KR20220043008A - Antioxidant or Anti-bacterial Composition Using an Extract of Ludwigia epilobioides - Google Patents

Abstract

Disclosed in the present invention is an antioxidant or anti-bacterial composition using an extract of Ludwigia prostrata which has antioxidant activity according to a DPPH method, and has antibacterial activity against Staphylococcus aureus, methicillin-resistant Staphylococcus aureus (MRSA), and Escherichia coli.

Description

Antioxidant or Anti-bacterial Composition Using an Extract of Ludwigia epilobioides

The present invention relates to an antioxidant or antibacterial composition using an extract of Ludwigia epilobioides .

Superoxide radical (O ₂ ㆍ), hydroxyl radical (OH ㆍ), hydrogen peroxide (H ₂ O ₂ ), single oxygen ( ¹ O ₂ ), and the like are collectively called reactive oxygen species (ROS).

These reactive oxygen species are very reactive and react rapidly with biomolecules, causing lipid oxidation, protein denaturation, DNA damage, etc. of biological membranes. It is known to increase the risk of developing disease ( Chem. Res. 14: 393 (1981); Arch. Biophys. 247: 1-11 (1986); Science 221: 1256-1264 (1983)).

Organisms have an antioxidant system to respond to this continuously occurring oxidative stress, and these antioxidant systems are largely enzymatic by superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase, etc. It is divided into a non-enzymatic system by antioxidants such as vitamin A, vitamin C, vitamin E, and antioxidant minerals ingested through the system and diet ( Annu. Rev. Nutr. , 16, pp33-50 (1996)). The enzymatic system and the non-enzymatic system work complementary to each other to detoxify oxidative substances produced in the body.

Antioxidants inhibit or minimize damage to the living body by reactive oxygen species by reacting with active oxygen. Synthetic antioxidants such as PG, Propyl galate) and tertiarybutyl hydroquinone (TBHQ, Teritiarybutyl hydroquinone) and It can be classified as a natural antioxidant, and as synthetic antioxidants are known to cause side effects on carcinogenesis and cardiovascular systems, research on plant-derived antioxidants that are safer and have excellent effects are being actively conducted ( Korean J. Food ). Sci. Technol. 34:889-892 (2002)).

Plants contain antioxidant substances such as polyphenols and flavonoids, so most plant extracts show antioxidant activity. In order to show the , it must be used in large quantities, and consequently, there is a problem in that economical efficiency and practicality are deteriorated due to an increase in the price of raw materials. For this reason, the excavation of plant-derived antioxidants is focused on finding substances with activity comparable to synthetic antioxidants.

TRAP method ( Anal Biochem. , 28, pp300-306. 1989), DPPH method ( Nature 181:1198-1200 (1958)), ABTS method ( Methods Enzymol ) 234 : 279-293 (1994); Clin. Sci. 84: 407-412 (1993); Free Rad. Biol. Med. 20: 933-956 (1996)), ORAC method ( Free Rad. Biol. Med. 18:29-36 (1995)), TOSC method ( Free Rad. Biol. Med. 24(3):480-493 (1998); Toxicol. Appl. Pharmacol. 156(2):96-105 (1999)) , Potassium ferricyanide method (Jpn, J. of Nutr. 44:307-315, 1986), SOD-like activity measurement method ( Eur J Biochem 47:469-474, 1974), such as various methods have been developed.

Meanwhile, bacterial infection is one of the most common and fatal causes of human diseases. In particular, direct or indirect damage caused by pathogenic microorganisms causes many economic, environmental and medical problems. Pathogenic microorganisms are harmful to the human body and environmental pollution due to the use of excessive chemical pesticides on crops in the agricultural industry, the loss due to spoilage during the food distribution process in the food industry, and the emergence of antibiotic-resistant strains due to the misuse of antibiotics. is causing many problems in

In particular, Staphylococcus aureus is a facultatively anaerobic Gram-positive cocci and is the most common causative agent of staphylococcal infections. Staphylococcus aureus forms colonies on the skin and nasal mucosa of healthy people and causes very common infections in the human body due to its high carrier rate. Staphylococcus aureus infection can cause a variety of diseases, ranging from skin infections such as acne, impetigo, and cuts to serious, life-threatening diseases such as pneumonia, sepsis, and endocarditis (J Heart Lung Transplant 2012, 31(11):1199-1206; Microb Pathog 2013;54:26-33; Eur J Clin Microbiol Infect Dis 1994;13(1):50-55).

Among these Staphylococcus aureus, bacteria that are resistant to methicillin, a penicillin-based antibiotic, are called methicillin-resistant Staphylococcus aureus ( MRSA). Recently, it is known that MRSA accounts for more than 70% of Staphylococcus aureus strains in general hospitals or university hospitals in Korea. It is resistant to all β -lactam antibiotics and also has strong resistance to most antibiotics including quinolone antibiotics. N Engl J Med 2005, 352:1436-44; Clin Infect Dis 1994, 19(6):1123-8).

The present invention discloses antioxidant and antibacterial activity of extract

It is an object of the present invention to provide an antioxidant or antibacterial composition using the extract

Other objects or specific objects of the present invention will be set forth below.

As confirmed in the Examples and Experimental Examples below, the present invention has antioxidant activity according to the DPPH method and reducing power, Staphylococcus aureus , methicillin-resistant Staphylococcus aureus. (Methicillin-resistant Staphylococcus aureus , MRSA), and was completed by confirming that it has antibacterial activity against E. coli.

Considering the above, the present invention can be identified as an anti-oxidation composition or antibacterial composition comprising a genus oleifera extract as an active ingredient.

In the present specification, the term "extract of chrysanthemum extract" refers to extracting the stem, leaf, fruit, flower, root, underground part, above-ground part, or mixtures thereof with water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, butanol, etc.) , methylene chloride, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or these It means an extract obtained by leaching using a mixed solvent of In consideration of this, any method such as chilling, reflux, warming, ultrasonic radiation, or supercritical extraction may be applied. In the case of the fractionated extract, a fraction obtained by suspending the extract in a specific solvent and mixing and standing still with a solvent having a different polarity, and adsorbing the crude extract to a column filled with silica gel, etc. It is meant to include the fraction obtained as a mobile phase. In addition, the meaning of the extract includes a concentrated liquid extract or solid extract from which the extraction solvent is removed by methods such as freeze drying, vacuum drying, hot air drying, spray drying, and the like. Preferably, it refers to an extract obtained by using water, ethanol, or a mixed solvent thereof as an extraction solvent, and more preferably an extract obtained by using a mixed solvent of water and ethanol as an extraction solvent.

In addition, as used herein, the term "active ingredient" refers to a component that alone exhibits the desired activity or can exhibit activity together with a carrier that has no activity by itself.

Antioxidant or antibacterial composition of the present invention (hereinafter "composition of the present invention") can be used in any amount (effective amount) ), and a typical effective amount will be determined within the range of 0.001 wt % to 15 wt % based on the total weight of the composition. Herein, the term "effective amount" refers to the intended medical and pharmacological effects, such as antioxidant and antibacterial effects, when the composition of the present invention is administered to a mammal, preferably a human, to which it is applied during the administration period as suggested by a medical professional. It refers to the amount of the active ingredient included in the composition of the present invention. Such an effective amount can be determined empirically within the ordinary ability of one of ordinary skill in the art.

In a specific aspect, the composition of this invention can be grasped|ascertained as a food composition. When the composition of the present invention is identified as a food composition, its use may be understood to inhibit allergic skin hypersensitivity reaction.

The food composition of the present invention can be prepared in any form, for example, beverages such as tea, juice, carbonated drinks, and ion drinks, processed oils such as milk and yogurt, gums, rice cakes, Korean sweets, bread, confectionery, noodles, etc. Foods, tablets, capsules, pills, granules, liquids, powders, flakes, pastes, syrups, gels, jellies, and health functional food preparations such as bars can be manufactured.

In addition, the food composition of the present invention may have any product classification in terms of legal and functional classification as long as it conforms to the enforcement laws at the time of manufacture and distribution. For example, it is a health functional food according to Korea's "Health Functional Food Act", or confectionery, beans, tea, and beverages according to each food type in the Food Ordinance of Korea "Food Sanitation Act" (Ministry of Food and Drug Safety Notification "Food Standards and Specifications") , special purpose food, and the like.

The food composition of the present invention may contain food additives in addition to the active ingredients thereof. Food additives can be generally understood as substances that are added and mixed or infiltrated into food in manufacturing, processing, or preserving food. Food additives with guaranteed safety are limited in terms of ingredients or functions in the Food Additives Ordinance in accordance with the laws of each country that regulates the manufacture and distribution of food (“Food Sanitation Act” in Korea). In the Korean Food Additives Code (“Food Additive Standards and Specifications” announced by the Ministry of Food and Drug Safety), food additives are classified into chemically synthetic products, natural additives, and mixed preparations in terms of ingredients. It is divided into agents, preservatives, emulsifiers, acidulants, thickeners, etc.

The sweetener is used to impart appropriate sweetness to food, and both natural and synthetic ones may be used in the composition of the present invention. Preferably, a natural sweetener is used. Examples of the natural sweetener include sugar sweeteners such as corn syrup solids, honey, sucrose, fructose, lactose, and maltose.

Flavoring agents may be used to improve taste or aroma, and both natural and synthetic ones may be used. Preferably, it is a case where a natural thing is used. In the case of using a natural product, the purpose of nutritional enhancement in addition to flavor may be concurrently used. The natural flavoring agent may be obtained from apples, lemons, tangerines, grapes, strawberries, peaches, or the like, or obtained from green tea leaves, honeysuckle, bamboo leaves, cinnamon, chrysanthemum leaves, jasmine, and the like. In addition, those obtained from ginseng (red ginseng), bamboo shoots, aloe vera, and ginkgo can be used. The natural flavoring agent may be a liquid concentrate or a solid extract. Optionally, a synthetic flavoring agent may be used, and the synthetic flavoring agent may be an ester, an alcohol, an aldehyde, a terpene, or the like.

As a preservative, sodium calcium sorbate, sodium sorbate, potassium sorbate, calcium benzoate, sodium benzoate, potassium benzoate, EDTA (ethylenediaminetetraacetic acid), etc. can be used, and as an emulsifier, acacia gum, carboxymethylcellulose, xanthan gum, Pectin, etc. are mentioned, and acidulant, malic acid, fumaric acid, adipic acid, phosphoric acid, gluconic acid, tartaric acid, ascorbic acid, acetic acid, phosphoric acid, etc. can be used as an acidulant. The acidulant may be added so that the food composition has an appropriate acidity for the purpose of inhibiting the growth of microorganisms in addition to the purpose of enhancing the taste.

As a thickening agent, a suspending agent, a settling agent, a gel former, a bulking agent, etc. can be used.

The food composition of the present invention may contain, in addition to the food additives described above, physiologically active substances or minerals known in the art for the purpose of supplementing and reinforcing functionality and nutrition and guaranteed stability as food additives.

Examples of such physiologically active substances include catechins contained in green tea and the like, vitamins such as vitamin B1, vitamin C, vitamin E, and vitamin B12, tocopherol, dibenzoylthiamine, and the like. Minerals include calcium preparations such as calcium citrate, magnesium stearate Magnesium preparations, such as iron preparations, such as iron citrate, chromium chloride, potassium iodide, selenium, germanium, vanadium, zinc, etc. are mentioned.

In the food composition of the present invention, the food additives as described above may be included in an appropriate amount to achieve the purpose of the addition according to the product type.

In relation to other food additives that may be included in the food composition of the present invention, reference may be made to the Food Ordinance of each country or the Food Additives Code.

The composition of the present invention may be regarded as a pharmaceutical composition in another specific embodiment.

The pharmaceutical composition of the present invention may be prepared as an oral dosage form or a parenteral dosage form according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. Here, the route of administration may be any suitable route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and two or more routes may be used in combination. An example of the combination of two or more routes is a case in which two or more formulations of drugs according to the route of administration are combined. For example, one drug is first administered by an intravenous route and the other drug is secondarily administered by a local route.

Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specifically, reference may be made to the pharmacopoeia of each country including the "Korea Pharmacopoeia".

When the pharmaceutical composition of the present invention is prepared as an oral dosage form, powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers according to methods known in the art together with a suitable carrier It can be prepared in a formulation such as Examples of suitable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, and xylitol, starches such as corn starch, potato starch, wheat starch, cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, Cellulose such as hydroxypropylmethylcellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyol, vegetable oil, ethanol, glycerol and the like. In the case of formulation, an appropriate binder, lubricant, disintegrant, colorant, diluent, etc. may be included as needed. Suitable binders include starch, magnesium aluminum silicate, starch paste, gelatin, methylcellulose, sodium carboxymethylcellulose, polyvinylpyrrolidone, glucose, corn sweetener, sodium alginate, polyethylene glycol, wax, and the like, and lubricants include sodium oleate. , sodium stearate, magnesium stearate, sodium benzoate, sodium acetate, sodium chloride, silica, talcum, stearic acid, its magnesium salt and calcium salt, polyethylene glycol, etc., and the disintegrant includes starch, methyl cellulose, agar, bentonite, xanthan gum, starch, alginic acid or its sodium salt and the like. Examples of the diluent include lactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine, and the like.

When the pharmaceutical composition of the present invention is prepared for parenteral use, it may be formulated in the form of injections, transdermal administrations, nasal inhalants and suppositories together with suitable carriers according to methods known in the art. When formulated as an injection, an aqueous isotonic solution or suspension may be used as a suitable carrier, and specifically, PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or isotonic solution such as 5% dextrose may be used. . When formulated for transdermal administration, it can be formulated in the form of an ointment, a cream, a lotion, a gel, an external solution, a pasta agent, a liniment agent, an air roll, and the like. In the case of nasal inhalants, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide, and the like. witepsol), tween 61, polyethylene glycols, cacao fat, laurin fat, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene stearate, sorbitan fatty acid esters, and the like can be used.

Specific formulations of pharmaceutical compositions are known in the art, and reference may be made to, for example, Remington's Pharmaceutical Sciences (19th ed., 1995). This document is considered a part of this specification.

A preferred dosage of the pharmaceutical composition of the present invention is in the range of 0.001 mg/kg to 10 g/kg per day, preferably 0.001 mg/kg to 1 g, depending on the patient's condition, weight, sex, age, severity of the patient, and the route of administration. It can be in the range /kg. Administration may be performed once or divided into several times a day. These dosages should not be construed as limiting the scope of the invention in any respect.

In another specific embodiment, the composition of the present invention can be identified as a cosmetic composition.

Even when the composition of the present invention is identified as a cosmetic composition, the cosmetic composition can be classified into any product in terms of its use or by law, and specifically functional cosmetics, non-functional products having uses such as improvement of skin troubles and improvement of atopic dermatitis It may be general cosmetics and the like. In terms of product form, it may take any product form, and specifically, solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax It can take the form of products such as foundation and spray. In the specific product form, it may be in the form of a flexible lotion, a nourishing lotion, a nourishing cream, a massage cream, an essence, an eye cream, a cleansing cream, a cleansing foam, a cleansing water, a pack, a spray, or a powder.

The cosmetic composition of the present invention may include components commonly used in cosmetic compositions in addition to the active ingredient, for example, conventional adjuvants such as stabilizers, solubilizers, surfactants, vitamins, pigments and fragrances, and carriers.

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 may be used as a carrier component. can

When 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. In particular, in the case of a spray, additional chlorofluorohydrocarbon, propane /may contain propellants such as butane or dimethyl ether.

When the formulation of the present invention is a solution or emulsion, a solvent, solubilizer or emulsifier is used as a carrier component, specifically water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylglycol oil, glycerol aliphatic ester, polyethylene glycol, fatty acid ester of sorbitan, etc. 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, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester, polyoxyethylene sorbitan ester, microcrystals Adult cellulose, aluminum metahydroxide, bentonite, agar, etc. can be used.

When the formulation of the present invention is a surfactant-containing cleansing agent, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyltaurate, sarcosinate, fatty acid amide as carrier components Ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative or ethoxylated glycerol fatty acid ester and the like may be used.

The cosmetic composition of the present invention can be prepared according to the method for preparing a cosmetic composition conventionally performed in the art, except that it contains the active ingredient exhibiting antioxidant and antibacterial activity.

As described above, according to the present invention, it is possible to provide a composition for anti-oxidation or antibacterial using the extract of chrysanthemum.

The composition of the present invention may be commercialized as food, drug or cosmetic.

1 and 2 are experimental results for the antioxidant activity of the extract of yew needles.
3 to 4 are experimental results for the antibacterial activity of the extract of yew needles.

Hereinafter, the present invention will be described with reference to Examples and Experimental Examples. However, the scope of the present invention is not limited to these Examples and Experimental Examples.

<Example> Preparation of leaf extract and stem extract

After adding 30 times the weight of distilled water (at 120° C. for 2 hours) to the dry powder of dandelion leaves, hot water extraction was performed, followed by filtration, removal of the extraction solvent, and freeze-drying to prepare a quince needle leaf extract.

The extract of the genus needles stems was prepared by hot water extraction in the same manner as above.

<Experimental Example> Antioxidant activity and antibacterial activity

<Experimental Example 1> Antioxidant activity experiment

<Experimental Example 1-1> Antioxidant activity experiment by DPPH assay

The antioxidative activity of the extracts of chrysanthemums according to the DPPH (1,1-diphenyl-2-picryl hydrazyl) method was confirmed as follows.

DPPH radical scavenging activity is a method of measuring the scavenging ability of a sample for free radicals generated by DPPH in ethanol. 0.1 ml of each extract diluted to a certain concentration in 0.1 ml of a 0.2 mM DPPH solution dissolved in methanol was added and mixed for 10 seconds. Then, after blocking the light and reacting at room temperature for 30 minutes, the absorbance was measured at a wavelength of 517 nm using a spectrophotometer. Referring to FIG. 1 , it can be seen that both the leaf extract and the stem extract show DPPH scavenging activity in a concentration-dependent manner.

<Experimental Example 1-2> reducing power measurement experiment

The reducing power was measured using the potassium ferricyanide method. To 1.0 ml (1 mg/ml) of each extract, 2.5 ml of 200 mM phosphate buffer (pH 6.6) and 1% potassium ferricyanide solution were sequentially added and stirred, followed by reaction at 50° C. for 20 minutes. To the reaction solution, 2.5 ml of 10% trichloroacetic acid was added, centrifuged at 3,500 rpm for 10 minutes, and 2.5 ml of distilled water and 0.5 ml of ferric chloride solution were added to 2.5 ml of the supernatant and mixed, followed by a multimode microplate reader (Varioskan LUX, ThermoFisher Scientific, Inc.). (Co. MA, USA) was used to measure the absorbance at 700 nm. The analysis result was shown in FIG. 2 as an absorbance of 700 nm. Referring to FIG. 2 , it can be seen that the reducing power is increased in a concentration-dependent manner for both the genus oleifera leaf extract and the stem extract, as in the result of FIG. 1 .

<Experimental Example 2> Antibacterial activity test

<Experimental Example 2-1> Antibacterial activity test according to the disc diffusion assay

The strains used to confirm the antibacterial effect are Gram-positive bacteria, Staphylococcus aureus ATCC 29213, as shown in Table 1 below; Two methicillin-resistant Staphylococcus aureus (MRSA) and one gram-negative Escherichia coli were used, and the medium was cultured at 37°C using LB broth and LB agar. Escherichia coli, a gram-negative bacterium, was purchased from Gachon University, Staphylococcus aureus, a gram-positive bacterium, a strain acquired from the Korea Microbial Conservation Center was used, and MRSA was purchased and used from Gachon University Gil Hospital.

The antibacterial activity of the extracts of chrysanthemum needles was confirmed by a disc diffusion assay using a paper disc. For strain inoculation, the culture medium was transplanted into 3 ml of sterilized culture medium through a high-temperature high-pressure sterilizer, and cultured with shaking at 37° C. for 24 hours. The cultured strain was diluted to 1 x 10 ⁵ cfu/ml and spread evenly on an agar plate. After inoculating the extracts by concentration on sterilized paper discs, lightly placing them on the surface of the medium, and culturing at 37°C for 24 hours, it was checked whether or not a clear zone was generated.

The results are shown in Figures 3 (leaf) and 4 (stem) of the growth inhibition ring (clear zone) of each of the leaf extract and stem extract, while the growth inhibition is shown in Table 2 (leaf) and Table 3 (stem) below. The size of the ring was measured and shown.

Both the leaf extract and the stem extract showed antibacterial activity against three microorganisms, and the leaf extract was particularly excellent in antibacterial activity. 1,2-Hexanediol was used as a positive control.

<Experimental Example 2-2> Minimum inhibitory concentration (MIC) measurement experiment

To measure the minimum growth inhibitory concentration (MIC) of the extract for the test strain, dilute the extract by 1/2 with sterile LB broth until the minimum concentration is 0.004%, and then put 20 μl into a 96 well plate, and then each strain 1 x 10 ⁵ cfu/ml 180 μl was added. After culturing for 24 hours in a 37 °C incubator, absorbance was measured at a wavelength of 595 nm through an absorbance meter, and the concentration of the extract in which the inoculated strain was not grown was used as the MIC.

The results are shown in Table 4 below. The minimum inhibitory concentration of the genus leaf extract was 1 mg/mL, and the minimum inhibitory concentration of the genus oleifera stem extract was 2 mg/mL.

Claims (7)

A composition for antioxidants comprising the extract of chrysanthemum needles as an active ingredient.
The method of claim 1,
The extract is a composition characterized in that the extract of water, ethanol, or a mixed solvent thereof of the leaves or stems of the cypress needles.
An antibacterial composition comprising an extract of chrysanthemum needles as an active ingredient.
4. The method of claim 3,
The antibacterial is Staphylococcus aureus ( Staphylococcus aureus ), methicillin-resistant Staphylococcus aureus (Methicillin-resistant Staphylococcus aureus , MRSA), and the composition characterized in that the antibacterial activity against E. coli.
4. The method of claim 3,
The extract is a composition characterized in that the extract of water, ethanol, or a mixed solvent thereof of the leaves or stems of the cypress needles.
4. The method of claim 3,
The composition is a food composition, characterized in that the composition.
4. The method of claim 3,
The composition is a cosmetic composition, characterized in that the composition.

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