KR20220104929A - Composition for anti-inflammatory comprising a agastache rugosa extract as an active ingredient - Google Patents

Abstract

The present invention relates to an anti-inflammatory composition containing an Agastache rugose extract. The Agastache rugose extract is confirmed to effectively inhibit NO generated in an inflammatory reaction, so that the present invention provides a pharmaceutical composition and a health functional food composition for preventing or treating inflammatory diseases containing the Agastache rugose extract using the same.

Description

Anti-inflammatory composition comprising baechohyang extract as an active ingredient {COMPOSITION FOR ANTI-INFLAMMATORY COMPRISING A AGASTACHE RUGOSA EXTRACT AS AN ACTIVE INGREDIENT}

The present invention relates to a pharmaceutical composition for the prevention or treatment of inflammatory diseases comprising a baechohyang extract, and it has been confirmed that the baechohyang extract can effectively inhibit NO generated by an inflammatory reaction, and the inflammatory disease including the baechohyang extract using the same. A pharmaceutical composition for prevention or treatment is provided.

Inflammation occurs when tissues (cells) are damaged or infected with external infectious agents (bacteria, fungi, viruses, and various types of allergens), various inflammatory mediators and immune cells are involved to activate enzymes and mediate inflammation. It exhibits complex physiological reactions such as substance secretion, body fluid infiltration, cell migration, and tissue destruction and external symptoms such as erythema, edema, fever, and pain. In a normal case, the inflammatory reaction removes external infectious agents and regenerates damaged tissues, thereby restoring the function of the living organism. , in some cases rheumatoid arthritis, osteoporosis, sepsis, vascular disease, and cancer. That is, the inflammatory response is the most important response among the pathological mechanisms according to the host's defense mechanism against wounds and microbial infection, but the continuous and excessive inflammatory response damages the tissue.

Macrophages are the most representative immune cells regulating this inflammatory response, play an important role in the host's defense mechanism in a harmful environment, and are involved in the process of various diseases such as autoimmune diseases. Inflammatory factors such as lipopolysaccharide can activate macrophages, and the activated macrophages are tumor necrosis factor-α (TNF-α), interleukin-6 (interleukin-6, IL-6) and IL-1β, and synthesizes inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX2) to synthesize nitric oxide (NO ) and PGE 2 (prostaglandin E2).

On the other hand, Lipopolysaccharide (LPS) is a component of the outer membrane of Gram-negative bacteria and is known as an inflammatory factor. H2O2) and various cytotoxic substances such as nitric oxide function as an immune mechanism.

Recently, it has been reported that food or plants have the ability to prevent or treat diseases, and in response to the needs of mankind for a healthier and longer life, research to search for substances with various physiological functions from various resources around the world It is being actively carried out, and among them, a lot of attention is focused on compounds contained in plant resources. Therefore, the present inventors confirmed that the baechohyang extract can be used as a composition for the prevention or treatment of inflammatory diseases by confirming that it effectively inhibits NO, which is an inflammatory reaction, as a result of treating mouse-derived macrophages, and completed the present invention. .

An object of the present invention is to provide an anti-inflammatory composition comprising a baechohyang extract as an active ingredient, a pharmaceutical composition comprising the same, and a health functional food composition.

However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.

Hereinafter, the present invention will be described in detail. Advantages and features of the present invention, as well as embodiments for achieving them, will become apparent with reference to the following embodiments. However, the present invention is not limited to the embodiments published below, but can be implemented in various different forms, and only these embodiments make the publication of the present invention complete, and common knowledge in the technical field to which the present invention pertains It is provided to fully inform the possessor of the scope of the invention, and the present invention is only defined by the scope of the claims. Like reference numerals refer to like elements throughout.

Unless otherwise defined, all terms (including technical and scientific terms) used herein may be used with the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs. In addition, terms defined in a commonly used dictionary are not to be interpreted ideally or excessively unless clearly defined in particular. The terminology used herein is for the purpose of describing the embodiments and is not intended to limit the present invention. In this specification, the singular also includes the plural, unless specifically stated otherwise in the phrase.

In order to achieve the above object, the present invention provides an anti-inflammatory composition comprising an extract of baechohyang ( Agastache rugosa ) as an active ingredient.

The baechohyang extract may be a dried baechohyang extract extracted with water, C1 to C6 lower alcohol or a mixture thereof.

The concentration of the baechohyang extract may be 5 μM to 100 μM.

The baechohyang extract may inhibit inflammation induced in microglia activated by lipopolysaccharide (LPS).

The baechohyang extract can suppress the production of nitric oxide (NO) by the immune response.

The present invention also provides a pharmaceutical composition for preventing or treating inflammatory diseases, including an anti-inflammatory composition, and a health functional food composition for preventing or improving inflammatory diseases.

The inflammatory diseases include neuroinflammatory diseases, dermatitis, allergy, atopic dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatitis, lupus, fibromyalgia (fibromyalgia), psoriatic arthritis, osteoarthritis, rheumatoid arthritis, paro-scapularis, tendinitis, tendinitis, peritenitis, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, acute and chronic inflammatory diseases. have.

The present invention relates to a composition for the prevention, treatment or improvement of inflammatory diseases comprising a baechohyang extract as an active ingredient, wherein the baechohyang extract inhibits inflammation induced in microglia activated by lipopolysaccharide (LPS). , it is excellent in the prevention, treatment or improvement of inflammatory diseases, so it can be usefully used as a pharmaceutical or health functional food.

1 is a result confirming the effect of the above-ground part methanol extract of baechohyang in LPS-stimulated BV-2 microglia on cell viability and the effect of reducing the amount of nitrogen monoxide released.

Hereinafter, to help the understanding of the present invention, examples will be described in detail. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

The present inventors have confirmed that when the methanol extract of baechohyang is treated to BV-2 microglia activated by lipopolysaccharide (LPS), it can inhibit inflammation in a concentration-dependent manner while having a high cell viability. The invention was completed.

Pharmaceutical composition for preventing or treating inflammatory diseases

The present invention provides an anti-inflammatory composition comprising baechohyang extract as an active ingredient and a pharmaceutical composition for preventing or treating inflammatory diseases comprising the same.

The pharmaceutical composition for preventing or treating inflammatory diseases according to the present invention comprises a baechohyang extract as an active ingredient.

The baechohyang (Agastache rugosa) is a dicotyledonous plant, a perennial herb of the Lamiaceae family, and is also called Guanghyang (藿香) in China. Kwakhyang is known to be effective in digestion, stomach pain, pain, vomiting, stomach pain, colds, etc.

The baechohyang extract may be one obtained by extracting baechohyang with water, C1 to C6 lower alcohol or a mixture thereof (especially, ethanol).

Specifically, the extraction is preferably performed by adding 2 to 20 times the volume of ginger, and 3 to 15 times the volume of water, C1 to C4 lower alcohol or a mixture thereof after shredding dried baecho flavor. It is more preferable to extract, but is not limited thereto. The extraction temperature is preferably 30°C to 100°C, more preferably 30°C to 70°C, but is not limited thereto. The extraction time is preferably 1 hour to 48 hours, more preferably 12 hours to 36 hours, but is not limited thereto. As the extraction method, cold extraction, ultrasonic extraction, or reflux cooling extraction method are all available, but the present invention is not limited thereto. The number of extractions is preferably 1 to 5 times, and more preferably 2 to 3 times repeated extraction, but is not limited thereto. Additionally, the baechohyang extract may be diluted, concentrated, or purified and used after dilution or concentration.

The baechohyang extract inhibits inflammation induced in microglia activated by lipopolysaccharide (LPS), and specifically, the inhibition of the inflammation can be confirmed by inhibiting the production of nitric oxide (NO).

In addition, the concentration of the baechohyang extract is preferably 5 μM to 100 μM, but is not limited thereto, and preferably 5 μM to 25 μM. At this time, when the concentration of the baechohyang extract is too low, since the inflammation induced in microglia activated by lipopolysaccharide (LPS) cannot be effectively suppressed, the prevention, treatment or improvement effect of inflammatory diseases is insignificant. If there is, and the concentration of the baechohyang extract is too high, there may be concerns of toxicity, including cytotoxicity.

The inflammatory disease is an inflammatory mediator induced in microglia activated by lipopolysaccharide (LPS), neuroinflammatory disease, dermatitis, allergy, atopic dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, pharyngitis, tonsillitis, pneumonia, Gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatoid arthritis, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, periarthritis, tendinitis, tendinitis, periostitis, myositis, hepatitis, cystitis, It is preferably selected from the group consisting of nephritis, sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases, but is not limited thereto.

The pharmaceutical composition for preventing or treating inflammatory diseases according to the present invention is in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups and aerosols, external preparations, suppositories, and sterile injection solutions, respectively, according to conventional methods. It may be formulated and used, and may include an appropriate carrier, excipient or diluent commonly used in the preparation of a pharmaceutical composition for formulation.

The carrier or excipient or diluent may include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, undecided. various compounds or mixtures including vaginal cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

In the case of formulation, it can be prepared using commonly used diluents or excipients such as fillers, weight agents, binders, wetting agents, disintegrants, and surfactants.

A solid preparation for oral administration may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like with the baechohyang extract. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used.

Liquid formulations for oral use include suspensions, solutions, emulsions, syrups, etc. In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included. .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous preparations, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As a base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol gelatin and the like can be used.

The preferred dosage of the pharmaceutical composition for preventing or treating inflammatory diseases according to the present invention varies depending on the patient's condition, body weight, degree of disease, drug form, administration route and period, but may be appropriately selected by those skilled in the art. However, for a desirable effect, it may be administered at 0.0001 to 2,000 mg/kg per day, preferably 0.001 to 2,000 mg/kg. Administration may be administered once a day, or may be administered in several divided doses. However, the scope of the present invention is not limited by the dosage.

The pharmaceutical composition for preventing or treating inflammatory diseases according to the present invention may be administered to mammals such as mice, mice, livestock, and humans by various routes. All modes of administration can be administered by, for example, oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

Health functional food for preventing or improving inflammatory diseases

In addition, the present invention provides a health functional food for preventing or improving inflammatory diseases comprising a baechohyang extract as an active ingredient.

The health functional food for preventing or improving inflammatory diseases according to the present invention includes the baechohyang extract as an active ingredient, and since the baechohyang extract has been described above, the redundant description will be omitted.

The inflammatory disease is an inflammatory mediator induced in microglia activated by lipopolysaccharide (LPS), neuroinflammatory disease, dermatitis, allergy, atopy, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia , gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatoid arthritis, lupus, fibromyalgia, psoriatic arthritis, osteoarthritis, rheumatoid arthritis, parietal sinusitis, tendinitis, tendinitis, periostitis, myositis, hepatitis, cystitis , nephritis, Sjogren's syndrome, multiple sclerosis, and acute and chronic inflammatory diseases are preferably selected from the group consisting of, but not limited to.

In the health functional food for preventing or improving inflammatory disease according to the present invention, when the baechohyang extract is used as an additive for health functional food, it can be added as it is or used with other food or food ingredients, and is suitable according to a conventional method. can be used The mixing amount of the active ingredient may be appropriately determined according to each purpose of use, such as prevention, health or treatment.

The formulation of the health functional food may be in the form of powders, granules, pills, tablets, and capsules, as well as in the form of general food or beverages.

The type of food is not particularly limited, and examples of food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, and dairy products including ice cream. , various soups, beverages, teas, drinks, alcoholic beverages, and vitamin complexes, and may include all foods in a conventional sense.

In general, when preparing food or beverage, the baechohyang extract may be added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less, based on 100 parts by weight of the raw material. However, in the case of long-term intake for health and hygiene or health control, the amount may be less than the above range, and since the present invention does not have a problem in terms of safety in terms of using a natural product, the amount above the above range can also be used.

The beverage among the health functional foods according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, like conventional beverages. The above-mentioned natural carbohydrates may be monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatine and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate may be about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the beverage according to the present invention.

In addition to the above, the health functional food for preventing or improving inflammatory diseases according to the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusting agents, It may contain stabilizers, preservatives, glycerin, alcohols, and carbonating agents used in carbonated beverages. In addition, the health functional food for preventing or improving inflammatory diseases of the present invention may contain fruit for the production of natural fruit juice, fruit juice beverage, and vegetable beverage. These components may be used independently or in combination. The ratio of these additives is not limited, but is generally selected in the range of 0.01 to 0.1 parts by weight relative to 100 parts by weight of the health functional food of the present invention.

Accordingly, the present invention relates to a composition for preventing, treating or improving inflammatory diseases comprising a baechohyang extract as an active ingredient, wherein the baechohyang extract inhibits inflammation induced in microglia activated by lipopolysaccharide (LPS). By doing so, the prevention, treatment or improvement effect of inflammatory diseases is excellent, and thus it can be usefully used as a pharmaceutical or health functional food.

Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.

material preparation

Methanol extract from baechohyang, Lipopolysaccharide (LPS), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). (Sigma Chemical Co, USA) 24well tissue culture plate, 60mm culture plate, 100mm culture plate. (Falcon, IL, USA) For the reagent used in the experiment, DMEM was used as a medium for cell culture, and 100 units/ml penicillin/streptomycin (Gibco, BRL) was used as 5% FBS and antibiotics.

Cell culture

BV-2 microglial cells were cultured in DMEM media supplemented with inactivated 5% FBS and 100 units/ml 1% penicillin/streptomycin and 5% CO ₂ incubator. In all experiments, the number of cells was 2.5 × 10 ⁵ cell/ml. The treatment of the methanol extract of baechohyang was diluted in the medium and pre-treated for 1 hour at each concentration (5, 10 and 25 μg/ml), and 200 ng/ml of LPS was used.

The cytotoxicity test of the methanol extract of baechohyang can be confirmed by MTT assay. In the MTT assay, after NO assay, 20 μl of 5 mg/ml MTT is added to the cells in the microplate, and reacted in an incubator for 1 hour to incubate formazan crystals with 400 μl of DMSO. The absorbance was measured with a microplate reader at a wavelength of 540 nm.

Assay of NO

A method for quantitatively measuring the production of NO (nitric oxide) is a microplate (24 well) assay method. After seeding the cells in the microplate, LPS and baechohyang methanol extract were added at different concentrations and reacted in a 5% CO ₂ incubator. After 24 hours, 100 μl of the supernatant was reacted with Griess reagent (1% sulfanilamide, 0.1% N-(1-naphthyl)-ethylenediamine dihydrochloride, 2.5% H ₃ PO ₄ ) in a 96 well plate, and then microplate at 540 nm wavelength. Absorbance was measured with a reader.

Preparation of baechohyang extract

The baechohyang extract used in the experiment was extracted for 24 hours at room temperature with 1L of 100% ethanol, about 10 times the volume of 100g of dried baechohyang (purchased from Gyeongdong Market), and the filtered extract was vacuum-concentrated at 30~70℃ and freeze-dried, It was dissolved in DMSO and used for the experiment (yield is about 9.39%).

[Experimental Example 1]

1-1. Cytotoxicity test of baechohyang extract

First, toxicity test was performed on the methanol extract of baechohyang through MTT assay. Based on 100% of the cell viability of the control group not treated with anything, 25 μM, a negative control group treated with LPS and substances, 25 μM, 10 μM, and 25 μM, and a methanol extract of baechohyang alone were compared. 89.74 ± 4.82%, 96.03 ± 7.37%, 91.62 ± 4.92%, 79.83 ± 7.63%, and 94.32 ± 5.75%, respectively, were confirmed to have no toxicity to the methanol extract of baechohyang, and it was confirmed that there was no specific effect of the substance in the use of a negative control. was confirmed (Fig. 1A).

1-2. Measurement of the inhibitory effect of nitric oxide (NO) production in LPS-stimulated BV-2 microglia when treated with baechohyang methanol extract

In order to confirm the neuroinflammation inhibitory efficacy of the methanol extract of baechohyang, the inhibitory efficacy of nitric oxide (NO) in BV-2 microglia was confirmed through NO assay. Through this, the nitric oxide inhibitory efficacy of the methanol extract of baechohyang, a candidate for neuroinflammation, was confirmed in BV-2 microglia.

After BV-2 microglia were placed in a 24-well plate at a volume of 2.5 × 10 ⁵ cells/well, the methanol extract of Baechohyang was pretreated for 1 hour at each concentration. Then, the cells were stimulated with LPS (200 ng/ml). After 24 hours, 100 μl of the cell supernatant was dispensed into 96 wells and the amount of nitric oxide released was measured by Griess assay. After adding MTT reagent to the remaining supernatant, the value obtained by measuring the surviving cells was expressed as a percentage. (#, control), (**, P<0.001), (***, P<0.0001)

NO assay was performed at the same concentration as in Experimental Example 1-1. The value in the untreated group was 1.61 ± 0.1 μM, and the value in the group treated with LPS alone was 11.72 ± 0.84 μM. When each of the baechohyang methanol extracts were treated, 5 μM was 9.77 ± 0.79 μM, 10 μM was 5.83 ± 0.66 μM, and 25 μM was 0.72 ± 0.07 μM, confirming that the nitric oxide inhibitory effect was inhibited depending on the concentration of the baechohyang methanol extract. There was (Fig. 1B). Therefore, it is judged that the methanol extract of baechohyang will be effective in suppressing nitric oxide.

Claims (9)

An anti-inflammatory composition comprising an extract of baechohyang ( Agastache rugosa ) as an active ingredient. The method of claim 1,
The baechohyang extract is an anti-inflammatory composition that is extracted with water, C1 to C6 lower alcohol or a mixture thereof. The method of claim 1,
The concentration of the baechohyang extract is 5 μM to 100 μM, anti-inflammatory composition. The method of claim 1,
The baechohyang extract is an anti-inflammatory composition, characterized in that it inhibits inflammation induced in microglia activated by lipopolysaccharide (LPS). The method of claim 1,
The baechohyang extract is characterized in that it suppresses the production of nitric oxide (NO) by the immune response, anti-inflammatory composition. A pharmaceutical composition for preventing or treating inflammatory diseases, comprising the anti-inflammatory composition of claim 1. 7. The method of claim 6,
The inflammatory disease is a neuroinflammatory disease, dermatitis, allergy, atopic dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatitis, lupus, fibromyalgia group consisting of (fibromyalgia), psoriatic arthritis, osteoarthritis, rheumatoid arthritis, parotiditis, tendinitis, tendinitis, paronychia, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, acute and chronic inflammatory diseases At least one disease selected from, a pharmaceutical composition. A health functional food composition for preventing or improving inflammatory diseases, comprising the anti-inflammatory composition of claim 1. 9. The method of claim 8,
The inflammatory disease is a neuroinflammatory disease, dermatitis, allergy, atopic dermatitis, conjunctivitis, periodontitis, rhinitis, otitis media, sore throat, tonsillitis, pneumonia, gastric ulcer, gastritis, Crohn's disease, colitis, hemorrhoids, gout, ankylosing spondylitis, rheumatitis, lupus, fibromyalgia group consisting of (fibromyalgia), psoriatic arthritis, osteoarthritis, rheumatoid arthritis, parotiditis, tendinitis, tendinitis, paronychia, myositis, hepatitis, cystitis, nephritis, sjogren's syndrome, multiple sclerosis, acute and chronic inflammatory diseases At least any one disease selected from, health functional food composition.

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