Composition for pain suppression comprising perilla leaf extract and method for preparing same

Abstract

The present invention relates to: a pharmaceutical composition for pain suppression, the composition containing a perilla leaf extract as an active ingredient; and a method for preparing same. The pharmaceutical composition can be used as a health functional food and feed composition for pain suppression.

Description

Pain-relieving composition containing perilla leaf extract and method for producing the same

[0001] The present invention relates to a composition exhibiting pain-suppressing activity, and more specifically, to a pharmaceutical composition and a health functional food composition having pain-suppressing efficacy, which contain a perilla leaf extract as an active ingredient.

[0002]

[0003] Perilla seeds are emerging as a high-income crop, with demand rivaling sesame oil, as their previously unknown benefits—anti-anemia, vascular health, improved cholesterol levels, and boosted immunity—are gaining recognition. Perilla seeds are not only environmentally friendly and pest-free, allowing for pesticide-free cultivation, but also suffer minimal damage to crops. Furthermore, their relatively low cost and ease of cultivation in aging rural areas are contributing to increased farm income.

[0004] Perilla oil exports are currently diversifying beyond Japan to the US, Singapore, and Taiwan, fueling continued domestic production expansion. Perilla oil contains over 60% alpha-linolenic acid, an omega-3. In addition to its unique vegetable oil flavor, it boasts health benefits and is emerging as a globally exportable food ingredient. In 2021, perilla oil was approved as a functional food in Japan, and studies have shown that consuming 4.7 g per day is beneficial for high blood pressure.

[0005] Perilla and Purslane Extract (No. 2021-11) has been certified for its ability to help boost immune function. Despite the proven usefulness of perilla leaves in traditional Korean medicine over a long period of time, the pain-relieving activity of perilla leaves themselves has not been utilized in various compositions.

[0006] Accordingly, the inventors of the present invention conducted experiments on the pain-relieving effects of perilla leaf extract as an active ingredient and verified the effects with data. Through this, the inventors of the present invention seek to provide an invention relating to a pharmaceutical composition for pain relief containing perilla leaf extract as an active ingredient.

[0007]

[0008] The main purpose of the present invention is to provide a pharmaceutical composition and a health functional food composition showing pain suppressing efficacy containing perilla leaf extract as an active ingredient.

[0009] Another object of the present invention is to provide a method for inhibiting the expression of cytokines IL-1β, IL-6, GM-CSF and TNF-α, which comprises administering the pharmaceutical composition to a subject.

[0010]

[0011] The terminology used herein is for the purpose of description only and should not be construed as limiting. The singular expression includes the plural expression unless the context clearly indicates otherwise. In this specification, the terms "comprises" or "has" and the like are intended to specify the presence of a feature, number, step, operation, component, part, or combination thereof described in the specification, but should be understood to not preclude the possibility of the presence or addition of one or more other features, numbers, steps, operations, components, parts, or combinations thereof.

[0012] Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by a person of ordinary skill in the art to which the embodiments pertain. Terms defined in commonly used dictionaries should be interpreted as having a meaning consistent with their meaning in the context of the relevant technology, and shall not be interpreted in an idealized or overly formal sense unless explicitly defined herein.

[0013] The present invention provides a pharmaceutical composition for pain relief containing a perilla leaf extract as an active ingredient.

[0014] Preferably, the extract may be an ethanol extract, but is not limited thereto.

[0015] The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients, and diluents commonly used in the manufacture of pharmaceutical compositions. The excipients may be, for example, one or more selected from the group consisting of diluents, binders, disintegrants, lubricants, adsorbents, moisturizers, film-coating materials, and controlled-release additives.

[0016] The pharmaceutical composition according to the present invention may be formulated and used in the form of external preparations such as powders, granules, sustained-release granules, enteric-coated granules, liquids, eye drops, ellipsoids, emulsions, suspensions, alcohols, troches, aromatic waters, limonades, tablets, sustained-release tablets, enteric-coated tablets, sublingual tablets, hard capsules, soft capsules, sustained-release capsules, enteric capsules, pills, tinctures, soft extracts, dry extracts, fluid extracts, injections, capsules, irrigation solutions, ointments, lotions, pastes, sprays, inhalants, patches, sterile injection solutions, or aerosols, according to a conventional method, and the external preparations may have formulations such as creams, gels, patches, sprays, ointments, ointments, lotions, liniments, pastes, or cataplasmas.

[0017] Carriers, excipients and diluents that may be included in the pharmaceutical composition according to the present invention include lactose, dextrose, sucrose, oligosaccharides, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

[0018] When formulating, it is usually prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants.

[0019] The additives of the tablets, powders, granules, capsules, pills, and troches according to the present invention include excipients such as corn starch, potato starch, wheat starch, lactose, sucrose, glucose, fructose, D-mannitol, precipitated calcium carbonate, synthetic aluminum silicate, calcium hydrogen phosphate, calcium sulfate, sodium chloride, sodium bicarbonate, purified lanolin, microcrystalline cellulose, dextrin, sodium alginate, methylcellulose, sodium carboxymethylcellulose, kaolin, urea, colloidal silica gel, hydroxypropyl starch, hydroxypropyl methylcellulose (HPMC) 1928, HPMC 2208, HPMC 2906, HPMC 2910, propylene glycol, casein, calcium lactate, and Primogel; Gelatin, gum arabic, ethanol, agar powder, cellulose acetate phthalate, carboxymethylcellulose, calcium carboxymethylcellulose, glucose, purified water, sodium caseinate, glycerin, stearic acid, sodium carboxymethylcellulose, sodium methylcellulose, methylcellulose, microcrystalline cellulose, dextrin, hydroxycellulose, hydroxypropyl starch, hydroxymethylcellulose, refined shellac, starch starch, hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinyl alcohol, polyvinyl pyrrolidone, and binders such as hydroxypropyl methylcellulose, corn starch, agar powder, methylcellulose, bentonite, hydroxypropyl starch, sodium carboxymethylcellulose, sodium alginate, Disintegrants such as carboxymethylcellulose calcium, calcium citrate, sodium lauryl sulfate, anhydrous silicic acid, 1-hydroxypropyl cellulose, dextran, ion exchange resin, polyvinyl acetate, formaldehyde-treated casein and gelatin, alginic acid, amylose, guar gum, baking soda, polyvinylpyrrolidone, calcium phosphate, gelling starch, gum arabic, amylopectin, pectin, sodium polyphosphate, ethyl cellulose, sucrose, magnesium aluminum silicate, di-sorbitol solution, and light anhydrous silicic acid;Lubricants such as calcium stearate, magnesium stearate, stearic acid, hydrogenated vegetable oil, talc, lycopodium dentata, kaolin, petrolatum, sodium stearate, cacao butter, sodium salicylate, magnesium salicylate, polyethylene glycol 4000, polyethylene glycol 6000, liquid paraffin, hydrogenated soybean oil (Lubri wax), aluminum stearate, zinc stearate, sodium lauryl sulfate, magnesium oxide, macrogol, synthetic aluminum silicate, anhydrous silicic acid, higher fatty acids, higher alcohols, silicone oil, paraffin oil, polyethylene glycol fatty acid ether, starch, sodium chloride, sodium acetate, sodium oleate, dl-leucine, and light anhydrous silicic acid can be used.

[0020] As additives of the liquid formulation according to the present invention, water, dilute hydrochloric acid, dilute sulfuric acid, sodium citrate, monostearate sucrose, polyoxyethylene sorbitol fatty acid esters (twin esters), polyoxyethylene monoalkyl ethers, lanolin ethers, lanolin esters, acetic acid, hydrochloric acid, ammonia water, ammonium carbonate, potassium hydroxide, sodium hydroxide, prolamine, polyvinylpyrrolidone, ethyl cellulose, sodium carboxymethyl cellulose, etc. can be used.

[0021] The syrup according to the present invention may include a solution of white sugar, other sugars, or sweeteners, and may also include a fragrance, a coloring agent, a preservative, a stabilizer, a suspending agent, an emulsifier, a viscosity modifier, and the like, as needed.

[0022] Purified water may be used in the emulsion according to the present invention, and emulsifiers, preservatives, stabilizers, fragrances, etc. may be used as needed.

[0023] The suspension according to the present invention may include suspending agents such as acacia, tragacanth, methylcellulose, carboxymethylcellulose, sodium carboxymethylcellulose, microcrystalline cellulose, sodium alginate, hydroxypropylmethylcellulose (HPMC), HPMC 1828, HPMC 2906, and HPMC 2910, and, if necessary, surfactants, preservatives, stabilizers, colorants, and fragrances may be used.

[0024] The injection according to the present invention includes: a solvent such as distilled water for injection, 0.9% sodium chloride injection, Ringer's injection, dextrose injection, dextrose + sodium chloride injection, PEG, lactated Ringer's injection, ethanol, propylene glycol, non-volatile oils - sesame oil, cottonseed oil, peanut oil, soybean oil, corn oil, ethyl oleate, isopropyl myristate, and benzene benzoate; a solubilizing agent such as sodium benzoate, sodium salicylate, sodium acetate, urea, urethane, monoethylacetamide, butazolidine, propylene glycol, tween, nitrile acid amide, hexamine, and dimethylacetamide; a buffer such as weak acids and their salts (acetic acid and sodium acetate), weak bases and their salts (ammonia and ammonium acetate), organic compounds, proteins, albumin, peptone, and gums; an isotonic agent such as sodium chloride; It may contain stabilizers such as sodium bisulfite (NaHSO3), carbon dioxide gas, sodium metabisulfite (Na2S2O5), sodium sulfite (Na2SO3), nitrogen gas (N2), and ethylenediaminetetraacetic acid; oxidizing agents such as sodium bisulfite 0.1%, sodium formaldehyde sulfoxylate, thiourea, disodium ethylenediaminetetraacetic acid, and acetone sodium bisulfite; analgesics such as benzyl alcohol, chlorobutanol, procaine hydrochloride, glucose, and calcium gluconate; and suspending agents such as sodium citric acid, sodium alginate, Tween 80, and aluminum monostearate.

[0025] The suppository according to the present invention comprises cocoa butter, lanolin, withepsol, polyethylene glycol, glycerogelatin, methylcellulose, carboxymethylcellulose, a mixture of stearic acid and oleic acid, Subanal, cottonseed oil, peanut oil, palm oil, cocoa butter + cholesterol, lecithin, ranet wax, glycerol monostearate, Tween or Span, Imhausen, monolene (propylene glycol monostearate), glycerin, Adeps solidus, Buytyrum Tego-G, Cebes Pharma 16, hexalide base 95, Cotomar, Hydroxocote SP, S-70-XXA, S-70-XX75 (S-70-XX95), Mechanisms such as Hydrokote 25, Hydrokote 711, Idropostal, Massa estrarium (A, AS, B, C, D, E, I, T), Massa-MF, Masupol, Masupol-15, Neosupostal-N, Paramound-B, Suposiro (OSI, OSIX, A, B, C, D, H, L), Suppository type IV (AB, B, A, BC, BBG, E, BGF, C, D, 299), Supostal (N, Es), Wecovi (W, R, S, M, Fs), and Tezester triglyceride basis (TG-95, MA, 57) can be used.

[0026] Solid dosage forms for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid dosage forms are prepared by mixing the extract with at least one excipient, such as starch, calcium carbonate, sucrose or lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used.

[0027] Liquid preparations for oral administration include suspensions, solutions, emulsions, and syrups. In addition to commonly used simple diluents such as water and liquid paraffin, they may contain various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.

[0028] The pharmaceutical composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease with a reasonable benefit / risk ratio applicable to medical treatment. The effective dosage level can be determined based on factors including the type and severity of the patient's disease, drug activity, drug sensitivity, administration time, administration route and excretion rate, treatment period, concurrently used drugs, and other factors well known in the medical field.

[0029] The pharmaceutical composition according to the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents. It can be administered sequentially or simultaneously with conventional therapeutic agents, or in single or multiple doses. Taking all of the above factors into account, it is important to administer an amount that achieves maximum efficacy with minimal side effects. This amount can be readily determined by those skilled in the art to which the present invention pertains.

[0030] The pharmaceutical composition of the present invention can be administered to a subject via various routes. All modes of administration are conceivable, including oral ingestion, subcutaneous injection, intraperitoneal administration, intravenous injection, intramuscular injection, intrathecal injection, sublingual administration, buccal mucosal administration, rectal insertion, vaginal insertion, ocular administration, otic administration, nasal administration, inhalation, oral or nasal spraying, dermal administration, and transdermal administration.

[0031] The pharmaceutical composition of the present invention is determined according to the type of drug as an active ingredient along with various related factors such as the disease to be treated, route of administration, age, sex, weight of the patient, and severity of the disease.

[0032] In the present invention, the term "subject" means a subject requiring treatment for a disease, and more specifically, a mammal such as a human or non-human primate, mouse, rat, dog, cat, horse, and cow.

[0033] In the present invention, "administration" means providing a predetermined composition of the present invention to a subject by any suitable method. In the present invention, "prevention" means any action that inhibits or delays the onset of a target disease, "treatment" means any action that improves or beneficially changes a target disease and its resulting metabolic abnormality symptoms by administering a pharmaceutical composition according to the present invention, and "improvement" means any action that reduces a parameter related to a target disease, for example, the severity of a symptom, by administering a composition according to the present invention.

[0034] In the present invention, the perilla leaf extract may exhibit an effect through a mechanism of inhibiting NO production or cytokine expression, and the type of cytokine may preferably be at least one selected from the group consisting of TNF-α, IL-6, IL-1β, or GM-CSF, but is not limited thereto.

[0035] The present invention also provides a health functional food composition for pain relief containing the perilla leaf extract.

[0036] In the present invention, "food" means a natural product or processed product containing one or more nutrients, preferably a product that has gone through a certain degree of processing to become directly edible, and in its usual sense, includes all health functional foods, beverages, food additives, and beverage additives.

[0037] In the present invention, the term "functional food" is the same as food for special health use (FoSHU), and refers to a food with high medical or healthcare effects that is processed to efficiently exhibit a bioregulatory function in addition to providing nutrition, and may be manufactured in the form of tablets, capsules, pills, granules, powders, liquids, flakes, pastes, syrups, gels, jellies, bars, or films. Here, "function" means regulating nutrients for the structure and functions of the human body or obtaining a useful effect for health purposes such as physiological actions.

[0038] When the perilla leaf extract of the present invention is used as a food additive, it can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The mixing amount of the active ingredient can be appropriately determined depending on the purpose of use (prevention, health, or therapeutic treatment). Generally, when manufacturing a food or beverage, the perilla leaf extract of the present invention can be added in an amount of 15% by weight or less, or 10% by weight or less, based on the raw material. However, in the case of long-term intake for the purpose of health and hygiene or health control, the amount can be below the above range, and since there is no problem in terms of safety, the active ingredient can also be used in an amount above the above range.

[0039] There are no specific restrictions on the types of the above foods. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and all health functional foods in the conventional sense are included.

[0040] The health beverage composition according to the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients, similar to conventional beverages. The natural carbohydrates mentioned above include 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 a sweetener, natural sweeteners such as thaumatin and stevia extract, or synthetic sweeteners such as saccharin and aspartame may be used. The proportion of the natural carbohydrate is generally about 0.01-0.20 g, or about 0.04-0.10 g, per 100 mL of the composition of the present invention.

[0041] In addition to the above, the composition of the present invention may contain various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and its salts, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, etc. In addition, the composition of the present invention may contain fruit pulp for the production of natural fruit juice, fruit juice drinks, and vegetable drinks. These ingredients may be used independently or in combination. The proportion of these additives is not particularly critical, but is typically selected within the range of 0.01 to 0.20 parts by weight per 100 parts by weight of the composition of the present invention.

[0042] The present invention also provides a feed composition for suppressing pain comprising the perilla leaf extract.

[0043] The term "feed" in the present invention means any natural or artificial diet, meal, etc. or a component of said meal for eating, ingesting, and digesting by an animal or suitable therefor.

[0044] The type of the above feed is not particularly limited, and feed commonly used in the relevant technical field can be used. Non-limiting examples of the above feed include plant-based feed such as grains, roots, food processing by-products, algae, fiber, pharmaceutical by-products, oils, starches, meal, or grain by-products; and animal-based feed such as proteins, inorganic substances, oils, minerals, oils, single-cell proteins, zooplankton, or food. These may be used alone or in combination of two or more.

[0045]

[0046] The perilla leaf extract according to the present invention exhibits pain-suppressing efficacy, and can be used as a pharmaceutical composition for pain suppression and a health functional food composition containing the extract as an active ingredient.

[0047]

[0048] Figure 1 is a diagram showing the inhibition rate of NO production and cytokine expression of perilla leaf extract.

[0049] Figure 2 is a diagram showing the pain-suppressing and swelling-reducing effects of perilla leaf extract in an animal model.

[0050]

[0051] Hereinafter, preferred examples are presented to aid in understanding the present invention. However, the following examples are provided solely to facilitate a better understanding of the present invention, and the scope of the present invention is not limited by the following examples.

[0052]

[0053] Example 1. Preparation of perilla leaf extract

[0054] After washing the raw material, the perilla leaves were blanched in hot water at 100℃ for 1-2 minutes and then dried in low-temperature hot air at 45℃ for 5 days. The leaves were then selected, the stems removed, and ground. 1,900 g of 70% ethanol was used as a solvent to extract 100 g of dried perilla leaves by stirring at 100 rpm for 24 hours at room temperature. The extract was filtered through a 5-8 μm filter, concentrated to about 80%, centrifuged at 11,000 rpm for 30 minutes, the supernatant was discarded, suspended in 400 g of distilled water, and vortexed. After that, the leaves were sonicated at 65℃ for 30 minutes, filtered through a 5-8 μm filter, and dried in hot air at 82℃ for 3 days before being finely ground.

[0055]

[0056] Example 2. Inhibitory effects of perilla leaf extract on NO production and cytokine expression

[0057] When macrophages are induced by external stimuli such as LPS, they secrete NO, produce various substances such as inflammatory cytokines, and initiate various pathological responses that regulate the inflammatory response. The composition was treated in RAW264.7 cells induced by LPS to investigate its efficacy.

[0058] The stabilized NO oxide, NO2 (Nitrite), was measured using the Griess reaction. First, mouse macrophage cell line (RAW264.7) was seeded in a 96-well plate at 5 x 10 per well. 4After dispensing cells / 100μl, CO2 culture was performed for 24 hours. LPS (lipopolysaccharide) was treated at a concentration of 1μg / ml and CO2 cultured for 1 hour, then the prepared perilla leaf extract was treated and CO2 cultured for 24 hours. Then, each culture supernatant was placed in a 96-well plate, and an equal volume of Griess reagent (0.1% N-1-naphtyl-ethylendiamine in H2O: 1% sulfanilamidein 5% H3PO4 = 1:1) was added and reacted for 10 minutes, and the absorbance was measured at 550 nm using a microplate reader. The concentration of NO2 (Nitrite) was calculated by comparing it with a standard curve obtained by diluting NaNO2 (sodium nitrite) 2-fold from 100μM to 0.7μM. LPS was used as a positive control.

[0059] As a result, it was confirmed that the production of NO was suppressed by approximately 50% in the perilla leaf extract treatment group compared to the control group (Figure 1A).

[0060] RAW264.7 macrophages were seeded at 5 x 10 per well in a 24-well plate. 5Cells were dispensed at 1 ml / well and cultured in CO2 for 24 hours. Afterwards, LPS (lipopolysaccharide) was treated at a concentration of 1 μg / ml in the well and cultured in CO2 for 24 hours. Then, the cell culture supernatant was collected. Cytokines IL-1β, IL-6, GM-CSF, and TNF-α contained in the supernatant were measured using enzyme-linked immunosorbent assay (ELISA). That is, the primary antibody (capture antibody) was diluted in coating solution (0.1 M sodium carbonate, pH 9.5) and dispensed at 100 μl / well into plate-bottom micro-wells, cultured overnight at 4°C, and then washed with washing solution (0.05% Tween 20 / PBS). The washed microwells were blocked with PBS containing 10% FBS, and the culture supernatant collected in the experiment was diluted to an appropriate ratio and dispensed into each well and reacted at room temperature. Next, 100 μl / well of biotinylated secondary antibody was reacted at room temperature for a certain period of time, and then 100 μl / well of avidin-peroxidase (enzyme reagent) was added. Finally, the substrate (3, 3′, 5, 5′-Tetramethylbenzidine Liquid Substrate, H2O2) was added to develop the color, and then measured using a microplate reader. The measured concentrations of IL-1β, IL-6, GM-CSF, and TNF-α were converted using a standard curve.

[0061] As a result, it was confirmed that the expression of inflammatory cytokines IL-1β (Fig. 1B), IL-6 (Fig. 1C), TNF-α (Fig. 1E), and GM-CSF (Fig. 1D), a cytokine that functions as a growth factor for leukocytes, was significantly reduced in the perilla leaf extract treatment group compared to the control group.

[0062]

[0063] Example 3. Pain-inhibiting efficacy of perilla leaf extract in an animal model

[0064] Seven-week-old Sprague-Dawley (SD) rats were housed in cages and fed ad libitum solid rodent diet (Teklad certified irradiated global 18% protein rodent diet, 2918C, Inotiv, USA) and tap water sterilized with UV sterilizer and microfiltration. After acclimatization for approximately 7 days, the general health status was observed, and healthy rats were selected for use in the experiment. The laboratory environment was a temperature of 22°C ± 3°C, a humidity of 55% ± 15%, and a 12-h light-dark cycle (lights on: 8:00 AM).

[0065] Rats acclimated to the breeding room for 7 days were subjected to inhalation anesthesia, and 50 μl of CFA was injected between the tibiofibular and tarsus bones. Afterwards, the degree of pain and swelling was measured using a Vonfrey filament, and daily body weight was measured to monitor for abnormalities in food intake. Mechanical allodynia was measured using Dixon's up-down method.

[0066] This study was approved by the Animal Care and Use Committee of the Non-Clinical Research Institute of Corestem ChemOn Co., Ltd. based on the Animal Protection Act and the Laboratory Animals Act (Review No.: 2024-0110).

[0067] The pain level was maximally reduced within 30 minutes of administration of the perilla leaf extract, after which the pain level tended to worsen. This confirmed that the perilla leaf extract had the greatest effect within 30 minutes. In addition, it was confirmed that the pain level was reduced compared to the vehicle control group when administered with the perilla leaf extract, and when comparing the results when administered at 30 and 50 mg / kg.bw., it was confirmed that the anti-pain effect was dose-dependent (Fig. 2A).

[0068] After two weeks of oral administration of perilla leaf extract, the degree of swelling in the hind paws was severe in the control group, but a tendency to recover was observed in the group administered 50 mg / kg.bw of perilla leaf extract (Fig. 2B).

Claims

1. A pharmaceutical composition for pain relief containing perilla leaf extract as an active ingredient.

2. In paragraph 1, A pharmaceutical composition characterized in that the above perilla leaf extract is extracted with 70% ethanol.

3. In paragraph 1, A pharmaceutical composition characterized in that the above perilla leaf extract is extracted including a blanching process prior to extraction.

4. In paragraph 1, A pharmaceutical composition characterized in that the above perilla leaf extract exhibits an effect through a mechanism of inhibiting NO production or suppressing cytokine expression.

5. In paragraph 4, A pharmaceutical composition characterized in that the cytokine is at least one selected from the group consisting of IL-1β, IL-6, GM-CSF, and TNF-α.

6. A method for inhibiting the expression of at least one cytokine selected from the group consisting of IL-1β, IL-6, GM-CSF, and TNF-α, comprising administering to a subject a pharmaceutical composition according to any one of claims 1 to 5.

7. In paragraph 6, A method characterized in that the pharmaceutical composition suppresses cytokines IL-1β, IL-6, GM-CSF and TNF-α.

8. Health functional food composition for pain relief containing perilla leaf extract.

9. Feed composition for pain relief containing perilla leaf extract.

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