whether xanthin oxidase reduced the superoxide anion generated in the process of oxidizing xanthin to uric acid. 20 μl of the sample at each concentration was added to 160 μl of the mixture containing phosphate buffer (50 mM), EDTA (0.05 mM), NBT (1 mM), hypoxanthine (0.2 mM), and xanthine. 20 μl of xanthine oxidase (1.2 U / μl) was added thereto, and the absorbance was measured at 590 nm after standing at 37 ° C. for 30 minutes. The superoxide radical scavenging ability of each sample was calculated by the formula [1- (sample OD-blank OD) / (control OD-blank OD)] X 100. Allopurinol was used as a positive control.

(3) Experiment result

① DPPH radical scavenging ability

As a result of measuring the DPPH radical scavenging activity of the extract of the leaves of the young tree, it was confirmed that it has a similar level of DPPH radical scavenging ability compared to the vitamin C which is a positive control group (see FIG. 1). As a result of measuring the DPPH radical scavenging ability of the novel gallotannin compound of the present invention isolated from the leaves of the Cider tree, it was confirmed that the DPPH radical scavenging ability was superior to that of the positive control vitamin C (see FIG. 2).

② Superoxide radical scavenging ability

As a result of measuring the superoxide radical scavenging ability of the extract of the leaves of the tree, it was confirmed that the superoxide radical scavenging ability was comparable to that of the positive control group allopurinol (see FIG. 3). As a result of measuring the superoxide radical scavenging ability of the novel gallotannin compound of the present invention isolated from the leaves of Cider tree, it can be seen that it has a superior superoxide radical scavenging ability at low concentration compared to the positive control allopurinol. (See FIG. 4).

Table 1 is a table showing the results of measuring the IC ₅₀ value of DPPH radical scavenging activity and superoxide radical scavenging ability of the Cinnabar leaf extract.

sample DPPH radical scavenging activity (µg / ml) Superoxide radical scavenging activity (㎍ / ㎖) Cypress Leaf Extract 9.71 ± 1.76 3.19 ± 0.10 Vitamin C 2.51 ± 0.04 - Allopurinol - 1.56 or less

Table 2 is a table showing the results of measuring the IC ₅₀ value of DPPH radical scavenging activity and superoxide radical scavenging activity of the Cinnabar leaf extract.

Samples DPPH radical scavenging activity
(μM) Superoxide radical
Scavenging power (μM) 2,4-di- O -galloyl-1,5-anhydroglucitol 6.87 ± 1.05 2.96 ± 0.14 2,4,6-tri- O -galloyl-1,5-anhydroglucitol 5.72 ± 0.30 2.83 ± 0.09 Vitamin C 13.12 ± 0.33 - Allopurinol - 9.75 ± 1.22

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the same is by way of illustration and example only and is not to be construed as limiting the scope of the present invention. Thus, the substantial scope of the present invention will be defined by the appended claims and equivalents thereof.

Claims (4)

The novel gallotannin compound represented by the following general formula (1) or (2).
[Formula 1]

(2)

An antioxidant composition comprising a novel gallotannin compound represented by the following Formula 1 or Formula 2 as an active ingredient.
[Formula 1]

(2)

The composition of claim 2, wherein the composition is in the form of an antioxidant cosmetic composition.
The composition of claim 2, wherein the composition is in the form of an antioxidant functional food composition.

Images