KR20180013666A - Antioxidant composition comprising uldavioside A - Google Patents

Abstract

The present invention relates to an antioxidant composition comprising an uldavioside A compound isolated from an extract of the dried root skin of Ulmus macrocarpa as an active ingredient. It was confirmed that the uldavioside A compound has excellent antioxidant activity. Accordingly, the uldavioside A compound can be used for developing excellent antioxidant compositions.

Description

Antioxidant composition containing < RTI ID = 0.0 > uldavioside A < / RTI &

The present invention relates to an antioxidative composition containing, as an active ingredient, an uldavioside A compound isolated from a radish extract.

With the development of modern industry, it enjoys rich economic and cultural benefits, but it is accompanied by an awareness of the quality of life due to adult diseases and an aging society, with an increase in life expectancy.

A significant amount of harmful reactive oxygen species (ROS) are produced during the oxidation process for energy production in vivo. These active oxygen species mostly disappear by the in vivo elimination mechanism, but if the balance with the antioxidant defense system is disrupted by instantaneous generation of active oxygen, it causes various diseases (Aruoma O.I., et al., 1998). In addition, as a phenomenon associated with oxidation in vivo, the role of active oxygen derived from oxygen is becoming a cause of aging, and there is a growing interest in elimination thereof (Fridorich I., 1978).

In vivo factors of active oxygen production include cellular metabolism, oxidizing enzyme, bacterial action, and in vitro factors include contaminated air, increased metabolic rate, smoking, carcinogens, certain antibiotics, ultraviolet rays, heat, And excessive intake of

The active oxygen species generally include radicals such as lipid radicals, superoxide radicals and hydroxy radicals (OH), as well as radicals such as hydrogen peroxidase (H ₂ O ₂ ), lipid peroxide, hypochlorous acid, and N-chloramine components. These reactive oxygen species bind to proteins, unsaturated fatty acids, and produce lipid peroxides. They cause damages to DNA, RNA, etc., and damage biological membranes and immune functions, as well as hypertension, arteriosclerosis, heart failure, rheumatoid arthritis, allergies, (Chung HY, 1992; Yagi K., 1987; Lopaczyski W., et al., 2001; Yu BP, 1994; Cooke MS, et al., 1997). Active oxygen is also a major factor in oxidative stress and is closely related to aging, which is a major cause of chronic inflammatory diseases as well as aging-related degenerative diseases. In many animals, the ratio of active oxygen is correlated with the life span of the individual, and the amount of active oxygen is a determinant of the aging process of the individual (Yu BP, 1994).

Retention of homeostasis by intracellular antioxidant activity can restore cellular functions, which can prevent or delay the progression of aging, as well as improve the quality of life by improving chronic inflammation control and immunity. It is known that antioxidants in the process of aging can maintain the activity of defective mitochondria and maintain homeostasis, ultimately enhancing the viability of cells (Lee, YH, et al., 2015; Yang, TK, .

Therefore, research on antioxidants that effectively control these active oxygen species has naturally increased (Heo, Su-Jin, 2005). However, synthetic antioxidants such as dibutyl hydroxy toluene (BHT), butylated hydroxy anisole (BHA), and tert-butylhydroquinone (TBHQ) (Jun BS, et al., 2001). Therefore, development of natural antioxidants having safer and superior efficacy has been continuously tried. Natural antioxidants are mostly antioxidant substances of plant origin, such as omija extract (Jang EJ, et al., 1996), 칡 extract (Han MJ, et al., 1999), citrus flavonoid (Cha JH, et al., 2000) , And mulberry (Kim HJ, et al., 2000).

It is the dried bark of the cork layer of the elm belonging to the Ulmaceae ( Uljiceae ) and the dried skin (Ji Hyunjoon, et al., 1988). In the oriental medicine, it has been reported that it has an excellent effect on germination, anticancer, wound healing and inflammation (Duke JA, 1985) because it is tasteful, nontoxic, swelling, arthritis, gastric ulcer and gastrointestinal disease. There are natural products such as β-sitosterol, phytosterol, stimasterol, tannin, starch, and polysaccharide. (Matsuzaki T., et al., 1985) and the chemical composition (Duke JA et al., 1985) and the separation of friedelin, epifriendelalol and taraxerol, , 1985). However, attempts to study diverse physiological activities of the carrageenan have not been conducted yet.

Accordingly, the inventors of the present invention have completed the present invention by confirming that the uldavioside A compound, which is isolated from the extract of the radish root extract, has an excellent antioxidative activity in the course of studying an antioxidative composition containing the extract of the radish root .

Korean Patent No. 0893166 discloses a proliferation and antioxidative effect of fibroblast of elm extract and Korean Patent Publication No. 2008-0036694 discloses a composition containing a mixed herbal extract However, the compound of the present invention is different from the composition of the present invention because no cryovioside A compound is mentioned.

As a non-patent document, Son B. W., et al. Discloses a cryobioside A compound isolated from elm, but no antioxidative activity of the cryobioside A compound has been described.

Korean Registered Patent No. 0893166, application of a glycoprotein-enriched fraction extracted from elm extract as a cosmetic composition, 2009. 04. 06. Registered. Korean Patent Laid-Open No. 2008-0036694 discloses an antioxidant, antimicrobial, moisturizing, whitening, wrinkle-improving and anti-aging effect for the elderly.

Jae Hyun Lee, et al., Korean Journal of Medicinal Herbs, Korean Medical Index, 295, 1988. ,,,,,,,,,,,, Antioxidative and Cytotoxic Activity of Seaweed by Enzymatic Hydrolysates, Food Industry and Nutrition, 10 (1), 31-41, 2005. Aruoma O.I., Free radical, oxidative stress and antioxidants in human health and disease, J. Am. Oil Chem., Soc., 75, 199-212, 1998. Cha J. H., et al., Effect of citrus flavonoids on the lipid peroxidation contents, Korean J. Postharvest Sci. Technol., 7, 211-217, 2000. Chung H.Y., Aging and carcinogenic mechanisms induced by free radicals, Kor. J. Gerontol., 2, 1-11, 1992. Cooke M. S., et al., Immunogenicity of DNA damage by reactive oxygen species-implications for anti-DNA antibodies in lupus, Free Radic. Biol. Med., 22, 151-159, 1997. Duke J. A., Handbook of medicinal herbs, CRC press, 495, 1985. Fridorich I., The biology of oxygen radicals, Science, 201, 875-881, 1978. Han M. J., et al., Antioxidant effect of ether and ethylacetate fractions of Pueraria thunbergiana extract on perilla oil, Korean J. Soc. Food Sci., 15, 114-120, 1999. Jang E. J., et al., Antioxidant effect of Omija (Shizandra chinensis Baillon) extracts, Korean J. Soc. Food Sci., 12, 372-376, 1996. Jun B. S., et al., Antioxidative activities of fruit extracts of Paulownia tomentosa stued, Korean J. Postharvest Sci. Technol., 8 (2), 231-238, 2001. Kim H.J., et al., Antioxidative activites by water-soluble extracts of Morus alba and Cudrania tricusdata, Korean J. Soc. Agric. Chem. Biotechnol., 43, 148-152, 2000. Lee Y.H., et al., Terrein reduced age-related inflammation induced by oxidative stress through Nrf2 / ERK1 / 2 / HO-1 signaling in aged HDF cells, Cell Biochem. Funct. 33 (7), 479-486, 2015. Lopaczyski W., et al., Antioxidants, programmed cell death, and cancer, Nutr. Res., 21, 295-307, 2001. Son B. W., et al., Catechin glycoside from Ulmus davidiana, Arch., Pharm. Res., 12 (3), 219-222, 1989. Yagi K., Lipid peroxidase and human disease, Chem. Phys. Lipids, 45, 337-341, 1987. Yang T. K., et al., Davallialactone from mushroom reduced premature senescence and inflammation on glucose oxidative stress in human diploid fibroblast cells, J. Agric. Food Chem., 61 (29), 7089-7095, 2013. Yu B. P., Cellular defense against damage from reactive oxygen species, Physiol. Rev., 74, 139-162, 1994.

It is an object of the present invention to provide an antioxidative composition containing an uldavioside A compound, which is isolated from an acanthoi extract, as an active ingredient.

The present invention relates to an antioxidative composition containing, as an active ingredient, a cryoprecipitate A compound represented by the following formula (1).

[Chemical Formula 1]

The antioxidant composition may be a pharmaceutical composition comprising a pharmaceutically acceptable carrier or excipient.

The pharmaceutical composition may be added in an amount of 0.001% by weight to 50% by weight based on the total weight of the composition of the cryovioside A compound.

The pharmaceutical composition may be used for the prevention or treatment of diseases caused by oxides produced by active oxygen.

The diseases caused by the active oxygen-induced oxides include but are not limited to aging, chronic alcoholism, atherosclerosis, cancer, coronary heart disease, cataract, diabetes, Down syndrome, hepatitis, ischemia or reperfusion injury, rheumatoid arthritis, , Renal failure, various degenerative nerve diseases, stroke, atopic dermatitis, bronchitis, epilepsy, chronic obstructive disease, diabetic vascular complication, and myocardial infarction.

In addition, the antioxidant composition may be a cosmetic composition.

The cosmetic composition may be added in an amount of 0.001% by weight to 50% by weight based on the total weight of the composition of the cryobioside A compound.

The cosmetic composition may be used for preventing skin aging.

In addition, the antioxidant composition may be a health functional food having an antioxidative effect.

The health functional food may be added in an amount of 0.001 wt% to 50 wt% based on the total weight of the composition of the urodavioside A compound.

Hereinafter, the present invention will be described in detail.

The urodiboside A compound of Formula 1 may be synthesized according to a conventional method, and may be prepared as a pharmaceutically acceptable salt or may be isolated and purified from the extracts of Leucovorin.

The yukigi is a dried bark and fleshy skin of the elm belonging to the Ulmaceae which has been stripped of the cork layer.

The elm belonging to the elm belongs to Ulmus parvifolia JACQ, Ulmus pumila , Ulmus davidiana var japonica NAK, Ulmus davidiana PLANCH, Ulmus davidiana var suberosa ), Ulmus laciniata , Ulmus macrocarpa HANCE, and Ulmus macrocarpa var, macrophylla NAK, and the like.

The extract can be extracted with water, C1 to C4 lower alcohols or a mixed solution thereof as a solvent. The C1 to C4 alcohols may be selected from the group consisting of methanol, propanol, isopropanol, butanol, and isobutanol. Preferably, it is extracted from water.

The compound isolated from the carrot extract can be purified using column chromatography. The chromatography was carried out by silica gel column chromatography, HP-20 column chromatography, RP-18 column chromatography, LH-20 column chromatography LH-20 column chromatography, and high-performance liquid chromatography.

The diseases caused by the active oxygen-induced oxides include but are not limited to aging, chronic alcoholism, atherosclerosis, cancer, coronary heart disease, cataract, diabetes, Down syndrome, hepatitis, ischemia or reperfusion injury, rheumatoid arthritis, , Renal failure, various degenerative nerve diseases, stroke, atopic dermatitis, bronchitis, epilepsy, chronic obstructive disease, diabetic vascular complication, and myocardial infarction.

The aging may be aging due to oxides produced by active oxygen. Preferably aging promoted by oxides produced by active oxygen.

The degenerative neurological disease may be selected from the group consisting of Alzheimer's disease, Parkinsons ' s disease Huntington's disease, multiple sclerosis, polyneuropathy, epilepsy, ) And dementia.

The antioxidant composition provides a pharmaceutical composition comprising the cryovioside A compound of Formula 1 and an excipient. The compound may be added in an amount of preferably 0.001% by weight to 50% by weight, more preferably 0.001% by weight to 40% by weight, and most preferably 0.001% by weight to 30% by weight, based on the total weight of the composition.

The pharmaceutical compositions may be formulated in the form of powders, granules, tablets, capsules, oral preparations such as suspensions, emulsions, syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method . Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. In the case of formulation, a diluent or excipient such as a filler, an extender, a binder, a wetting agent, a disintegrant, or a surfactant is usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, such as starch, calcium carbonate, Cross, lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppository bases include witepsol, macrogol, tween-61, cacao butter, laurin, glycerogelatin and the like.

The dosage of the pharmaceutical composition of the present invention will depend on the age, sex, body weight of the subject to be treated, the specific disease or condition to be treated, the severity of the disease or condition, the route of administration and the judgment of the prescriber. Dosage determinations based on these factors are within the level of ordinary skill in the art and generally the dosage ranges from 0.01 mg / kg / day to approximately 2000 mg / kg / day. A more preferable dosage is 1 mg / kg / day to 500 mg / kg / day. The administration may be carried out once a day or divided into several times. The dose is not intended to limit the scope of the invention in any way.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, livestock, and humans in various routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection. Since the compound of the present invention has little toxicity and side effects, it can be safely used even for long-term administration for preventive purposes.

In addition, the antioxidant composition may be a cosmetic composition.

The cosmetic composition preferably contains 0.001% by weight to 50% by weight, more preferably 0.001% by weight to 40% by weight, and most preferably 0.001% by weight, of the crybabioside A compound of Formula 1, By weight to 30% by weight.

The cosmetic composition may be in the form of a solution, a gel, a solid or a paste anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, a microcapsule, a microgranule or an ionic form (liposome) Creams, skins, lotions, powders, ointments, sprays or conical sticks. It can also be prepared in the form of a foam or an aerosol composition further containing a compressed propellant.

The cosmetic composition may be added to the cryoprecipitate A compound of the present invention in the form of a lipid, an organic solvent, a solubilizer, a thickening agent and a gelling agent, a softening agent, a suspending agent, a stabilizer, a foaming agent, a fragrance, Or any other commonly used in cosmetics such as nonionic emulsifiers, fillers, sequestering and chelating agents, preservatives, vitamins, barrier agents, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or cosmetics ≪ RTI ID = 0.0 > cosmetics. ≪ / RTI >

The cosmetic composition may be selected from lotion, gel, oil-soluble liquid, cream, ointment and essence.

The cosmetic composition may be used for preventing skin aging.

In addition, the antioxidant composition may be a health functional food having an antioxidative effect.

The health functional food provides a health functional food having an antioxidative effect including a crodobiose A compound of formula (1) and a pharmaceutically acceptable food-aid additive.

The above-mentioned urodibioside A compound is preferably added in an amount of 0.001 to 50% by weight, more preferably 0.001 to 30% by weight, and most preferably 0.001 to 10% by weight based on the total weight of the total food .

The health functional food includes forms such as tablets, capsules, pills, and liquids. Examples of the foods to which the extract of the present invention can be added include various foods, beverages, gums, tea, Functional foods.

The present invention relates to an antioxidative composition containing, as an active ingredient, an uldavioside A compound isolated from a carrot extract, and it has been confirmed that the above-mentioned urodiboside A compound has an excellent antioxidative activity.

Accordingly, the present invention can be used for the development of a composition for preventing or treating various diseases caused by oxides produced by active oxygen, an anti-aging-related cosmetic composition, and a health functional food for antioxidation It is expected.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a diagram showing the correlation between ¹ H- ¹ H COZY (bold lines) and ¹ H- ¹³ C HMBC for an uldavioside A compound of the present invention (arrow).
Figure 2 shows the C2C12 (A) and NIH3T3 (B) hydrogen peroxide (H ₂ O ₂₎ generated from the inhibitory activity of the cells of the invention A seed weep rainy compound.
Figure 3 shows the DPPH radical scavenging activity of the cryovioside A compound of the present invention.
Figure 4 shows the total antioxidant activity of the cryovioside A compound of the present invention.
FIG. 5 shows the cytotoxicity results of C2C12 (A) and NIH3T3 (B) cells of the cryovioside A compound of the present invention.

Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.

≪ Example 1: Isolation of cryptobioside A compound >

Udon thunbergii was purchased from Ulmus davidiana Planch dried at the farm. After adding 1 L of purified water to 10 g dried rhizome, the mixture was treated with a semi-continuous low-temperature vacuum extractor at 25 ° C for 12 hours. The Rhizopus japonicus extract was extracted three times, and impurities were removed using a filter paper to obtain hot water extract . The diaion HP-20 column chromatography (Diaion HP-20 column chromatography) was performed on the isocratic elution conditions with 30% methanol, 50% methanol, 70% methanol and 100% Fractions were obtained (Fr. E-1 to E-3). Of these, Fr. E-1 was subjected to sephadex LH-20 column chromatography according to the elution conditions of 70% methanol and the like to separate the urodibioside A compound (9 mg).

Example 2: Identification of the physico-chemical structure of the urodavioside A compound [

Uldaviosidea;

Light brown powder;

≪ ¹ > H NMR and < ¹³ > C NMR data are shown in Table 1 below;

ESI-MS [M + H] ⁺ m / z 423.1, C ₂₀ H ₂₂ O ₁₀ .

Position Cuda Bioside A [delta] _C (ppm) ^a [delta] _H (ppm) ^b 2 82.9 4.59 (1H, d, J = 6.8 Hz) 3 68.6 3.98 (1 H, m) 4 28.4 2.84 (1H, dd, J = 16.5, 5.5 Hz)
2.53 (1H, dd, J = 16.5,8.3 Hz)  4a 103.2 5 158.2 6 97.2 6.12 (1H, d, J = 2.0 Hz) 7 157.6 8 96.8 6.06 (1H, d, J = 2.0 Hz)  8a 156.9 9 132.1 10 115.2 6.82 (1H, d, J = 2.0 Hz) 11 146.2 12 146.3 13 116.1 6.75 (1H, dd, J = 8.2 Hz) 14 119.9 6.70 (1H, dd, J = 8.2, 2.0 Hz)   One' 108.7 5.47 (1H, d, J = 3.4 Hz)   2' 78.2 4.13 (1H, d, J = 2.7 Hz)   3 ' 80.3   4' 75.4 4.07 (1H, d, J = 9.6 Hz)
3.84 (1H, d, J = 9.6 Hz)   5 ' 64.9 3.60 (2H, m) ^{a 13 C NMR (150 MHz in} CD 3 OD) spectroscopy data
^{b 1} H NMR (600 MHz in CD ₃ OD) spectroscopy data

< Example  3. Culture of animal cells>

In order to confirm the activity of the cryovioside A compound of the present invention, animal cells were cultured. At this time, animal cells were used as myocyte host C2C12, fibroblast host NIH3T3 cells. Each of the cells was inoculated into DMEM (Dulbecco's Modified Eagle's Medium) medium containing 10% fetal bovine serum (FBS), 100 U / ml of penicillin and 100 μg / ml of streptomycin, % CO ₂ incubator.

Example 4. Confirmation of antioxidative effect of the compound of cryptobiose A &gt;

Example 4-1. Confirmation of hydrogen peroxide generation inhibition of urodibioside A compound

A hydrogen peroxide (H ₂ O ₂ ) detection kit was used to confirm the antioxidative effect of the cryoprecipitate A compound of the present invention.

The C2C12 and NIH3T3 cells cultured in Example 3 were divided into 24 × 10 ⁵ cells per well and cultured for 24 hours. Cells were seeded with 1 μM, 10 μM and 20 μM of the urodibioside A compound The cells were pre-reacted for 1 hour and then treated with glucose oxidase so as to produce hydrogen peroxide at a concentration of 7 mU / ml and cultured for 24 hours. At this time, C2C12 or NIH3T3 cells, which were not treated with either, were used as a control.

H ₂ O ₂ detection kit (biovision, USA) was used to measure the hydrogen peroxide secreted in the cell culture medium. The experimental method was carried out based on the experimental method provided in the kit, and the results are shown in FIG.

As shown in FIG. 2, it was found that the amount of hydrogen peroxide decreased in both of the C2C12 (3A) and NIH3T3 (3B) cells depending on the treatment concentration of the cryovioside A compound of the present invention.

Example 4-2. Identification of free radical scavenging ability of urodibioside A compound

DPPH (?,? -Diphenyl-? -Picrylhydrazyl) radical scavenging ability was confirmed in order to confirm the antioxidative effect according to the treatment of the urobiosid A compound of the present invention. The DPPH radical scavenging ability is based on the principle that the antioxidant in the sample reacts with the DPPH reagent, which is a free radical, to decolorize from purple to yellow by eliminating free radicals.

0.2 ml of the compound of the present invention was mixed with 0.8 ml of a 0.4 mM DPPH solution dissolved in ethanol, followed by reaction in a dark room for 10 minutes. At this time, the urodibioside A compound was treated to give final concentrations of 1, 10, 20 and 50 uM, and ascorbic acid and butylated hydroxyanisol (BHA) were treated at the same concentration as a control group. After the reaction was completed, the absorbance was measured at 517 nm, and the DPPH radical scavenging activity was calculated using the following equation. The results are shown in FIG. 3 and Table 2.

DPPH radical scavenging activity (%) = (1-sample group absorbance - blank absorbance) × 100

Configuration Free radical scavenging ability
(IC ₅₀ , uM) Cuda Bioside A 8.6 ± 0.4 Ascorbic acid 8.4 ± 0.8 BHA 19.9 ± 0.3

As shown in FIG. 3, the DPPH radical scavenging ability of the urobiosid A compound of the present invention was significantly higher than that of ascorbic acid used as a control at a concentration of 1 uM, and was similar to that of ascorbic acid at a concentration of 10 uM or more. On the other hand, the DPPH radical scavenging ability of the urobiosid A compound of the present invention was significantly higher at all concentrations than the other control group, butylhydroxyanisole (BHA).

In addition, as shown in Table 2, it was confirmed that the IC ₅₀ value against the DPPH radical scavenging ability of the culdoubioside A compound of the present invention was similar to that of ascorbic acid used as the control group and lower than that of butylhydroxyanisole (BHA) .

As a result, it was found that the free radical scavenging activity of the compound of the present invention was very good, similar to that of ascorbic acid.

Example  4-3. Cryobio seed  A total of compounds Antioxidant ability (total antioxidant capacity)

The total antioxidant capacity of the cryoprecipitate A compound of the present invention was measured using a total antioxidant capacity colorimetric kit of Biovision Co. (measuring the conversion of Cu ²⁺ to Cu ⁺ by antioxidant activity) Were performed according to the experimental method provided by Biovision.

Trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carbocylic acid) used as a standard curve was diluted by concentration and ascorbic acid used as a cobaltoside A compound or a control group and butylhydroxy After preparing the sole for 10 μM, 10 μl of Cu ²⁺ was added to each sample, reacted at room temperature for 90 minutes, and the absorbance value was measured at 570 nm.

A standard curve was obtained by using the absorbance value of each concentration of Trolox, and the total antioxidative capacity of each sample was calculated through the obtained standard curve. The results are shown in FIG.

As shown in FIG. 4, the total antioxidative capacity of the compound of the present invention was significantly higher than that of butylhydroxyanisole used as a control, and was similar to that of ascorbic acid.

From these results, it can be seen that the compound of the present invention has a remarkably superior antioxidative activity.

Example 5: Cytotoxicity of cryovioside A compound < RTI ID = 0.0 >

The cytotoxicity of the cryovioside A compound of the present invention was confirmed by the MTT assay method.

The C2C12 and NIH3T3 cells cultured in the above Example 3 were seeded in a 48 well plate at 1 x 10 &lt; ⁵ &gt; cells Cells were cultured for 24 hours, and then cultured in DMEM (Dulbecco's Modified Eagle's Medium) containing 0.5% FBS (fetal bovine serum), 100 U / ml penicillin and 100 μg / ml streptomycin ) Medium and incubated for 18 hours. After 18 hours of incubation, the cryovioside A compound was treated with 1 [mu] M and 10 [mu] M and cultured for 24 hours. At this time, C2C12 and NIH3T3 cells, which were not treated with anything, were used as a control. After 24 hours, the cells were treated with MTT solution at a concentration of 100 μg / ml and reacted at 37 ° C. for 3 hours. The culture solution was removed, and the formazan precipitate formed was dissolved in 200 μl of DMSO (dimethyl sulfoxide) And the results are shown in Fig.

As shown in Fig. 5, the survival rates of C2C12 (3A) and NIH3T3 (3B) cells treated with the cucurbitidoside A compound were increased compared to the control. As a result, it was found that the cryovioside A compound of the present invention did not show cytotoxicity.

&Lt; Formulation Example 1 >

Formulation Example 1-1. Manufacture of tablets

200 g of the compound of the present invention was mixed with 175.9 g of lactose, 180 g of potato starch and 32 g of colloidal silicic acid. A 10% gelatin solution was added to the mixture, followed by pulverization and passed through a 14-mesh sieve. This was dried, and a mixture obtained by adding 160 g of potato starch, 50 g of activity and 5 g of magnesium stearate was made into tablets.

Formulation Example 1-2. Injection preparation

1 g of the compound of the present invention, 0.6 g of sodium chloride and 0.1 g of ascorbic acid were dissolved in distilled water to make 100 ml. This solution was placed in a bottle and sterilized by heating at 20 DEG C for 30 minutes.

&Lt; Preparation Example 2: Preparation of cosmetic composition >

Formulation Example 2-1. Cream manufacturing

4 g of sitosterol, 3 g of polyglyceryl 2-olate, 0.7 g of ceramide, 2 g of cetearate-4, 3 g of cholesterol, 0.4 g of dicetyl phosphate, 5.0 g of concentrated glycerin, 22 g of Spluaru oil 0.5 g of carboxyvinyl polymer, 0.5 g of triethanolamine, a small amount of preservative and perfume, and purified water were mixed to a total weight of 100 g, and a nutritional cream was prepared according to a conventional cream preparation method.

Formulation Example 2-2. Lotion manufacturing

0.05 g of polypyrrolidone, 0.1 g of oleyl alcohol, 0.2 g of polyoxyethylene monooleate, 0.2 g of perfume, 0.1 g of p-hydroxybenzoic acid methyl ester, a small amount of antioxidant and a small amount of dye were mixed and dissolved in 8 g of 95% ethanol. 0.05 g of the urodibioside A compound of the present invention and 5 g of glycerin were dissolved in 85.33 g of purified water. The mixture was added to the mixture and stirred to prepare a lotion containing the extract of the mixed herb.

Preparation Example 2-3. Emulsion manufacturing

1.2 g of cetyl alcohol, 10 g of squalane, 2 g of vaseline, 0.2 g of p-hydroxybenzoic acid ethyl ester, 1 g of glycerin monoestearate, 1 g of polyoxyethylene (20 mols of addition) monooleate and 0.1 g of perfume were heated and mixed at 70 캜 0.5 g of the compound of the present invention, 5 g of dipropylene glycol, 2 g of polyethylene glycol-1,500, 0.2 g of triethanolamine and 76.2 g of purified water were dissolved by heating at 75 占 폚. The mixture was emulsified by mixing and then cooled to prepare an oil-in-water (O / W) type emulsion.

Formulation Example 2-4. Serum preparation

To 5 g of 95% ethyl alcohol, 1.2 g of polyoxyethylene sorbitan monooleate, 0.3 g of quitrose, 0.2 g of sodium hyaluronate, 0.2 g of vitamin E-acetate, 0.2 g of sodium permanganate, 0.1 g of paraoxybenzoic acid ethyl ester, 1 g of the compound of the present invention was mixed with an appropriate amount of coloring matter to prepare a serum.

&Lt; Formulation example 3. Food production >

Formulation Example 3-1. Manufacture of cooking seasonings

The cryobioside A compound of the present invention was added to the cooking seasoning at 1 wt% to prepare a cooking sauce for health promotion.

Formulation Example 3-2. Manufacture of flour food products

A food for health promotion was prepared by adding 0.1% by weight of the compound of the present invention to wheat flour, and preparing bread, cake, cookie, cracker and noodle by using this mixture.

Formulation Example 3-3. Manufacture of soups and gravies

The health-enhancing soup and juice were prepared by adding 0.1% by weight of the compound of the present invention to the soup and the juice.

Formulation Example 3-4. Manufacture of dairy products

The dulabioside A compound of the present invention was added to milk in an amount of 0.1% by weight, and various dairy products such as butter and ice cream were prepared using the milk.

Formulation Example 3-5. Manufacture of vegetable juices

0.5 g of the compound of the present invention was added to 1,000 ml of tomato juice or carrot juice to prepare vegetable juice for health promotion.

Formulation Example 3-6. Manufacture of fruit juices

0.1 g of the compound of the present invention was added to 1,000 ml of apple juice or grape juice to prepare vegetable juice for health promotion.

Claims (10)

1. An antioxidative composition comprising, as an active ingredient, a cryoprecipitate A compound represented by the following formula (1).
[Chemical Formula 1]

3. The method of claim 2,
Wherein said composition is a pharmaceutical composition comprising a pharmaceutically acceptable carrier or excipient. 3. The method of claim 2,
Wherein said pharmaceutical composition is added in an amount of from 0.001% to 50% by weight, based on the total weight of the composition, of the urodiboside A compound. The method according to claim 2 or 3,
Wherein said pharmaceutical composition is used for the prevention or treatment of diseases caused by oxides produced by active oxygen. 5. The method of claim 4,
The diseases caused by the active oxygen-induced oxides include but are not limited to aging, chronic alcoholism, atherosclerosis, cancer, coronary heart disease, cataract, diabetes, Down syndrome, hepatitis, ischemia or reperfusion injury, rheumatoid arthritis, Wherein the antioxidant composition is at least one disease selected from the group consisting of renal failure, various degenerative nerve diseases, stroke, atopic dermatitis, bronchitis, epilepsy, chronic obstructive disease, diabetic vascular complication, and myocardial infarction. The method according to claim 1,
Wherein the composition is a cosmetic composition. The method according to claim 6,
Wherein the cosmetic composition is added in an amount of 0.001% to 50% by weight based on the total weight of the composition of the cryovioside A compound. 8. The method according to claim 6 or 7,
Wherein the cosmetic composition is for preventing skin aging. The method according to claim 1,
Wherein the composition is a health functional food having an antioxidative effect. 10. The method of claim 9,
Wherein said health functional food is added in an amount of 0.001 wt% to 50 wt% based on the total weight of the composition of the urodavioside A compound.

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