KR20230027384A - An antioxidant composition comprising, as an active ingredient, an enzyme-treated product of sargassum siliquastrum or a polysaccharide isolated therefrom - Google Patents

Abstract

The present invention relates to a composition comprising an enzyme-treated product of Sargassum siliquastrum, and a polysaccharide isolated therefrom as an active component. By using the composition containing the enzyme-treated product of Sargassum siliquastrum or the polysaccharide isolated therefrom as the active component, it is possible to effectively alleviate oxidative stress.

Description

An antioxidant composition comprising, as an active ingredient, an enzyme-treated product of sargassum siliquastrum or a polysaccharide isolated therefrom}

It relates to an anti-oxidation composition comprising an enzymatically treated product of a pretzel cap or a polysaccharide isolated therefrom.

As we enter an aging society in which the average life span of humans has increased due to the improvement of living standards and the remarkable development of medical technology in the 21st century, the desire for healthy aging is getting stronger. In response to these times, the market for health functional foods is growing rapidly. Oxidative stress refers to a state in which the level of oxidation in the body, which was maintained almost constant by the balance of the reactive oxygen production system and the scavenging system, is out of balance due to various factors such as drugs, radiation, ischemia, and aging. , excessive free radicals are generated to break the balance of the body's antioxidant mechanism, and as a result of oxidative stress, cell damage (especially cell membrane lipids and oxidation), mutation by DNA base modification, and induction of cell death (apotosis) cause arteriosclerosis, It is the cause of various diseases such as degenerative brain disease, stroke, Parkinson's disease, Huntington's disease, diabetes, cancer, Alzheimer's, dementia, and aging. The cause of oxidative stress is reactive oxygen species (ROS), which are free radicals. In order to remove ROS in vivo, intake of foods containing enzymes such as SOD (superoxide dismutase) or foods containing β-carotene, vitamins C and E, and phenolic compounds has been made, but development of more effective alternatives is still required. It is coming. Recently, studies on active ingredients derived from natural products have been conducted to improve oxidative stress that causes various diseases. Compositions useful for the prevention, improvement, and treatment of oxidative stress-related diseases such as arteriosclerosis, dementia, and cancer using active ingredients have been disclosed, but still isolation and development of novel active ingredients having oxidative stress improving activity in various natural products This is being desired.

Sargassum siliquastrum is a seaweed belonging to the brown algae family, Sargassum siliquastrum, and is mainly distributed in Korea and Japan. It mainly lives on rocks from the low tide to the midday, and usually grows to 2-3m or several tens of meters in length. It has various shapes depending on the season and topography, but generally the leaves at the base are wide, do not have double sawtooth, and the branches are three-ringed and twisted in a twisted shape. Air bubbles are attached to the upper part, so it extends straight up even in the sea.

It has been reported that capsicum chromanol purified from capsular caps has an antioxidant effect (Korean Patent No. 10-0765160), but the antioxidant effect of extracts obtained by treating capsicum capsicum with enzymes and polysaccharides isolated therefrom has been reported. never happened

Republic of Korea Patent No. 10-0765160

One embodiment provides an anti-oxidation composition comprising, as an active ingredient, an enzymatically-treated product of tangle cap or a polysaccharide isolated therefrom.

Another embodiment provides a pharmaceutical composition for the prevention or treatment of peripheral vascular disease, allergic dermatitis, skin cancer, or dermatosis, containing as an active ingredient an enzymatically treated product of capsular capitulum or a polysaccharide isolated therefrom.

Another embodiment provides a cosmetic composition comprising an enzymatically treated product of capsular capella or a polysaccharide isolated therefrom.

Another embodiment provides a food composition comprising an enzymatically treated product of capsular caps or a polysaccharide isolated therefrom.

Another embodiment provides a method for producing a polysaccharide derived from capsular capsicum, comprising the step of obtaining an extract from capsicum capsicum by treating capsicum capsicum with an enzyme.

One aspect provides an anti-oxidation composition comprising an enzymatically treated product of a pretzel cap or a polysaccharide isolated therefrom.

As used herein, the term “enzyme-treated product of capsular caps” refers to a material obtained by reacting capsicums with capsular caps and enzymes.

As used herein, the term "antioxidation" refers to preventing oxidative stress by stabilizing active oxygen by donating electrons to active oxygen. Human skin itself has an antioxidant network that responds to active oxygen. However, when it is difficult for the endogenous antioxidant network to handle oxidative stress occurring inside and outside the body, active oxygen is not sufficiently removed and causes tissue damage. Therefore, when oxidative stress increases due to external factors such as ultraviolet rays, smoking, and environmental pollutants, aging progresses such as pigmentation and wrinkles in the skin, and the risk of skin cancer increases. Moreover, the inherent antioxidant capacity of the human body gradually decreases with age and is not sufficient to effectively cope with oxidative stress. Antioxidants supplement the insufficient antioxidant network.

According to one embodiment, the enzyme may be a glycolytic enzyme.

The glycolytic enzyme may be one or more selected from the group consisting of amylo glucosidase (AMG), celluclast, termamyl, ultraflo, and viscozyme. , but not limited thereto.

According to one embodiment, the "enzyme" may be a cellulase.

The "cellulase" may be, for example, "Celluclast 1.5L FG", but is not limited thereto. The "Celluclast 1.5L FG" is 1,4-(1,3:1,4)-β-D-Glucan 4-glucano-hydrolase extracted from Trichoderma reesei ATCC 26921, Enzyme Commission (EC) Number 3.2.1.4 , a product of Novozyme (Bagsvaerd, Denmark), and its enzyme activity is 700 Endoglucanase unit/g.

Another aspect provides a pharmaceutical composition for preventing or treating peripheral vascular disease, allergic dermatitis, or skin cancer, comprising an antioxidant composition containing, as an active ingredient, an enzymatically treated product of capsicum capsula or a polysaccharide isolated therefrom as an active ingredient.

As used herein, the term "active ingredient" means a component that exhibits the desired activity alone or that can exhibit activity in combination with a carrier that is not active itself.

As used herein, the term "prevention" refers to any action that suppresses or delays the onset of atopic dermatitis by administration of the pharmaceutical composition according to the present invention.

As used herein, the term "treatment" refers to all activities in which symptoms of atopic dermatitis are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.

For administration, the pharmaceutical composition may be preferably formulated as a pharmaceutical composition by including at least one pharmaceutically acceptable carrier in addition to the above-described enzymatically treated product of the capsular capella or the polysaccharide isolated therefrom.

Formulations of the pharmaceutical composition may be granules, powders, tablets, coated tablets, capsules, suppositories, solutions, syrups, juices, suspensions, emulsions, drops, or injectable solutions. For example, for formulation in the form of tablets or capsules, the active ingredient may be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. In addition, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included in the mixture. Suitable binders include, but are not limited to, starch, gelatin, natural sugars such as glucose or beta-lactose, corn sweeteners, natural and synthetic gums such as acacia, tracacanth or sodium oleate, sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum, and the like.

In compositions formulated as liquid solutions, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and One or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare formulations for injections such as aqueous solutions, suspensions, and emulsions, pills, capsules, granules, or tablets.

The pharmaceutical composition of the present invention can be administered orally or parenterally, and in the case of parenteral administration, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, transdermal administration, etc. can be administered.

A suitable dosage of the pharmaceutical composition of the present invention may be determined by factors such as formulation method, administration method, patient's age, weight, sex, morbid condition, food, administration time, administration route, excretion rate and reaction sensitivity. .

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulation using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily performed by those skilled in the art. or it may be prepared by incorporating into a multi-dose container.

According to one embodiment, the pharmaceutical composition may be an external preparation for skin.

External preparations can be applied in the form of patches, bands, emulsions, ointments, packs, gels, creams, lotions, liquids or powders, and as cosmetics, softening lotion, nutrient lotion, massage cream, nutrient cream, moisturizing cream, functional lotion, mist , It can be applied as a pack, gel or skin adhesive type formulation. Therefore, in order to be used as the external preparation, components commonly used in external preparations such as cosmetics or pharmaceuticals, such as aqueous components, oily components, powder components, alcohols, moisturizers, thickeners, ultraviolet absorbers, whitening agents, preservatives, antioxidants, surfactants, and fragrances , Colorants, various skin nutrients, etc. may be appropriately incorporated into the composition as needed. The external preparations include metal sequestering agents such as disodium edetate, trisodium edetate, sodium citrate, sodium polyphosphate, sodium metaphosphate, and gluconic acid, caffeine, tannin, bellapamil, licorice extract, glabridin, and calin fruit hot water extract, various herbal medicines, tocopherol acetate, glycylitnic acid, tranexamic acid and its derivatives or salts, vitamin C, magnesium ascorbate, ascorbic acid glucoside, arbutin, kojic acid, glucose, fructose, trehalose Sugars such as can also be blended appropriately.

Another aspect provides a cosmetic composition comprising an enzymatically treated product of capsular caps or a polysaccharide isolated therefrom.

Products to which the cosmetic composition of the present invention can be added include, for example, various skin lotions, skin softeners, skin toners, astringents, lotions, milk lotions, nutrient lotions, massage creams, nutrient creams, hand creams, foundations, and makeup. It may be a base, twin cake, mascara, essence, nutritional essence, pack, soap, cleansing foam, cleansing lotion, cleansing cream, body lotion, or body cleanser.

When the cosmetic composition of the present invention is prepared in the above-described formulation, it may contain various bases and additives necessary and suitable for formulating the formulation. In addition, the types and amounts of these components can be easily selected by those skilled in the art.

For example, when the formulation of the present invention is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tracanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide as a carrier component etc. can be used.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizing agent or emulsifying agent is used as a carrier component, such as water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1 ,3-butyl glycol oil, glycerol fatty esters, polyethylene glycol or fatty acid esters of sorbitan may be used.

When the formulation of the present invention is a suspension, as a carrier component, a liquid diluent such as water, ethanol or propylene glycol, an ethoxylated isostearyl alcohol, a suspending agent such as polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystals Star cellulose, aluminum metahydroxide, bentonite, agar or tracanth and the like may be used.

When the formulation of the present invention is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and in particular, in the case of a spray, additional chlorofluorohydrocarbon, propane / May contain a propellant such as butane or dimethyl ether.

When the formulation of the present invention is surfactant-containing cleansing, as carrier components, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether Sulfates, alkylamidobetaines, fatty alcohols, fatty acid glycerides, fatty acid diethanolamides, vegetable oils, lanolin derivatives or ethoxylated glycerol fatty acid esters and the like can be used.

In addition, the cosmetic composition of the present invention can be used alone or overlapping, or overlapping with other cosmetic compositions other than the present invention. In addition, the cosmetic composition according to the present invention can be used according to a conventional method of use, and the number of times of use can be varied according to the user's skin condition or taste.

Another aspect provides a food composition comprising an enzymatically treated product of a pretzel cap or a polysaccharide isolated therefrom.

The food composition according to the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, confectionery, diet bars, dairy products, meat, chocolate, pizza, ramen, other noodles, chewing gum, ice cream, vitamin complexes, and health supplements. .

The food composition of the present invention may include ingredients commonly added during food production, in addition to containing the enzymatically treated product of the pretzel cap or the polysaccharide isolated therefrom, for example, proteins, carbohydrates, fats, nutrients, seasoning and flavoring agents. Examples of the aforementioned carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides such as conventional sugars such as dextrins and cyclodextrins and sugar alcohols such as xylitol, sorbitol and erythritol. As flavoring agents, natural flavoring agents [thaumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)] and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is made into drinks and beverages, citric acid, high fructose corn syrup, sugar, glucose, acetic acid, malic acid, fruit juice, and Various plant extracts and the like may be further included.

Another aspect provides a method for producing a polysaccharide derived from capsular capsicum, comprising the step of obtaining an extract from capsicum capsicum by treating the capsicum capsicum with an enzyme.

The celluclast enzyme is treated and reacted with the cap half of the pretzel to obtain an extract, and after the reaction, the obtained extract is centrifuged to filter and inactivate to prepare a final celluclast extract.

The centrifugation may be performed at 3000 to 5000 rpm, preferably 4000 rpm for 10 to 30 minutes, preferably 20 minutes.

The filtering and inactivation may be performed at 80 to 125 °C, 90 to 115 °C, preferably 95 to 100 °C, within 20 minutes, preferably for 5 to 10 minutes.

The prepared celluclast extract may be treated with 80% or more ethanol, preferably 100% ethanol, and reacted at 2 to 6 ° C, preferably 4 ° C for 10 to 14 hours, preferably 12 hours.

After the reaction, the primary polysaccharide (SSS1) can be obtained by centrifugation (4000 rpm, 20 minutes), and immediately the supernatant is treated with 80% or more ethanol, preferably 100% ethanol, in the same manner as above. Primary (SSS2), tertiary (SSS3), and quaternary (SSS4) polysaccharides can be obtained.

The polysaccharide of the tangle cap enzyme extract is from the group consisting of fucose, rhamnose, galactose, glucose, mannose, xylose and arabinose It may consist of one or more selected monosaccharides, and may have a total molecular weight of 20,000 to 400,000 Da.

Specifically, the total molecular weight of the polysaccharide may be 20,000 Da or more, 40,000 Da or more, 60,000 Da or more, 80,000 Da or more, 100,000 Da or more, 150,000 Da or more, 200,000 Da or more, 250,000 Da or more, 300,000 Da or more, or 350,000 Da or more. , In addition, it may be 400,000 Da or less, 350,000 Da or less, 300,000 Da or less, 250,000 Da or less, 200,000 Da or less, 150,000 Da or less, 100,000 Da or less, 80,000 Da or less, 60,000 Da or less, or 40,000 Da or less.

According to one embodiment, the step of obtaining the extract may be to treat the enzyme for 22 hours to 26 hours at 40 to 60 ℃, pH is 4 to 5.

The temperature of the step of obtaining the extract may be treated at 20 to 80 °C, 30 to 70 °C, 40 to 60 °C, preferably 50 °C.

The pH of the step of obtaining the extract may be treated at pH 3.6 to 5.4, pH 3.8 to 5.2, pH 4 to 5, preferably pH 4.5.

Obtaining the extract may be performed for 20 to 28 hours, 22 to 26 hours, preferably 24 hours.

According to one embodiment, the enzyme may be a glycolytic enzyme, and the glycolytic enzyme may be a cellulase, as described above.

Oxidative stress can be effectively improved by using a composition containing, as an active ingredient, the enzymatically-treated product of capsular capsular capillaries or the polysaccharide isolated therefrom according to one embodiment.

Figure 1 shows the evaluation results of the ABTS+ radical scavenging activity of an extract of A. celluclast and a polysaccharide derived from the extract.
Figure 2 shows the results of the evaluation of the DPPH radical scavenging activity of the teluclast celluclast extract and the polysaccharide derived from the extract.
Figure 3 shows the results of evaluation of the reducing power of the teluclast extract and the polysaccharide derived from the extract.
Figure 4 shows the results of the evaluation of the cytotoxicity of SSS4 among the polysaccharides derived from only the twisted cap.
Figure 5 shows the results of evaluation of cell viability according to the treatment of each concentration of SSS4 among the polysaccharides derived from the capsular capella extract under UVB irradiation.
6 shows evaluation results of ROS production according to treatment by concentration of SSS4 among polysaccharides derived from capsular capillaries extract during UVB irradiation.

Hereinafter, one or more specific examples will be described in more detail through examples. However, these examples are intended to illustrate one or more specific examples, and the scope of the present invention is not limited to these examples.

Example 1: Material preparation

1-1. Manufacture of pretzel cap enzymatic treatment

The pretzel cap (Sargassum siliquastrum) used in this study was domestically produced, washed thoroughly with water and dried. Phenol solution used in the experiment, H2SO4, HCl, BaCl2, folin-ciocalteu, Na2CO3, AAPH (2,2-azobis (2-amidino-propane) dihydrochloride), DPPH (2, 2-Diphenyl-1-picryl-hydrazyl), potassium ferricyanide, TCA, and FeCl were purchased from Sigma-Aldrich (St. Louis, Missouri, USA), and Dulbeco's modified Eagle's media (DMEM) used for cell experiments. ) is Hyclone Co. (Logan, UT, USA), FBS (Fetal bovine serum) and penicillin, NE-PER Nuclear and Cytoplasmic Extraction Kit (NE-PER Nuclear and Cytoplasmic Extraction Kit) and ECL (Enhanced chemiluminescent) were purchased from Thermo Fisher Scientific Co. (Waltham, MA, USA) was purchased and used. All other analytical reagents used were special grade or HPLC grade reagents.

First, pretzel caps were extracted using celluclast enzyme under optimal enzyme conditions (50 °C, pH 4.5, 24 hours). The extract obtained after the reaction was centrifuged (4000 rpm, 20 minutes), and a final celluclast extract was prepared through filtering and inactivation (95 to 100 ° C, 5 to 10 minutes).

1-2. Production of polysaccharides isolated from the pretzel cap enzymatic treatment

The prepared extract was treated with 100% ethanol and reacted at 4 °C for 12 hours. After the reaction was completed, the primary polysaccharide (SSS1) was obtained by centrifugation (4000 rpm, 20 minutes), and immediately the supernatant was treated with 100% ethanol once more to carry out secondary (SSS2) and tertiary (SSS3) operations in the same manner. , to obtain a quaternary (SSS4) polysaccharide.

1-3. Preparation of dermal keratinocytes (HaCaT)

HaCaT cells, human-derived dermal keratinocytes, were cultured in a DMEM medium containing penicillin/streptomycin (streptomycin 100 unit/mL) and 10% FBS in a 37°C, 5% CO2 incubator and subcultured at 3-day intervals. implemented.

Example 2: Antioxidant effect evaluation method of polysaccharide isolated from the pretzel cap enzyme-treated material

2-1. Method for evaluating free radical scavenging activity and reducing power

(One) Evaluation of ABTS+ radical scavenging activity

ABTS+ 7 mM and potassium persulfate 2.4 mM were mixed and allowed to stand at room temperature for 12 hours to prepare an ABTS+ radical solution. Immediately before use, the molar absorption coefficient (ε = 3.6) was obtained so that the absorbance value at 414 nm was 1.5 Х104 M-1cm-1) was used after diluting with distilled water. A diluted ABTS+ radical solution was added to the positive control (positive control) in which 50 μl and After mixing well, reacting at room temperature for 10 minutes, the absorbance was measured at 414 nm using an ELISA reader.

(2) DPPH radical scavenging activity evaluation

After preparing a DPPH radical solution by mixing DPPH 0.15 mM with methanol, 100 μl of the positive control group treated with the pretzel celluclast extract and polysaccharides derived from the extract (SSS1, SSS2, SSS3, SSS4) and vitamin C (31.3 μg/ml) After mixing well and reacting at room temperature for 30 minutes, the absorbance was measured at 517 nm using an ELISA reader.

(3) reducing power evaluation

Phosphate buffer (0.1 M, pH 6.6/pH 7.0) and 1% potassium ferricyanide, and celuclast extract and polysaccharides derived from the extract (SSS1, SSS2, SSS3, SSS4) and vitamin C ( 31.3 μg/ml) was mixed well with 200 μl of the treated positive control, reacted at 50° C. for 20 minutes, and then 10% TCA was added. The supernatant was separated after centrifugation (3000 rpm, 10 minutes), mixed well with distilled water and 0.1% FeCl3, and absorbance was measured at 700 nm using an ELISA reader.

2-2. Cytotoxicity assessment

To evaluate the cytotoxicity of SSS4, which has the highest free radical scavenging and reducing power on HaCaT cells, MTT assay was performed. HaCaT cells were treated with SSS4 at different concentrations (0, 7.8, 15.6, 31.3, and 62.5 μg/ml) and incubated at 37° C. for 48 hours. The cultured cells were treated with MTT solution (5 mg/ml), incubated at 37 °C for 4 hours, and the color was dissolved from the cells using DMSO. Then, the absorbance was measured at 570 nm using an ELISA reader, and the cell viability was calculated by the following formula.

Survival rate (%) = absorbance of UVB treatment group or UVB and sample treatment group / absorbance of control group * 100

2-3. Evaluation of cytoprotective efficacy of SSS4 in UVB-irradiated HaCaT cells

MTT assay was performed to measure the cytoprotective effect of polysaccharides (SSS1, SSS2, SSS3, and SSS4) isolated from the capsular capsularis on HaCaT cells treated with UVB (Ultraviolet B). HaCaT cells were divided into 1 x 105 cells in a 24 well plate and incubated in each well with 500 μl of DMEM containing 10% FBS, 1% streptomycin and penicillin at 37°C and 5% CO2 for 24 hours. After that, the samples were treated by concentration (25-200 μg/ml), and after 2 hours, stimulated with UVB 40 mJ/cm2 and replaced with FBS-free medium. After the incubation was completed, the supernatant was removed, and 500 μL of DMSO was added to dissolve the formazan precipitate formed by reduction of MTT, and the absorbance was measured at 570 nm using a microplate reader.

2-4. Evaluation of SSS4's intracellular reactive oxygen species (ROS) generation inhibitory activity in UVB-irradiated HaCaT cells

DCFH-DA (2,7'-Dichlorofluorescin diacetate) assay was performed to evaluate intracellular ROS production by polysaccharide treatment isolated from HaCaT cells activated by UVB irradiation. HaCaT cells were divided into 1 x 105 cells in a 24-well plate and incubated in each well with 500 μL of DMEM containing 10% FBS, 1% streptomycin and penicillin at 37°C and 5% CO2 for 24 hours. Thereafter, the separated polysaccharide was treated with different concentrations (25 to 200 μg/ml) and cultured for 2 hours, then stimulated with UVB of 40 mJ/cm2, replaced with FBS-free medium, and further cultured for 1 hour. Then, after treatment with 0.5 mg/ml DCFH-DA solution and incubation for 30 minutes, the amount of intracellular ROS production was measured using a microplate reader at an excitation wavelength of 485 nm/ an emission wavelength of 528 nm. was calculated.

ROS production amount (%) = absorbance of control group or sample treatment group / absorbance of UVB only treatment group * 100

2-5. statistical processing

The experimental results were evaluated for statistical significance using PASW Statistics 19.0 software (SPSS, Chicago, IL, USA), and the significance of the average value between each experimental group was compared using Duncan's test at P < 0.05 level.

Example 3: Antioxidant effect evaluation result of polysaccharide isolated from the pretzel cap enzymatic treatment

3-1. Results of evaluation of free radical scavenging activity and reducing power

As a result of evaluating the free radical scavenging activity and reducing power of the pretzel capella celluclast extract and polysaccharides derived from the extract, as shown in Figs. It was confirmed that In particular, the SSS4 treatment group showed the best activity.

3-2. Cytotoxicity evaluation result

As a result of evaluation of free radical scavenging activity and reducing power, SSS4, which showed the best activity, was selected and further cell experiments were conducted. As a result of evaluating the cytotoxicity of SSS4, it was confirmed that there was no cytotoxicity as SSS4 did not show a difference in survival rate compared to cells that were not treated with anything. Therefore, the following experiment was performed using a concentration that did not show cytotoxicity (FIG. 4).

3-3. Results of evaluation of cytoprotective efficacy of SSS4 in UVB-irradiated HaCaT cells

As a result of evaluating the cytoprotective efficacy of SSS4, in the case of cells irradiated with UVB, the survival rate was significantly decreased compared to cells that were not treated with anything, but it was confirmed that the cell viability increased depending on the treatment concentration of SSS4 (FIG. 5 ).

3-4. Evaluation of the intracellular ROS generation inhibitory activity of SSS4 in UVB-irradiated HaCaT cells

DCF-DA assay was performed to confirm the effect of SSS4 on intracellular ROS production induced by UVB irradiation in HaCaT cells. As a result, in the case of cells irradiated with UVB, it was confirmed that ROS generation was significantly increased compared to cells not treated with anything. However, when SSS4 was treated at different concentrations, intracellular ROS production was significantly reduced (FIG. 6). Therefore, these results indicate that SSS4 increased cell viability by protecting cells by suppressing intracellular ROS production.

So far, the present invention has been looked at with respect to its preferred embodiments. Those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from an illustrative rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent scope will be construed as being included in the present invention.

Claims (11)

A composition for antioxidation comprising, as an active ingredient, enzymatically-treated product of a pretzel cap or a polysaccharide isolated therefrom.
The method of claim 1,
The enzyme is a glycolytic enzyme,
Antioxidant composition.
The method of claim 1,
The enzyme is cellulase,
Antioxidant composition.
A pharmaceutical composition for preventing or treating peripheral vascular disease, allergic dermatitis or skin cancer comprising the antioxidant composition of claim 1 as an active ingredient.
The method of claim 4,
The pharmaceutical composition is an external agent for skin,
pharmaceutical composition
A cosmetic composition comprising an enzymatically treated product of a pretzel cap or a polysaccharide isolated therefrom.
A food composition comprising an enzymatically treated product of a pretzel cap or a polysaccharide isolated therefrom.
A method for producing a polysaccharide derived from capsular caps by treating capsular caps with an enzyme to obtain an extract from caps caps of pretzels.
The method according to claim 8, wherein the step of obtaining the extract is to treat the enzyme for 22 to 26 hours at 40 to 60 ° C., pH is 4 to 5,
Manufacturing method of polysaccharides derived from capsular caps
According to claim 8 or 9,
The enzyme is a glycolytic enzyme,
A method for producing polysaccharides derived from pretzel caps.
According to claim 8 or 9,
The enzyme is cellulase,
A method for producing polysaccharides derived from pretzel caps.

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