KR20240020237A - Composition for antibacterial and anti-inflammatory comprising extract of coastal hogfennel, and composition for preventing or improving asthma, eczema and rhinitis comprising the same - Google Patents

Abstract

The present invention relates to an antibacterial and anti-inflammatory composition containing an extract of Prickly pear perilla and a composition containing the same for preventing or improving asthma, eczema and rhinitis.
The composition according to one embodiment of the present invention exhibits excellent antibacterial and anti-inflammatory effects and has no side effects. Additionally, the present invention provides various formulations of food compositions and other external agents that prevent or improve allergic and seasonal diseases including asthma, eczema, and rhinitis.

Description

An antibacterial and anti-inflammatory composition containing an extract of P. persimmon sinensis and a composition containing the same for preventing or improving asthma, eczema and rhinitis AND RHINITIS COMPRISING THE SAME}

The present invention relates to an antibacterial and anti-inflammatory composition containing an extract of Perilla sinensis and a composition containing the same for preventing or improving asthma, eczema and rhinitis. More specifically, the present invention can provide various formulations of food compositions and other external agents that exhibit excellent antibacterial and anti-inflammatory effects and prevent or improve allergic and seasonal diseases including asthma, eczema, and rhinitis.

Allergy generally refers to a phenomenon that causes harmful damage to the body by a reaction caused by an immunological mechanism due to an allergen, a foreign substance.

These allergic reactions occur through mast cells. Mast cells are widely distributed in organs throughout the body, such as the skin, respiratory tract, mucosa of the gastrointestinal tract, lymphatic vessels, blood vessels, and brain, and are known as the causative cells of allergic reactions.

Allergic reactions are caused by IgE produced to combat allergens such as house dust mites, pollen, animal dander, mold, ovalbumin, milk protein, and peanuts, acting on mast cells (Wills-Karp M., et al., Science, 282, pp 2258-61, 1998; Grunig G., et al., Science, 282, pp 2261-2263, 1998).

Looking at this specifically, when an allergen invades the living body, the allergen is first recognized by antigen-presenting cells, and these antigen-presenting cells differentiate Th0 into Th2 cells through allergen presentation in the presence of IL-4. These differentiated Th2 cells secrete cytokines such as IL-4, IL-5, and IL-13, which act on B cells to induce the production of IgE.

The IgE produced in this way binds to FcεRⅠ, a high-affinity IgE receptor on mast cells. When exposed to the same allergen again, the allergen binds to IgE bound to FcεRⅠ on mast cells, causing cross-linking of these receptors, which causes signaling. As a result, a series of signaling reactions proceed.

Through this signaling reaction, degranulation of mast cells is induced and histamine, heparin, bradykinin, triptase, and chymase are released, while cell membrane phospholipids are released. In the body, arachidonic acid metabolites such as leukotrien and prostaglandin are newly synthesized and released. These substances cause dilation of blood vessels, increased permeability, stimulation of nerve endings, and secretion of mucus. It causes a series of allergic reactions, including hyperactivity and contraction of smooth muscles.

Since allergic reactions are caused by mast cell degranulation, substances that inhibit mast cell degranulation have potential as a treatment for allergic diseases (Choi Seon-pil et al., J. Korean Soc. Appl. Biol. Chem. 48(4), 315-321 (2005)).

Meanwhile, Staphylococcus aureus and Staphylococcus epidermidis are common skin bacteria, and along with Streptococcus pyogenes, they cause purulent infections, cellulitis, or general inflammation on the skin. This is a representative strain that can cause.

Antibacterial agents such as tetracycline, erythromycin, and azelaic acid are used for these skin infections, but side effects such as emergence of resistant strains, human toxicity such as skin hypersensitivity reaction, and photosensitivity are reported. Recently, research on antibacterial agents using natural products has been actively conducted. Therefore, a composition is needed as a natural product-based antibacterial agent that can be used for a long period of time without side effects.

Meanwhile, some of the substances that play an important role in the process of maintaining homeostasis of living organisms are physiologically active substances derived from various living organisms. To date, much research has been conducted on numerous biologically active substances, and among them, research on antibacterial substances isolated from microorganisms, plants, or animals can be said to be a very important area in the fields of life science and medicine.

All living things are known to produce antibacterial and antifungal substances for survival, and much research is being conducted on microorganisms that produce antibacterial and antifungal substances. Research to develop antibacterial or antifungal agents has been attempted for a long time.

Antibacterial substances are substances that kill microorganisms, have low toxicity to humans or animals, and have selective toxicity that is not inactivated by enzymes in the body. They mainly involve the replication of DNA, the transcription and decoding of genetic information, and the use of electronic energy. It is effective through a mechanism that inhibits the growth of microorganisms by inhibiting transport and cell wall biosynthesis.

The main antibacterial substances developed to date can be divided into those of natural origin, such as microorganisms and plants, and those that are chemically synthesized. The first antibacterial substance isolated from nature was penicillin, discovered by Alexander Fleming in 1928, and has since been classified as either naturally derived or chemically synthesized. Many chemically synthesized antibacterial substances are used to treat diseases caused by harmful bacteria, but recently, due to increasing resistance to chemically synthesized antibacterial substances, there is an increasing interest in developing antibacterial drugs using naturally derived antibacterial substances. am.

Technology related to naturally derived antibacterial or antifungal agents, including Sophora ginseng extract (KR 10-1994-0023496 A), grapefruit extract (patent application KR 10-1993-0006320 A), mixed extracts of fennel, fennel, sesame seeds, camphor plants, and cloves. Extracts from various plants, such as (patent application KR 10-2003-0035851 A), green tea polyphenol and tea tree oil (KR 10-2003-0090322 A), and Sesbania grandiflora extract (KR 10-2011-0027735 A). It has been reported so far that the fraction has antibacterial activity.

KR 10-1994-0023496 A KR 10-1993-0006320 A KR 10-2003-0035851 A KR 10-2003-0090322 A KR 10-2011-0027735 A

The purpose of the present invention is to provide a composition having antibacterial and anti-inflammatory activities using natural product-based extracts.

The purpose of the present invention is to provide a composition that can exhibit antibacterial and anti-inflammatory activity while being used for a long period of time without the problem of side effects using a composition based on plant extracts.

Another object of the present invention is to enable the preparation of compositions for preventing or improving allergic and seasonal diseases such as asthma, eczema and rhinitis in various formulations using the composition.

In order to achieve the above object, an antibacterial and anti-inflammatory composition according to an embodiment of the present invention contains an extract of Perilla perilla.

The extract may be extracted using an extraction solvent selected from the group consisting of water, alcohols having 1 to 6 carbon atoms, and mixtures thereof.

The composition includes Gujeolcho extract; tangerine peel extract; It may further include any one selected from the group consisting of Dalgae extract and mixtures thereof.

A functional food for antibacterial and anti-inflammatory purposes according to another embodiment of the present invention includes the composition.

An external antibacterial and anti-inflammatory skin preparation according to another embodiment of the present invention includes the composition.

A composition for preventing and improving allergic diseases according to another embodiment of the present invention may include the extract.

A composition for preventing or improving asthma, eczema and rhinitis according to another embodiment of the present invention may include the extract.

Hereinafter, the present invention will be described in more detail.

Inflammation as referred to in the present invention is a type of in vivo response to damage or infection of a specific tissue, and the main mediator is an immune cell. Depending on the damaged or infected area of the tissue, it can be divided into acute inflammation and chronic inflammation. inflammation). When the damaged or infected area of the tissue is small or temporary, it is resolved by acute inflammation, but when the inflamed area is large or chronically infected, it is not resolved by acute inflammation and progresses to chronic inflammation. Chronic inflammation can progress into chronic inflammatory disease.

“Immune disease” as used in the present invention refers to a disease caused by an excessive response of the immune system in the human body, and representative examples include allergic diseases. However, since the inflammatory reaction is ultimately caused by the reaction of the immune system in the body, inflammatory disease and immune disease can be used as terms to refer to similar ranges in a broad sense, and are not used as clearly distinct terms within this specification. , will be used interchangeably.

In addition, the immune disease refers to a disease that causes problems when a specific immune response occurs, and is not limited thereto, but preferably includes autoimmune disease, transplant rejection, and graft-versus-host disease. Autoimmune diseases include Crohn's disease and erythema. Disease, atopy, rheumatoid arthritis, Hashimoto's thyroiditis, pernicious anemia, Addison's disease, type 1 diabetes, lupus, chronic fatigue syndrome, fibromyalgia, hypothyroidism and hyperthyroidism, scleroderma, Behçet's disease, inflammatory bowel disease, multiple sclerosis, severe It may be myasthenia gravis, Meniere's syndrome, Guilian-Barre syndrome, Sjogren's syndrome, vitiligo, endometriosis, psoriasis, vitiligo, systemic scleroderma, asthma or ulcerative colitis.

In addition, asthma, eczema, and rhinitis as referred to in the present invention are representative examples of the above-mentioned immune disease, allergic disease, or seasonal disease, and the immune disease, allergic disease, or seasonal disease as referred to in the present invention is defined to include asthma, eczema, and rhinitis. .

The term “inflammatory disease” used in the present invention refers to a disease caused by an inflammatory reaction derived from external or internal substances, and representative examples include neuroinflammatory disease and arthritis.

An antibacterial and anti-inflammatory composition according to an embodiment of the present invention contains an extract of Perilla perilla.

Peucedanum japoincum Thunberg is mainly distributed in well-developed coastal areas in the southern region, but also grows in crevices in rocks on rocky coasts. According to literature, there is a record of it being distributed in coastal sand dunes on the east and west coasts. The leaves are thick, split into three, irregularly serrated, and the leaves and roots are fragrant. The leaves are small, about 10cm tall, and the flower stems are over 10cm tall. White flowers bloom starting in the fall, and after the flowers fade, the flower stems remain dead.

When the above-mentioned perilla seed extract is used, it can exhibit excellent antibacterial activity and salt-containing effects. Specifically, the extract increases immune activity by increasing cytokine production of TNF-α and IL-12, thereby achieving the effect of preventing and improving allergic diseases.

The composition includes Acinetobacter genus, Alcaligenes genus, Enterobacter genus, Bacillus genus, Escherichia genus, Listeria genus, Proteus genus, Providencia genus, Pseudomonas genus, Salmonella genus, Staphylococcus genus, Candida genus, Saccharomyces genus, Aspergillus genus, for microorganisms selected from the group consisting of Cladosporium, Paecilomyces, Penicillium, Trichothecium, and Klebsiella pneumoniae species. Excellent antibacterial effect.

Cytokines are protein immunomodulators secreted from immune cells and bind to specific receptors during autocrine signaling, paracrine signaling, and endocrine signaling to produce immune response. involved in There are various types of cytokines involved in cell proliferation, differentiation, apoptosis, or wound healing, and many are particularly involved in immunity and inflammation.

Among these, the most important immune functions are performed by a group of cytokines known as interleukin (Interleukin-1, -6, -12), interferon gamma (IFN-γ), and tumor necrosis factor (TNF).

Interleukin (IL) is secreted from white blood cells and is involved in immune system regulation. Currently, more than 30 types are known. Interleukin is a substance that plays an important role in the body's defense function by regulating the immune response by acting on various stages of the immune response. Its types include IL-1, 6, and 12.

Among these, IL-12 is a cytokine composed of p35 and p40 proteins, and has a very important immunological function in linking innate and acquired immunity. It is also involved in the removal of endogenous antigens by various effector cells, such as macrophages. ), induces the activation of NK cells and T-cells. NK cells and T-cells activated by stimulation of IL-12 have the characteristics of being major factors mediating the early innate immune response by stimulating the production of IFN-γ. In addition, it is the most important mediator of cellular immunity because it induces the immune system of T-cells to become Th-1-oriented.

Interferon (IFN) functions to prevent the proliferation of viruses or control cell proliferation and also plays an important function in the immune system. Interferons come in alpha (α), beta (β), gamma (γ), and omega (ω) types. Interferon-α is mainly secreted by macrophages, and interferon-β is mainly secreted by fibroblasts. Interferon-γ is secreted by Th1 cells and NK cells, and interferon-ω is mainly secreted by trophoblasts or trophoblasts. In particular, interferon-γ (IFN-γ) is produced by T lymphocytes and macrogranulocytes and is a substance that induces the killing of phagocytosed microorganisms or antigens. In addition, it has various immunomodulatory effects and activates macrophages.

Tumor necrosis factor (TNF) is a cell signaling protein related to inflammatory response and is a cytokine that induces acute phase response. Tumor necrosis factor is divided into tumor necrosis factor-α (TNF-α) and tumor necrosis factor-β (TNF-β), also called lymphotoxin, but tumor necrosis factor generally refers to tumor necrosis factor-α. .

Therefore, in the case of the immune function improvement effect of the antibacterial, anti-inflammatory, antiviral, and immune function improvement composition of the present invention, natural products without human toxicity and side effects are used, and TNF-α and IL-12, which activate the immune response, are used. It increases the production of cytokines.

Therefore, when using the extract, the effect of preventing and improving allergic diseases or immune diseases including asthma, eczema, and rhinitis can be achieved.

The term 'extract' used in this specification has the meaning commonly used in the art as a crude extract, as described above, but in a broad sense also includes fractions obtained by additional fractionation of the extract. In other words, plant extracts include not only those obtained using the above-described extraction solvent, but also those obtained by additionally applying a purification process. For example, fractions obtained by passing the extract through an ultrafiltration membrane with a certain molecular weight cut-off value, separation by various chromatographs (designed for separation according to size, charge, hydrophobicity, or affinity), etc. Fractions obtained through various purification methods are also included in the plant extract of the present invention.

The extract used in the present invention can be prepared in powder form by additional processes such as reduced pressure distillation and freeze drying or spray drying.

The extract may be extracted using an extraction solvent selected from the group consisting of water, alcohols having 1 to 6 carbon atoms, and mixtures thereof.

The pulverized product of the extract can be eluted with a polar solvent such as an alcohol having 1 to 6 carbon atoms such as water, ethanol, or methanol, or a mixed solvent having a mixing ratio of alcohol and water of 1:0.1 to 1:10. For more details, Alternatively, water can be used as an extraction solvent. At this time, the extraction temperature may be 10°C to 100°C, preferably the extract may be extracted at room temperature.

The extraction solvent can be used in an amount of 2 to 50 times, preferably 2 to 20 times, based on the weight of the sample. For extraction, the sample may be left in the extraction solvent for 1 to 72 hours, specifically 1 to 24 hours.

It may be a result obtained by concentrating the filtered extract under reduced pressure with a vacuum rotary concentrator, but is not limited thereto, and includes all extracts, diluted or concentrated extracts, dried products obtained by drying the extracts, and crude or purified products thereof.

The composition includes Gujeolcho extract; tangerine peel extract; It may further include any one selected from the group consisting of Dalgae extract and mixtures thereof.

Preferably, the composition includes Gujeolcho extract; It may include tangerine peel extract and moongave extract. In the case of the above mixed composition, antibacterial and anti-inflammatory activities can be further improved.

More preferably, the composition includes an extract of Peach oil sprouts, and 1 to 10 parts by weight of Gujeolcho, based on 100 parts by weight of the extract of Peach oil sprouts; It may include 40 to 80 parts by weight of tangerine peel and 1 to 10 parts by weight of tangerine peel.

In the case of the above mixing range, a synergistic effect can be achieved due to the interaction of the active ingredients contained in each extract, and excellent antibacterial and anti-inflammatory activities can be achieved with a smaller amount. Accordingly, the problem of side effects or problems caused by long-term use can be resolved.

More preferably, the composition may further include an extract of the herbaceous plant extract or an extract of the chinensis chinensis extract. When the extract is further included, the effect of additionally increasing overall antibacterial and anti-inflammatory activity can be achieved through different mechanisms of the included active ingredients.

In particular, the composition includes a Peach oil sprout extract, and based on 100 parts by weight of the Peach oil sprout extract, 1 to 10 parts by weight of Gujeolcho; 40 to 80 parts by weight of tangerine peel; It may be included in the amount of 1 to 10 parts by weight of Dalgae, 0.1 to 1 part by weight of Dalgae, and 10 to 20 parts by weight of nasturtium.

In the case of the above mixing range, additional synergistic activity is maximized and excellent antibacterial and anti-inflammatory activities can be achieved. Therefore, excellent effects can be achieved over a long period of time with a small amount without problems such as cytotoxicity.

In addition, within the above range, it can be manufactured into various formulations, easily formulated into products for improving immunity, products for preventing and improving allergic diseases, etc., and excellent effects can be achieved with a small amount.

A functional food for antibacterial and anti-inflammatory purposes according to another embodiment of the present invention includes the composition.

The food composition includes health functional foods or general foods.

The health functional food is not particularly limited as long as it can be consumed to improve antibacterial, anti-inflammatory, antiviral and immune activity. When using the health functional food composition of the present invention as a food additive, the health functional food composition may be added as is or used together with other foods or food ingredients, and may be used appropriately according to conventional methods. The active ingredient can be used appropriately depending on its purpose of use (prevention or improvement). Generally, when manufacturing a food or beverage, the health functional food composition of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on the raw materials. However, in the case of long-term intake for health purposes, the amount may be below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

There are no particular restrictions on the types of health functional foods. The health functional food composition of the present invention can be manufactured into food, especially functional food. The functional food of the present invention includes ingredients commonly added during food production, and includes, for example, proteins, carbohydrates, fats, nutrients and seasonings. For example, when manufacturing a drink, natural carbohydrates or flavoring agents may be included as additional ingredients in addition to the active ingredient. The natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), oligosaccharides, polysaccharides (e.g., dextrins, cyclodextrins, etc.), or sugar alcohols (e.g., , xylitol, sorbitol, erythritol, etc.) are preferred. The flavoring agent may be a natural flavoring agent (e.g., thaumatin, stevia extract, etc.) or a synthetic flavoring agent (e.g., saccharin, aspartame, etc.).

In addition to the above health functional food composition, various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonic acid. It may further contain carbonating agents used in beverages.

Taking the general processed food as an example, it is possible to manufacture processed food that contains the extract and can enhance antibacterial, anti-inflammatory, antiviral, and immune function improvement effects. These processed foods include, for example, snacks, beverages, alcoholic beverages, fermented foods, canned foods, processed milk foods, processed meat foods, noodles, etc. Confectionery includes biscuits, pies, cakes, bread, candy, jellies, gum, cereals, etc. Beverages include drinking water, carbonated beverages, functional ionic beverages, functional ionic beverages, juices (e.g., apple, pear, grape, aloe, tangerine, peach, carrot, tomato juice, etc.), sikhye, etc. Alcoholic beverages include sake, whiskey, soju, beer, liquor, and fruit wine. Fermented foods include soy sauce, soybean paste, and red pepper paste. Canned food includes canned seafood (e.g., canned tuna, mackerel, saury, canned conch, etc.), canned livestock products (canned beef, pork, chicken, turkey, etc.), and canned agricultural products (canned corn, peaches, canned file apple, etc.). Milk processed foods include cheese, butter, yogurt, etc. Processed meat products include pork cutlet, beef cutlet, chicken cutlet, and sausage. Includes sweet and sour pork, nuggets, neobiani, etc. Includes noodles such as raw noodles in sealed packages. In addition, the composition can be used in retort foods, soups, etc.

An external antibacterial and anti-inflammatory skin preparation according to another embodiment of the present invention includes the composition.

Formulations of the skin external preparation include, for example, solutions, gels, solid or pasty anhydrous products, emulsions obtained by dispersing the oil phase in the water phase, suspensions, microemulsions, microcapsules, microgranules, or ionic (liposome) and non-ionic types. It may be provided in the form of a vesicular dispersant, cream, skin, lotion, powder, ointment, spray or conceal stick. Additionally, it can be prepared in the form of foam or in the form of an aerosol composition further containing compressed propellant.

In addition, the skin external preparations include cosmetics, such as astringent lotions, softening lotions, nourishing lotions, various creams, essences, packs, foundations, cleansing products, face washes, soaps, treatments, or beauty essences. Specific formulations include skin lotion, skin softener, skin toner, astringent, lotion, milk lotion, moisture lotion, nutritional lotion, massage cream, nutritional cream, moisture cream, hand cream, essence, nutritional essence, pack, soap, shampoo, and cleansing foam. , can be manufactured into formulations such as cleansing lotion, cleansing cream, body lotion, body cleanser, emulsion, press powder, loose powder, or eye shadow. However, it is not limited to this.

A composition for preventing or improving allergic diseases according to another embodiment of the present invention may include the composition.

The allergic disease may include asthma, eczema, and rhinitis.

The present invention provides a composition having antibacterial and anti-inflammatory activities using natural product-based extracts.

The present invention provides a composition that uses a composition based on plant extracts to exhibit antibacterial and anti-inflammatory activity while being used for a long period of time without the problem of side effects.

Another embodiment of the present invention allows the preparation of compositions for preventing or improving allergic and seasonal diseases such as asthma, eczema and rhinitis in various formulations using the composition.

Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement it. However, the present invention may be implemented in many different forms and is not limited to the embodiments described herein.

[Preparation example: Preparation of extract]

1. Preparation of perilla perilla extract

After washing and drying the perilla seedlings, they were pulverized. The pulverized product was subjected to hot water extraction, filtered, and concentrated to prepare AE perilla extract (AE).

2. Preparation of other extracts

In the same manner as the above-mentioned perilla plant extract, hot water extraction of Gujeolcho, tangerine peel, Dalgae, Gaebululpul, and Nasturtium was performed to produce Gujeolcho extract (BE), Tangerine peel extract (CE), Dalgae extract (DE), Gaebululpul extract (EE), Nasturtium chinensis extract (FE) was prepared.

3. Preparation of mixed formulation

In order to evaluate the synergistic activity through the mixed composition of each extract in addition to the activity of each extract, a mixed formulation was prepared by mixing the AE to FE in the composition shown in [Table 1] below.

M1 M2 M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 A.E. 100 100 100 100 100 100 100 100 100 100 100 100 BE 100 - 0.1 One 5 10 20 5 5 5 5 5 C.E. - 100 20 40 60 80 100 60 60 60 60 60 DE - - 0.1 One 5 10 20 5 5 5 5 5 EE - - - - - - - 0.01 0.1 0.5 One 2 F.E. - - - - - - - One 10 5 20 30

(Unit: parts by weight)

[Experimental example: activity evaluation]

1. Antibacterial activity evaluation

To evaluate the antibacterial activity of each extract and mixed formulation, Staphylococcus aureus KCTC1928 (SA), Escherichia coli (EC), Pseudomonas aeruginosa (PA), and methicillin-resistant Staphylococcus aureus (MRSA) were tested. aureus) was used as a test strain.

The test strain was cultured using a medium (Muller hinton broth) at a temperature of 37°C until the McFarland turbidity reached 0.5, and then used in an antibacterial test.

Evaluation of antibacterial activity was performed using the paper disk diffusion method (Toama et al., 1978). 0.1 ml of culture medium for each of the four strains to be tested was spread on a sterilized agar medium, and the mixed extracts (M1 to M6) prepared in Preparation Example 1 were dissolved in the same solvent as the extraction solvent at a concentration of 10 mg/ml. After preparing the extraction sample, 10 μl or 20 μl was dropped onto a paper disk (Advantac, USA) with a diameter of 0.8 mm and cultured at 37°C. Afterwards, the antibacterial activity was measured based on the presence or absence of growth-inhibiting ring formation and the diameter of the growth-inhibiting ring.

Purified water was used as a control, and the antibacterial activity of each extract was evaluated and shown in [Table 2] below. For objective comparison, the results of the control group were fixed at an index of 10, and the results of the control group and the results of each extract were compared and evaluated on an index of 1 to 10. As for the index, the lower the number, the better the antibacterial activity.

con A.E. BE C.E. DE EE F.E. SA 10 4 6 6 4 6 5 EC 10 2 5 5 5 7 7 PA 10 4 6 5 6 6 7 MRSA 10 4 7 6 7 6 6

(Unit: index)

Referring to Table 2 above, it can be seen that AE has excellent antibacterial activity. Therefore, it can be confirmed that when using a single extract, when using AE, it can show the best antibacterial effect compared to the same dose.

In addition, in order to determine whether the use of the mixed formulation resulted in an additional synergistic effect on antibacterial activity, M1 to M12 were evaluated in the same manner as above. In addition, for an objective comparative evaluation of the synergistic effect, the result of AE as a control group was fixed at 10, and the antibacterial activity of M1 to M12 was comparatively evaluated with an index of 1 to 10. The lower the number, the better the antibacterial activity. If the index is 10, it means that the antibacterial activity is equal to or lower than that of AE.

The results are shown in [Table 3] below.

A.E. M1 M2 M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 SA 10 10 10 9 6 5 6 8 5 2 2 One 7 EC 10 10 10 9 7 6 6 8 5 3 2 2 6 PA 10 10 10 10 8 7 7 9 6 2 3 2 8 MRSA 10 10 10 10 8 8 7 9 6 3 2 One 7

Referring to [Table 3], it was confirmed that M1 to M2 had an overall low level of antibacterial activity compared to AE, and that a synergistic effect due to mixing was not observed.

However, in the case of the mixing range of M4 to M6, it can be confirmed that the antibacterial effect increases significantly due to the synergistic effect due to the interaction of the active ingredients included in each mixing composition. Therefore, it can be confirmed that excellent antibacterial activity can be achieved with a smaller amount using the above mixed composition. In addition, it can be seen that in the case of M9 to M11, additional synergistic activity appears compared to M5. This means that the above mixed formulations exhibit antibacterial activity through different mechanisms, and it can be assumed that a synergistic effect is achieved through interaction within a certain mixing range. Therefore, when within the above range, even a small amount can exhibit excellent antibacterial activity, making it possible to utilize it in various formulations.

2. Evaluation of anti-inflammatory activity

In order to evaluate the anti-inflammatory activity of each extract and mixed composition, hydrogen peroxide scavenging activity and DPPH free radical scavenging activity were evaluated.

Specifically, in the hydrogen peroxide scavenging ability test, hydrogen peroxide solution (40 mM) was prepared in phosphate buffer (pH 7.4). Black kelp extract was added to hydrogen peroxide solution (0.6mL, 40mM). The absorbance of hydrogen peroxide at 230 nm was measured after 10 minutes against a blank solution containing phosphate buffer without hydrogen peroxide. The percent hydrogen peroxide scavenging capacity of both sample and standard compounds was calculated:

[Equation 1]

Eliminated [H ₂ O ₂ ] = [(AC - AS)/AC] × 100

where AC is the absorbance of the control and AS is the absorbance in the presence of the sample or standard sample. The results are shown in [Table 4] below.

In addition, to evaluate the DPPH free radical scavenging ability, 10 mg DPPH was dissolved in 100 ml methanol (MeOH) to make it 100 ㎍/ml. A stock solution of the extract was prepared by dissolving 25 mg in 50 ml of MeOH. 1 ml DPPH was added to each solution and the solutions were kept dark and allowed to stand for exactly 30 minutes. The UV absorption of each solution was recorded at 516 nm. The reaction was carried out at 37 °C for 30 min and absorbance was monitored with a microplate reader at 516 nm. % radical scavenging activity (% RSA) was determined compared to the DMSO containing control. Scavenging of free radicals by DPPH as radical scavenging activity (%) was calculated as follows:

[Equation 1]

% RSA = (Control Absorbance - Absorbance)

The results are shown in [Table 4] below.

A.E. C.E. M1 M2 M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 H ₂ O ₂ scavenging activity (%) 64 78 81 75 66 58 52 53 73 59 42 38 44 62 DPPH scavenging activity (%) 86 90 91 88 85 74 68 68 88 77 58 53 58 81

Referring to [Table 4], excellent anti-inflammatory activity was observed in AE among single compositions. In particular, it can be seen that it has superior activity compared to CE.

In addition, in the mixed formulation, it can be seen that the anti-inflammatory activity increases due to the interaction of each active ingredient in M4 to M6, and in the case of M9 to M11, it shows a much better effect than the simple combination due to additional synergistic activity compared to M5. You can check the point.

Therefore, in the case of the above mixed formulation, it may be possible to use it in various formulations as a functional product that shows better effects with a smaller amount.

Therefore, the AE or mixed formulation provided in the present invention can exhibit excellent antibacterial and anti-inflammatory activity, and can be provided in various formulations such as food and external preparations. In particular, in the case of the above mixed formulation, excellent effects can be achieved with a small amount, so it can be provided as a functional product that can be used for a long time without problems such as side effects.

3. Evaluation of anti-allergic activity

In order to evaluate the resistance activity of allergic diseases for the AE and each mixed formulation, the production amount of prostaglandin D ₂ (PGD ₂ ) was measured. For measurements, the PGD ₂ EIA kit (Cayman, PGD2-MOX EIA kit, product no. 512011) was used.

Mouse bone marrow-derived mast cells (BMMC) were isolated from the bone marrow of BALB/C mice and cultured in RPMI-1640 medium containing 10% FBS, 100 U/ml penicillin, and 100 ng/ml streptomycin and IL. BMMCs of over 90% homogeneity were obtained by culturing with -3 (a culture medium containing the final 20% supernatant obtained by stimulating rat spleen cells with pokeweed mitogen) for approximately 3 weeks. After 3 ^weeks of culturing the BMMC above, the cells were pre-treated with AE at different concentrations (50 μg/ml, 100 μg/ml, 150 μg/ml, 200 μg/ml, 250 μg/ml) at a cell concentration of 2×10 5 cells for 30 minutes. , PGD ₂ in the supernatant 8 hours after cell stimulation was measured by EIA (Enzyme linked immune assay) using a PGD ₂ analysis kit (Cayman). At this time, in order to inhibit the production of PGD ₂ produced by COX-1 (cyclooxygenase-1), BMMCs were pretreated with 10 ng/ml of aspirin 1 hour before use. Washed four times with washing buffer, treated with 200 μl of substrate solution each, reacted for 5-20 minutes, treated with 50 μl of stop solution, and measured absorbance at 450 nm using ELx800. (BIO-TEK, Instrument, Inc, USA).

The results are shown in [Table 5] below.

50 100 150 200 250 PGD ₂ (pg/ml) 255 155 120 80 70

Referring to [Table 5], it can be seen that when AE is used, the production of PGD ₂ is suppressed in a concentration-dependent manner.

Meanwhile, in order to evaluate the synergistic activity of the mixed formulation, the same anti-allergy evaluation was conducted on M3 to M12, excluding M1 and M2, for which no synergistic effects were observed in antibacterial and anti-inflammatory effects. For objective comparative evaluation, the results of AE were fixed at an index of 10, and the results of M3 to M12 were compared with AE and evaluated with an index of 1 to 10. The lower the index number, the better the anti-allergy effect. Additionally, if the index is 10, it means that the effect is equal to or lower than that of AE and no synergistic effect is observed.

The results are shown in [Table 6] below.

A.E. M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 PGD ₂ (index) 10 10 7 7 6 9 7 5 4 5 8

Referring to [Table 6], it can be seen that in the case of M4 to M6, the anti-allergy effect increases due to additional synergistic activity. In addition, it can be confirmed that in the case of M9 to M11, an additional synergistic effect can be exhibited compared to M5.

Therefore, in the case of the above mixed formulation, it is possible to show a prevention and improvement effect on allergic diseases with a small amount through synergistic activity. In addition, it can be used continuously without problems such as side effects even after long-term use. In particular, in the case of representative allergic diseases such as asthma, eczema, and rhinitis, it is important to prevent and improve them through continuous use, so the mixed formulation according to the present invention can exhibit significantly excellent prevention and improvement effects.

Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims are also possible. falls within the scope of rights.

Claims (7)

Contains perilla perilla extract
Antibacterial and anti-inflammatory composition. According to clause 1,
The extract is extracted using an extraction solvent selected from the group consisting of water, alcohols with 1 to 6 carbon atoms, and mixtures thereof.
Antibacterial and anti-inflammatory composition. According to clause 2,
Gujeolcho extract; tangerine peel extract; Further comprising any one selected from the group consisting of Dalgae extract and mixtures thereof.
Antibacterial and anti-inflammatory composition. Comprising the composition according to claim 1
Functional food for antibacterial and anti-inflammatory purposes. Comprising the composition according to claim 1
Antibacterial and anti-inflammatory topical skin preparation. Containing the extract according to paragraph 1 or 2
Composition for preventing or improving allergic diseases. Containing the extract according to paragraph 1 or 2
Composition for preventing or improving asthma, eczema and rhinitis.