WO2018080054A1 - Composition for preventing, alleviating or treating arthritis, containing extract of mollugo pentaphylla as active ingredient - Google Patents

Composition for preventing, alleviating or treating arthritis, containing extract of mollugo pentaphylla as active ingredient Download PDF

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WO2018080054A1
WO2018080054A1 PCT/KR2017/011224 KR2017011224W WO2018080054A1 WO 2018080054 A1 WO2018080054 A1 WO 2018080054A1 KR 2017011224 W KR2017011224 W KR 2017011224W WO 2018080054 A1 WO2018080054 A1 WO 2018080054A1
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arthritis
pomegranate
extract
composition
preventing
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PCT/KR2017/011224
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French (fr)
Korean (ko)
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김동선
이윤미
손은정
김온순
이영실
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한국 한의학 연구원
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A23L33/105—Plant extracts, their artificial duplicates or their derivatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/36—Caryophyllaceae (Pink family), e.g. babysbreath or soapwort
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2200/00—Function of food ingredients
    • A23V2200/30—Foods, ingredients or supplements having a functional effect on health
    • A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention

Definitions

  • the present invention is a pomegranate grass ( Mollugo pentaphylla ) relates to a composition for the prevention, improvement or treatment of arthritis containing the extract as an active ingredient.
  • inflammatory diseases including arthritis
  • Inflammatory diseases caused by these inflammatory reactions include gastritis, colitis, arthritis, nephritis, hepatitis, arteriosclerosis, cancer or degenerative diseases.
  • effective drugs or treatments for the prevention and treatment of arthritis diseases have not been developed.
  • Arthritis refers to the development of inflammatory changes in the joints due to various causes such as aging, mechanical damage, and immune disorders.
  • osteoarthritis is a type of arthritis, also called degenerative arthritis, and refers to arthritis caused by degenerative changes in cartilage and surrounding bone in lubricated joints.
  • osteoarthritis is a disease characterized by gradual loss of articular cartilage, hypertrophy of bone located below the cartilage, bone formation at the edge of the joint, and nonspecific synovial inflammation.
  • Osteoarthritis is a disease caused by cartilage damage caused by aging or excessive physical pressure (eg, obesity, trauma, etc.). Therefore, osteoarthritis represents severe pain and movement disorders such as knee joints, knee joints, hip joints, etc., which are heavily weighted, resulting in deformation of joints when left for a long time.
  • gouty arthritis is an inflammation caused by the deposition of uric acid in the spaces and tissues of the joints. Urate is increased by increasing the concentration of uric acid (the product of the body's metabolism of purine, which is obtained through food). (Uric acid is in the form of urate in the blood, body fluids, and joint fluids.) Crystals deposit in the cartilage, tendons, and surrounding tissue of the joint. This phenomenon causes inflammation of the joints, leading to recurrent attacks with extreme pain.
  • pomegranate grass grows in a field or vacant land or roadside, the stem is thin, many branches are split in the upper part, the height is 10-30cm, hairless, there is a row of hairs.
  • the leaves at the bottom of the stem have 3 to 5 turns, and the leaves at the upper part face each other and have a basso shape or upside down basso shape with flat edges, narrow ends, and no petioles.
  • Flower is yellowish green in July ⁇ October and runs in the form of pickling inflorescence at the end of branch and leaf axil.
  • a bract is membranous ( ⁇ : translucent like a thin sheet of paper), and a small peduncle sags downward after flowering.
  • Petals are absent, 5 calyx pieces, 1.8mm long ellipse shape. Surgery is 3 ⁇ 5, pistil is 1, pistil is 3, short. Fruits are capsules, round, divided into three. Seeds are flat kidneys, dark brown, with papillae. Pomegranate leaves are similar to pomegranate trees. It is widely distributed in temperate and tropical regions of Asia, including Korea.
  • Korean Patent Publication No. 2016-0011382 discloses a pharmaceutical composition for preventing or treating cancer and a health functional food containing the large pomegranate grass extract as an active ingredient.
  • No. 2016-0001805 discloses a composition for the prevention, amelioration or treatment of Tbili2-mediated autoimmune diseases comprising pomegranate extract, plantain tack extract, kidney bean extract or horseback riding extract, but the pomegranate extract of the present invention is effective.
  • a composition for preventing, ameliorating or treating arthritis which is contained as a component.
  • the present invention has been made by the above-mentioned demands, and provides a composition for the prevention, improvement or treatment of arthritis containing pomegranate extract as an active ingredient, the pomegranate extract of the present invention is not cytotoxic, due to inflammation It has the effect of inhibiting the production of induced NO, increase of TNF ⁇ , PGE 2 and IL-6, not only reduces the content of IL-1 ⁇ increased by gouty arthritis, but also relieves pain caused by osteoarthritis, and inflammatory cytokines By confirming the effect of reducing the content of, the present invention was completed.
  • the present invention provides a dietary supplement for the prevention or improvement of arthritis containing pomegranate grass extract as an active ingredient.
  • the present invention also provides a pharmaceutical composition for the prevention or treatment of arthritis containing pomegranate extract as an active ingredient.
  • the present invention relates to a composition for the prevention, improvement or treatment of arthritis containing pomegranate extract as an active ingredient, in detail, pomegranate extract of the present invention is not cytotoxic, the production of NO induced by inflammation, TNF ⁇ It has the effect of inhibiting the increase of PGE 2 and IL-6, and reduces the amount of IL-1 ⁇ increased by gouty arthritis, as well as alleviating the pain caused by osteoarthritis, and reducing the content of inflammatory cytokines. There is.
  • Figure 2 shows that the production of NO increased by the treatment of LPS (lipopolysaccharide) in RAW 264.7 cells, the result of confirming that the production of increased NO by LPS by treating the pomegranate extract of the present invention.
  • LPS lipopolysaccharide
  • Figure 2 shows that the production of NO increased by the treatment of LPS (lipopolysaccharide) in RAW 264.7 cells, the result of confirming that the production of increased NO by LPS by treating the pomegranate extract of the present invention.
  • #### means that NO production increased significantly compared to the negative control by LPS treatment, which means that p ⁇ 0.0001, and * and *** indicate that NO production increased by LPS.
  • Treatment of pomegranate extract was statistically significant, with * being p ⁇ 0.05 and *** being p ⁇ 0.001.
  • MSU monosodium ureate
  • Con is a control group that did not induce gouty arthritis
  • MSU is a group caused by gouty arthritis by administration of MSU
  • MeOH is MSU + pomegranate pool methanol extract treatment group
  • EtOH is MSU + pomegranate pool ethanol extract treatment group
  • Water is the MSU + pomegranate water extract treatment group
  • colchicine is the positive control group.
  • # Indicates that IL-1 ⁇ content was significantly increased in the group induced by gouty arthritis by MSU administration compared to the normal group, and p ⁇ 0.00001.
  • *, **, *** is that the pomegranate extract treatment group of the present invention compared to the induction group of gouty arthritis statistically significantly reduced IL-1 ⁇ content, * is p ⁇ 0.05, ** Is p ⁇ 0.001 and *** means p ⁇ 0.0001.
  • Figure 7 is the result of confirming the weight load rate (%) in SD rats induced osteoarthritis by MIA (monosodium iodoacetate).
  • Control is a normal group that did not induce osteoarthritis as a control group
  • MIA is a group in which osteoarthritis was induced by MIA administration
  • MIA + Indomethacin (1mg / kg) is a positive control group
  • MIA + pomegranate pool (300mg / kg) Is a group treated with 300 mg / kg pomegranate extract in the osteoarthritis-induced group
  • MIA + pomegranate pool (150 mg / kg) is a group treated with 150 mg / kg pomegranate extract in the osteoarthritis-induced group
  • 75 mg / kg is a group treated with 75 mg / kg pomegranate extract in the osteoarthritis-induced group.
  • **, *** indicates that the pomegranate grass extract treatment group of the present invention compared to
  • FIG. 8 shows the results of confirming changes in expression levels of TNF- ⁇ (A) and IL-6 (B) proteins in SD rats induced with osteoarthritis by MIA (monosodium iodoacetate).
  • Control is a normal group that did not cause osteoarthritis as a control group
  • MIA is a group in which osteoarthritis was induced by MIA administration
  • MIA + Indomethacin (1mg / kg) is a positive control
  • MIA + pomegranate pool (mg / kg) Is a group treated with pomegranate grass extract by concentration in osteoarthritis-induced group.
  • the present invention is a pomegranate grass ( Mollugo pentaphylla ) relates to a health functional food composition for the prevention or improvement of arthritis containing the extract as an active ingredient.
  • the arthritis is osteoarthritis or gouty arthritis, but is not limited thereto.
  • the composition may be characterized by inhibiting inflammatory cytokines and pain, and the pain may be pain due to arthritis.
  • the pomegranate extract may be prepared by a method comprising the following steps, but is not limited thereto:
  • the extraction solvent in step 1) is preferably water, lower alcohols of C 1 ⁇ C 4 or mixtures thereof, more preferably methanol or ethanol extract, but is not limited thereto.
  • the extraction of pomegranate grass may use all conventional methods known in the art, such as filtration, hot water extraction, dipping extraction, reflux cooling extraction, and ultrasonic extraction.
  • Concentration under reduced pressure of step 3) is preferably a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto.
  • the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.
  • composition is preferably, but not limited to, prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrups and beverages.
  • the pomegranate grass extract may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method.
  • the blending amount of the active ingredient can be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment).
  • the composition of the present invention is added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight based on the raw materials.
  • the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety.
  • the kind of food There is no particular limitation on the kind of food.
  • Examples of foods to which the extract or fractions thereof may be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea , Drinks, alcoholic beverages and vitamin complexes, and includes all the health functional foods in the conventional sense.
  • composition of the present invention When the composition of the present invention is used as a health beverage, various flavors, natural carbohydrates, and the like may be contained as additional components, as in general beverages.
  • natural carbohydrates are sugars such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as textine and cyclotenstrin, xylitol, sorbitol and erythritol.
  • sweetening agent natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used.
  • the ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention.
  • the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages.
  • the composition of the present invention may contain a pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical, but the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.
  • the present invention also relates to a pharmaceutical composition for the prevention or treatment of arthritis containing pomegranate extract as an active ingredient.
  • the arthritis is preferably, but not limited to, osteoarthritis or gouty arthritis.
  • composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient, and may be various oral or parenteral formulations.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid preparations for oral administration include capsules, powders, granules, tablets, pills, and the like, which may comprise at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin.
  • lubricants such as magnesium stearate, talc and the like are also used.
  • Liquid preparations for oral administration include suspensions, emulsions, syrups, aerosols and the like, and may include various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to commonly used simple diluents such as water and liquid paraffin.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • the non-aqueous solvent and the suspension solvent propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used.
  • the base of the suppository As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycero gelatin and the like can be used.
  • parenteral administration it is desirable to select either external skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections, most preferably for external skin use.
  • composition according to the invention is administered in a pharmaceutically effective amount.
  • pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, the effective amount of the level of the disease, the severity, the drug activity of the patient , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of the drug, and other factors well known in the medical arts.
  • the compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with a minimum amount without side effects, which can be readily determined by one skilled in the art.
  • the dosage of the composition of the present invention varies depending on the weight, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage is the amount of pomegranate extract It is 0.01-2,000 mg / kg, preferably 30-500 mg / kg, more preferably 50-300 mg / kg, and may be administered 1 to 6 times per day.
  • the compositions of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.
  • RAW 264.7 cells which are mouse macrophage lines, were dispensed in 96-well plates, and the pomegranate ethanol extract prepared in Example 1 was treated at different concentrations (10, 50, 100, 200 ⁇ g / ml) for 24 hours. Or incubated for 48 hours. After culturing the cells, the MTT solution was added and reacted for 4 hours, and then the absorbance of 540 nm was measured using an ELISA reader. The relative cell viability was calculated by comparison with the control group.
  • the pomegranate grass ethanol extract was confirmed that the cytotoxicity does not affect the cell viability.
  • LPS 400 ng / ml
  • RAW 274.7 cells divided into 48-well plates at an appropriate concentration
  • the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (25, 50, 100, 200 ⁇ g / ml). Treated and incubated for 24 hours. After taking the culture supernatant, the amount of NO was measured according to the manufacturer's method using Griess reagent.
  • the amount of NO produced increased by LPS was reduced in a concentration-dependent manner by treatment of pomegranate ethanol extract.
  • LPS 400 ng / ml
  • the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (10, 50, 100, 200 ⁇ g / ml). Treated and incubated for 24 hours. After taking the culture supernatant TNF ⁇ production was measured using the TNF ⁇ ELISA kit according to the manufacturer's method.
  • the amount of TNF ⁇ production increased by LPS was reduced by 96.9% or more by treatment of pomegranate ethanol extract.
  • LPS 400 ng / ml
  • the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (25, 50, 100, 200 ⁇ g / ml). Treated and incubated for 24 hours. After the culture supernatant was taken, the amount of PGE 2 was measured using the PGE 2 ELISA kit according to the manufacturer's method.
  • the amount of PGE 2 produced by LPS was decreased by about 40% when the 40 ⁇ g / ml pomegranate ethanol extract was treated, and the 200 ⁇ g / ml pomegranate ethanol extract was reduced. In the case of treatment, PGE 2 production decreased by more than 99%.
  • LPS 400 ng / ml
  • the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (25, 50, 100, 200 ⁇ g / ml). Treated and incubated for 24 hours. After taking the culture supernatant, the amount of IL-6 produced was measured according to the manufacturer's method using the IL-6 ELISA kit.
  • the amount of IL-6 produced by LPS was decreased in a concentration-dependent manner by treatment of pomegranate ethanol extract.
  • Example 3 The efficacy of seats ryupul extracts from gouty arthritis animal model induced by MSU (Monosodium ureate)
  • Example 1 In order to confirm the efficacy of the pomegranate extract extracted in Example 1 in an animal model of gouty arthritis disease induced by MSU (Monosodium ureate), the effect of reducing the inflammatory cytokine IL-1 ⁇ was confirmed.
  • the pomegranate grass methanol extract, pomegranate grass 70% (v / v) ethanol extract and pomegranate grass water extract were administered orally to 7-week-old C57BL6 mice at concentrations of 150 and 300 mg / kg, respectively.
  • One hour after administration of the pomegranate extract 4 mg of MSU was suspended in 50 ⁇ l of PBS containing 2.5% Tween 80 and injected into the right paw tissue to induce gouty arthritis.
  • the drug was administered once a day for 4 days, followed by necropsy on the 5th day of MSU induction.
  • the paw tissue obtained from the autopsy was immersed in RIPA buffer solution, and the tissue was crushed to measure the expression level of IL-1 ⁇ protein, which is an inflammatory marker in the supernatant, by ELISA, and cholchicine as a positive control.
  • IL-1 ⁇ protein which is an inflammatory marker in the supernatant
  • ELISA e.g., IL-1 ⁇ protein
  • cholchicine e.g., cholchicine
  • IL-1 ⁇ increased due to gouty arthritis was significantly reduced by administration of pomegranate extract.
  • Example 4 the pomegranate grass 70% (v / v) ethanol extract prepared in Example 1 was confirmed the efficacy in the osteoarthritis disease animal model induced by MIA (monosodium iodoacetate). Seven-week-old SD rats were orally administered with pomegranate ethanol extract at 75, 150, and 300 mg / kg concentrations for 3 days. Anesthesia was administered on day 4 (0 day) after administration of the drug, followed by clean hair removal around the right knee joint of the SD rat, and 50 ⁇ l (60 mg / ml) of MIA, a osteoarthritis-inducing substance, diluted with 0.9% saline in the joint cavity. It was.
  • MIA monosodium iodoacetate
  • the pomegranate ethanol extract was orally administered once a day for 14 days, and then autopsied on the 14th day of administration.
  • Indometacin was used as a positive control, and pomegranate extract was administered in the morning and measured in the afternoon, and hind limb weight measurement was performed on -3, 0, 8 and 11 days after MIA. .
  • Hind limb weight load was measured using a foot weigher (Incapacitance tester, Linton instrument Co., UK, Ser No. 01/45/25). In osteoarthritis-induced rats in the tester's holder, the pain of the osteoarthritis stood on the normal foot without MIA, so the weight of both feet was unbalanced and the weight of the MIA-treated foot was relatively light. When measuring the weight of the feet, the weight (g) of both feet was measured without the SD rat's belly touching the sensor of the device.
  • the weight load rate (%) was calculated.
  • the weight load is a force supported by the foot, the weight of both feet in the normal case, the weight load rate (%) appears as 100%, but the pain is worsened, the weight load rate (%) is lowered.
  • % Weight load (weight of hind limb with arthritis / weight of normal hind limb) ⁇ 100
  • the weight-bearing rate was significantly reduced by MIA induction, but when the pomegranate extract of the present invention was administered by concentration after MIA induction, the weight-bearing rate (%) was increased compared to the MIA-induced group. I could confirm it.

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Abstract

The present invention relates to a composition for preventing, alleviating or treating arthritis, containing an extract of Mollugo pentaphylla as an active ingredient. More specifically, the Mollugo pentaphylla extract of the present invention is not cytotoxic, has an effect of inhibiting the generation of NO, induced by inflammation, and the increase in TNFα, PGE2, and IL-6, reduces the IL-1β amount increased because of gouty arthritis, relieves pain caused by osteoarthritis, and reduces the amount of inflammatory cytokines, thereby being usable in the prevention, alleviation or treatment of arthritis.

Description

석류풀 추출물을 유효성분으로 함유하는 관절염의 예방, 개선 또는 치료용 조성물Composition for the prevention, improvement or treatment of arthritis containing pomegranate extract as an active ingredient
본 발명은 석류풀(Mollugo pentaphylla) 추출물을 유효성분으로 함유하는 관절염의 예방, 개선 또는 치료용 조성물에 관한 것이다.The present invention is a pomegranate grass ( Mollugo pentaphylla ) relates to a composition for the prevention, improvement or treatment of arthritis containing the extract as an active ingredient.
노령화 사회로 접어들면서 관절염을 포함하는 염증성 질환이 성별에 상관없이 사회적으로 대두되고 있다. 이러한 염증반응에 의해 발생하는 염증성 질환에는 위염, 대장염, 관절염, 신장염, 간염, 동맥경화, 암 또는 퇴행성 질환 등이 포함된다. 그 중 현재까지 관절염 질환의 예방 및 치료를 위한 효과적인 약제나 치료법은 개발되지 못하고 있다. 관절염은 노화, 기계적 손상, 면역이상 등 다양한 원인에 의해 관절 내에 염증성 변화가 생긴 것을 지칭한다.As we move into an aging society, inflammatory diseases, including arthritis, are emerging socially regardless of gender. Inflammatory diseases caused by these inflammatory reactions include gastritis, colitis, arthritis, nephritis, hepatitis, arteriosclerosis, cancer or degenerative diseases. To date, effective drugs or treatments for the prevention and treatment of arthritis diseases have not been developed. Arthritis refers to the development of inflammatory changes in the joints due to various causes such as aging, mechanical damage, and immune disorders.
상기한 관절염 중에서, 골관절염(osteoarthritis)은 퇴행성 관절염으로 칭해지기도 하는 관절염의 일종으로서, 윤활 관절에서 연골과 주위골에 퇴행성 변화가 나타나서 생기는 관절염을 말한다. 즉, 골관절염은 관절 연골의 점차적인 소실과 더불어 연골 하방에 위치한 뼈의 비대, 관절 가장자리 부위의 골 생성, 및 비특이적인 활막 염증을 특징으로 하는 질환이다. 골관절염은 노화나 과도한 물리적 압박(예를 들어, 비만, 외상 등)에 의해서 연골이 손상되어 발생하는 질환이다. 따라서, 골관절염은 체중을 많이 받는 관절, 즉, 무릎(슬)관절, 엉덩이 (고)관절 등에 심한 통증과 운동 장애를 나타내며, 장기간 방치할 경우에는 관절의 변형까지 초래하게 된다.Of the above arthritis, osteoarthritis is a type of arthritis, also called degenerative arthritis, and refers to arthritis caused by degenerative changes in cartilage and surrounding bone in lubricated joints. In other words, osteoarthritis is a disease characterized by gradual loss of articular cartilage, hypertrophy of bone located below the cartilage, bone formation at the edge of the joint, and nonspecific synovial inflammation. Osteoarthritis is a disease caused by cartilage damage caused by aging or excessive physical pressure (eg, obesity, trauma, etc.). Therefore, osteoarthritis represents severe pain and movement disorders such as knee joints, knee joints, hip joints, etc., which are heavily weighted, resulting in deformation of joints when left for a long time.
또한, 통풍성 관절염은 관절 내 공간과 조직에 요산이 침착되면서 발생하는 염증으로, 혈액 내에 요산(음식을 통해 섭취되는 퓨린(purine)이라는 물질을 인체가 대사하고 남은 산물)의 농도가 높아지면서 요산염(요산이 혈액, 체액, 관절액 내에서는 요산염의 형태 존재함) 결정이 관절의 연골, 힘줄, 주위 조직에 침착되는 질병이다. 이러한 현상은 관절의 염증을 유발하여 극심한 통증을 동반하는 재발성 발작을 일으킨다.In addition, gouty arthritis is an inflammation caused by the deposition of uric acid in the spaces and tissues of the joints. Urate is increased by increasing the concentration of uric acid (the product of the body's metabolism of purine, which is obtained through food). (Uric acid is in the form of urate in the blood, body fluids, and joint fluids.) Crystals deposit in the cartilage, tendons, and surrounding tissue of the joint. This phenomenon causes inflammation of the joints, leading to recurrent attacks with extreme pain.
한편, 석류풀은 밭이나 빈터, 또는 길가에서 자라는데, 줄기는 가늘고 윗부분에서 많은 가지가 갈라지며 높이가 10∼30cm이고 털이 없으며 모가 난 줄이 있다. 줄기 밑 부분의 잎은 3∼5개가 돌려나고, 윗부분의 잎은 마주나며 바소 모양 또는 거꾸로 세운 바소 모양이고 가장자리가 밋밋하며 양끝이 좁고 잎자루가 없다. 꽃은 7∼10월에 노란 색을 띤 녹색으로 피고 가지 끝과 잎겨드랑이에 취산꽃차례를 이루며 달린다. 포는 막질(膜質: 얇은 종이처럼 반투명한 것)이고, 작은 꽃자루는 꽃이 진 다음 밑으로 처진다. 꽃잎은 없고 꽃받침조각은 5개이며 길이 1.8mm의 긴 타원 모양이다. 수술은 3∼5개이고, 암술은 1개이며 암술대는 3개이고 짧다. 열매는 삭과이고 둥글며 3개로 갈라진다. 종자는 편평한 콩팥 모양이고 진한 갈색이며 잔돌기가 있다. 잎이 석류나무와 비슷하여 석류풀이라고 한다. 한국을 비롯하여 아시아의 온대와 열대에 널리 분포한다. On the other hand, pomegranate grass grows in a field or vacant land or roadside, the stem is thin, many branches are split in the upper part, the height is 10-30cm, hairless, there is a row of hairs. The leaves at the bottom of the stem have 3 to 5 turns, and the leaves at the upper part face each other and have a basso shape or upside down basso shape with flat edges, narrow ends, and no petioles. Flower is yellowish green in July ~ October and runs in the form of pickling inflorescence at the end of branch and leaf axil. A bract is membranous (膜質: translucent like a thin sheet of paper), and a small peduncle sags downward after flowering. Petals are absent, 5 calyx pieces, 1.8mm long ellipse shape. Surgery is 3 ~ 5, pistil is 1, pistil is 3, short. Fruits are capsules, round, divided into three. Seeds are flat kidneys, dark brown, with papillae. Pomegranate leaves are similar to pomegranate trees. It is widely distributed in temperate and tropical regions of Asia, including Korea.
석류풀 추출물 관련 기술의 일례로는 한국공개특허 제2016-0011382호에 큰석류풀 추출물을 유효성분으로 함유하는 암 예방 또는 치료용 약학적 조성물 및 건강기능식품에 관한 것이 개시되어 있고, 한국공개특허 제2016-0001805호에 석류풀 추출물, 질경이택사 추출물, 담팥수 추출물 또는 개승마 추출물을 포함하는 Ｔｈ2-매개 면역 질환의 예방, 개선 또는 치료용 조성물이 개시되어 있으나, 본 발명의 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방, 개선 또는 치료용 조성물에 대해 개시된 바 없다.As an example of pomegranate grass extract related technology, Korean Patent Publication No. 2016-0011382 discloses a pharmaceutical composition for preventing or treating cancer and a health functional food containing the large pomegranate grass extract as an active ingredient. No. 2016-0001805 discloses a composition for the prevention, amelioration or treatment of Tbili2-mediated autoimmune diseases comprising pomegranate extract, plantain tack extract, kidney bean extract or horseback riding extract, but the pomegranate extract of the present invention is effective. There is no disclosure of a composition for preventing, ameliorating or treating arthritis, which is contained as a component.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방, 개선 또는 치료용 조성물을 제공하고, 본 발명의 석류풀 추출물이 세포 독성이 없으며, 염증에 의해 유도되는 NO의 생성, TNFα, PGE2 및 IL-6의 증가를 억제하는 효과가 있고, 통풍성 관절염에 의해 증가한 IL-1β의 함량을 감소시킬 뿐만 아니라, 골관절염에 의한 통증을 완화하며, 염증성 사이토카인의 함량을 감소시키는 효과를 확인함으로써, 본 발명을 완성하였다.The present invention has been made by the above-mentioned demands, and provides a composition for the prevention, improvement or treatment of arthritis containing pomegranate extract as an active ingredient, the pomegranate extract of the present invention is not cytotoxic, due to inflammation It has the effect of inhibiting the production of induced NO, increase of TNFα, PGE 2 and IL-6, not only reduces the content of IL-1β increased by gouty arthritis, but also relieves pain caused by osteoarthritis, and inflammatory cytokines By confirming the effect of reducing the content of, the present invention was completed.
상기 목적을 달성하기 위하여, 본 발명은 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방 또는 개선용 건강기능식품 조성물을 제공한다.In order to achieve the above object, the present invention provides a dietary supplement for the prevention or improvement of arthritis containing pomegranate grass extract as an active ingredient.
또한, 본 발명은 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for the prevention or treatment of arthritis containing pomegranate extract as an active ingredient.
본 발명은 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방, 개선 또는 치료용 조성물에 관한 것으로, 상세하게는 본 발명의 석류풀 추출물은 세포 독성이 없으며, 염증에 의해 유도되는 NO의 생성, TNFα, PGE2 및 IL-6의 증가를 억제하는 효과가 있고, 통풍성 관절염에 의해 증가된 IL-1β의 함량을 감소시킬 뿐만 아니라, 골관절염에 의한 통증을 완화시키며, 염증성 사이토카인의 함량을 감소시키는 효과가 있다.The present invention relates to a composition for the prevention, improvement or treatment of arthritis containing pomegranate extract as an active ingredient, in detail, pomegranate extract of the present invention is not cytotoxic, the production of NO induced by inflammation, TNFα It has the effect of inhibiting the increase of PGE 2 and IL-6, and reduces the amount of IL-1β increased by gouty arthritis, as well as alleviating the pain caused by osteoarthritis, and reducing the content of inflammatory cytokines. There is.
도 1은 본 발명의 석류풀 추출물이 RAW 264.7 세포의 생존률에 미치는 영향을 확인한 결과이다.1 is a result confirming the effect of the pomegranate grass extract of the present invention on the survival rate of RAW 264.7 cells.
도 2는 RAW 264.7 세포에서 LPS(lipopolysaccharide)의 처리에 의해 NO 생성량이 증가하며, 본 발명의 석류풀 추출물을 처리하여 LPS에 의해 증가된 NO의 생성량이 감소하는 것을 확인한 결과이다. ####는 LPS의 처리에 의해 NO 생성이 음성대조구(Control)에 비해 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임의 의미하고, *, ***는 LPS에 의해 증가된 NO 생성량이 석류풀 추출물의 처리하여 통계적으로 유의미하게 감소하였다는 것으로, *는 p<0.05이고, ***는 p<0.001이다.Figure 2 shows that the production of NO increased by the treatment of LPS (lipopolysaccharide) in RAW 264.7 cells, the result of confirming that the production of increased NO by LPS by treating the pomegranate extract of the present invention. #### means that NO production increased significantly compared to the negative control by LPS treatment, which means that p <0.0001, and * and *** indicate that NO production increased by LPS. Treatment of pomegranate extract was statistically significant, with * being p <0.05 and *** being p <0.001.
도 3은 RAW 264.7 세포에서 LPS(lipopolysaccharide)에 의해 유도된 TNFα 생성량을 본 발명의 석류풀 추출물을 처리하여 TNFα의 생성량을 감소시키는 것을 확인한 결과이다.3 is a result of confirming that the TNFα production induced by LPS (lipopolysaccharide) in RAW 264.7 cells reduces the production of TNFα by treating the pomegranate extract of the present invention.
도 4는 RAW 264.7 세포에서 LPS(lipopolysaccharide)에 의해 유도된 PGE2 생성량을 본 발명의 석류풀 추출물을 처리하여 PGE2의 생성량을 감소시키는 것을 확인한 결과이다. ####는 LPS의 처리에 의해 PGE2 생성이 음성대조구(Control)에 비해 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임의 의미하고, *, ***는 LPS에 의해 증가된 PGE2 생성량이 석류풀 추출물의 처리하여 통계적으로 유의미하게 감소하였다는 것으로, *는 p<0.05이고, ***는 p<0.001이다.4 is a result of confirming that the production of PGE 2 induced by lipopolysaccharide (LPS) in RAW 264.7 cells reduces the production of PGE 2 by treating the pomegranate extract of the present invention. #### means that PGE 2 production was increased statistically significantly compared to the negative control (Control) by the treatment of LPS, p <0.0001, *, *** is PGE 2 increased by LPS Production was statistically significantly reduced by treatment of pomegranate extract, where * is p <0.05 and *** is p <0.001.
도 5는 RAW 264.7 세포에서 LPS에 의해 유도된 IL-6 생성량을 본 발명의 석류풀 추출물을 처리하여 IL-6의 생성량을 감소시키는 것을 확인한 결과이다. ####는 LPS의 처리에 의해 IL-6 생성이 음성대조구(Control)에 비해 통계적으로 유의미하게 증가하였다는 것으로, p<0.0001임의 의미하고, **, ***는 LPS에 의해 증가된 IL-6 생성량이 석류풀 추출물의 처리하여 통계적으로 유의미하게 감소하였다는 것으로, **는 p<0.01이고, ***는 p<0.001이다.5 is a result of confirming that the production of IL-6 induced by LPS in RAW 264.7 cells reduces the production of IL-6 by treating the pomegranate extract of the present invention. #### means that IL-6 production was statistically significantly increased compared to the negative control (Control) by the treatment of LPS, meaning that p <0.0001, **, *** is increased by LPS IL-6 production was statistically significantly reduced by treatment of pomegranate extract, ** is p <0.01, *** is p <0.001.
도 6은 MSU(monosodium ureate)에 의해 통풍성 관절염이 유도된 C57BL6 마우스에서 IL-1β의 감소효과를 확인한 결과이다. Con은 대조군으로 통풍성 관절염을 유발시키지 않은 정상군이며, MSU는 MSU의 투여에 의해 통풍성 관절염이 유발한 군이고, MeOH는 MSU+석류풀 메탄올 추출물 처리군이고, EtOH는 MSU+석류풀 에탄올 추출물 처리군이며, Water는 MSU+석류풀 물 추출물 처리군이고, colchicine은 양성대조군이다. #는 MSU 투여에 의해 통풍성 관절염이 유발한 군이 정상군에 비해 통계적으로 유의미하게 IL-1β의 함량이 증가하였다는 것으로, p<0.00001임을 의미한다. *, **, ***는 통풍성 관절염의 유발군에 대비하여 본 발명의 석류풀 추출물 처리군이 통계적으로 유의미하게 IL-1β의 함량이 감소하였다는 것으로, *는 p<0.05이고, **는 p<0.001이며, ***는 p<0.0001임을 의미한다.6 is a result confirming the reduction effect of IL-1β in C57BL6 mice induced gouty arthritis by MSU (monosodium ureate). Con is a control group that did not induce gouty arthritis, MSU is a group caused by gouty arthritis by administration of MSU, MeOH is MSU + pomegranate pool methanol extract treatment group, EtOH is MSU + pomegranate pool ethanol extract treatment group , Water is the MSU + pomegranate water extract treatment group and colchicine is the positive control group. # Indicates that IL-1β content was significantly increased in the group induced by gouty arthritis by MSU administration compared to the normal group, and p <0.00001. *, **, *** is that the pomegranate extract treatment group of the present invention compared to the induction group of gouty arthritis statistically significantly reduced IL-1β content, * is p <0.05, ** Is p <0.001 and *** means p <0.0001.
도 7은 MIA(monosodium iodoacetate)에 의해 골 관절염이 유도된 SD 랫트에서 체중부하율(%)를 확인한 결과이다. Control은 대조군으로 골 관절염을 유발시키지 않은 정상군이며, MIA는 MIA의 투여에 의해 골 관절염이 유발한 군이고, MIA+Indomethacin(1mg/kg)은 양성대조군이고, MIA+석류풀(300mg/kg)은 골 관절염 유발군에 300mg/kg의 석류풀 추출물을 처리한 군이고, MIA+석류풀(150mg/kg)은 골 관절염 유발군에 150mg/kg의 석류풀 추출물을 처리한 군이며, MIA+석류풀(75mg/kg)은 골 관절염 유발군에 75mg/kg의 석류풀 추출물을 처리한 군이다. **, ***는 골 관절염 유발군에 대비하여 본 발명의 석류풀 추출물 처리군이 통계적으로 유의미하게 체중부하율(%)이 증가하였다는 것으로, **는 p<0.01, ***는 p<0.001임을 의미한다.Figure 7 is the result of confirming the weight load rate (%) in SD rats induced osteoarthritis by MIA (monosodium iodoacetate). Control is a normal group that did not induce osteoarthritis as a control group, MIA is a group in which osteoarthritis was induced by MIA administration, MIA + Indomethacin (1mg / kg) is a positive control group, and MIA + pomegranate pool (300mg / kg) Is a group treated with 300 mg / kg pomegranate extract in the osteoarthritis-induced group, MIA + pomegranate pool (150 mg / kg) is a group treated with 150 mg / kg pomegranate extract in the osteoarthritis-induced group, 75 mg / kg) is a group treated with 75 mg / kg pomegranate extract in the osteoarthritis-induced group. **, *** indicates that the pomegranate grass extract treatment group of the present invention compared to the osteoarthritis-induced group significantly increased the weight load (%), ** is p <0.01, *** is p Means <0.001.
도 8은 MIA(monosodium iodoacetate)에 의해 골 관절염이 유도된 SD 랫트에서 TNF-α(A) 및 IL-6 (B) 단백질의 발현량의 변화를 확인한 결과이다. Control은 대조군으로 골 관절염을 유발시키지 않은 정상군이며, MIA는 MIA의 투여에 의해 골 관절염이 유발한 군이고, MIA+Indomethacin(1mg/kg)은 양성 대조군이며, MIA+석류풀(mg/kg)은 골 관절염 유발군에 농도별로 석류풀 추출물을 처리한 군이다. #, ##는 MIA 투여군이 정상군(Control)에 비해 통계적으로 유의미하게 TNF-α 및 IL-6 단백질의 발현량이 증가하였다는 것으로, #는 p<0.05이고, ##는 p<0.01임을 의미한다. *, ***는 MIA 투여에 의해 골 관절염이 유발된 군에 대비하여 본 발명의 석류풀 추출물 처리군이 통계적으로 유의미하게 TNF-α 및 IL-6 단백질의 발현량이 감소하였다는 것으로, *는 p<0.05이고, ***는 p<0.0001임을 의미한다.FIG. 8 shows the results of confirming changes in expression levels of TNF-α (A) and IL-6 (B) proteins in SD rats induced with osteoarthritis by MIA (monosodium iodoacetate). Control is a normal group that did not cause osteoarthritis as a control group, MIA is a group in which osteoarthritis was induced by MIA administration, MIA + Indomethacin (1mg / kg) is a positive control, MIA + pomegranate pool (mg / kg) Is a group treated with pomegranate grass extract by concentration in osteoarthritis-induced group. #, ## indicates that the TIAF-α and IL-6 protein levels were significantly increased in the MIA-treated group compared to the control group, where # is p <0.05 and ## is p <0.01. do. *, *** indicates that the pomegranate extract treatment group of the present invention significantly reduced the expression levels of TNF-α and IL-6 protein compared to the group in which osteoarthritis was induced by MIA administration. p <0.05 and *** means p <0.0001.
본 발명은 석류풀(Mollugo pentaphylla) 추출물을 유효성분으로 함유하는 관절염의 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.The present invention is a pomegranate grass ( Mollugo pentaphylla ) relates to a health functional food composition for the prevention or improvement of arthritis containing the extract as an active ingredient.
상기 관절염은 골 관절염 또는 통풍성 관절염인 것이 바람직하지만 이에 한정하지 않으며, 상기 조성물은 염증성 사이토카인 및 통증을 억제하는 것이 특징이며, 상기 통증은 관절염에 의한 통증일 수 있다.Preferably, the arthritis is osteoarthritis or gouty arthritis, but is not limited thereto. The composition may be characterized by inhibiting inflammatory cytokines and pain, and the pain may be pain due to arthritis.
상기 석류풀 추출물은 하기의 단계를 포함하는 방법에 의해 제조되는 것일 수 있으나, 이에 한정하지 않는다:The pomegranate extract may be prepared by a method comprising the following steps, but is not limited thereto:
1) 석류풀 식물에 추출용매를 가하여 추출하는 단계;1) extracting by adding an extraction solvent to the pomegranate plant;
2) 단계 1)의 추출물을 여과하는 단계; 및2) filtering the extract of step 1); And
3) 단계 2)의 여과한 추출물을 감압 농축하고 건조하여 추출물을 제조하는 단계.3) concentration of the filtered extract of step 2) under reduced pressure and drying to prepare an extract.
상기 단계 1)에서 추출용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물인 것이 바람직하며, 더 바람직하게는 메탄올 또는 에탄올 추출물이지만 이에 한정하지 않는다. The extraction solvent in step 1) is preferably water, lower alcohols of C 1 ~ C 4 or mixtures thereof, more preferably methanol or ethanol extract, but is not limited thereto.
상기 제조방법에 있어서, 석류풀의 추출은 여과법, 열수 추출, 침지 추출, 환류 냉각 추출 및 초음파 추출 등의 당 업계에 공지된 모든 통상적인 방법을 이용할 수 있다. 상기 단계 3)의 감압농축은 진공 감압 농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무 건조 또는 동결 건조하는 것이 바람직하나 이에 한정하지 않는다.In the above production method, the extraction of pomegranate grass may use all conventional methods known in the art, such as filtration, hot water extraction, dipping extraction, reflux cooling extraction, and ultrasonic extraction. Concentration under reduced pressure of step 3) is preferably a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.
상기 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조되는 것이 바람직하지만 이에 제한하는 것은 아니다.The composition is preferably, but not limited to, prepared in any one formulation selected from powders, granules, pills, tablets, capsules, candy, syrups and beverages.
본 발명의 건강기능식품 조성물을 식품첨가물로 사용하는 경우, 상기 석류풀 추출물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 유효 성분의 혼합양은 그의 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. 일반적으로, 식품 또는 음료의 제조시에 본 발명의 조성물은 원료에 대하여 15 중량부 이하, 바람직하게는 10 중량부 이하의 양으로 첨가된다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하는 장기간의 섭취의 경우에는 상기 양은 상기 범위 이하일 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다. 상기 식품의 종류에는 특별한 제한은 없다. 상기 추출물 또는 이의 분획물을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 수프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합체 등이 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다. When the health functional food composition of the present invention is used as a food additive, the pomegranate grass extract may be added as it is or used with other food or food ingredients, and may be appropriately used according to a conventional method. The blending amount of the active ingredient can be suitably determined according to the purpose of its use (prevention, health or therapeutic treatment). Generally, in the preparation of food or beverages, the composition of the present invention is added in an amount of up to 15 parts by weight, preferably up to 10 parts by weight based on the raw materials. However, in the case of long-term intake for health and hygiene or health control purposes, the amount may be below the above range, and the active ingredient may be used in an amount above the above range because there is no problem in terms of safety. There is no particular limitation on the kind of food. Examples of foods to which the extract or fractions thereof may be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, drinks, tea , Drinks, alcoholic beverages and vitamin complexes, and includes all the health functional foods in the conventional sense.
본 발명의 조성물을 건강 음료로 사용할 경우, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 텍스트린, 사이클로텐스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100g당 일반적으로 약 0.01~0.04g, 바람직하게는 약 0.02~0.03g이다. 상기 외에 본 발명의 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 중점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 혼합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 조성물은 100 중량부 당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.When the composition of the present invention is used as a health beverage, various flavors, natural carbohydrates, and the like may be contained as additional components, as in general beverages. The above-mentioned natural carbohydrates are sugars such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as textine and cyclotenstrin, xylitol, sorbitol and erythritol. As the sweetening agent, natural sweetening agents such as tautin and stevia extract, synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 g of the composition of the present invention. In addition to the above, the composition of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusting agents, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages. In addition, the composition of the present invention may contain a pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components can be used independently or in combination. The proportion of such additives is not critical, but the composition of the present invention is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight.
또한, 본 발명은 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방 또는 치료용 약학 조성물에 관한 것이다. 상기 관절염은 골관절염 또는 통풍성 관절염인 것이 바람직하지만 이에 한정하지 않는다.The present invention also relates to a pharmaceutical composition for the prevention or treatment of arthritis containing pomegranate extract as an active ingredient. The arthritis is preferably, but not limited to, osteoarthritis or gouty arthritis.
본 발명의 조성물은 상기 유효성분 이외에 약학적으로 허용 가능한 담체, 부형제 또는 희석제를 더 포함할 수 있으며, 경구 또는 비경구의 여러 가지 제형일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형 제제에는 캡슐제, 산제, 과립제, 정제, 환제 등이 포함되며, 이러한 고형 제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용된다. 경구 투여를 위한 액상 제제로는 현탁액, 에멀전, 시럽, 에어로졸 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성 용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성 용제 및 현탁 용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로 젤라틴 등이 사용될 수 있다. 비경구 투여 시 피부 외용 또는 복강 내, 직장, 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내 주사 방식을 선택하는 것이 바람직하며, 가장 바람직하게는 피부 외용으로 사용한다.The composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient, and may be various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used. Solid preparations for oral administration include capsules, powders, granules, tablets, pills, and the like, which may comprise at least one excipient such as starch, calcium carbonate, sucrose or lactose (at least one compound). lactose) and gelatin. In addition to simple excipients, lubricants such as magnesium stearate, talc and the like are also used. Liquid preparations for oral administration include suspensions, emulsions, syrups, aerosols and the like, and may include various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc., in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycero gelatin and the like can be used. For parenteral administration, it is desirable to select either external skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections, most preferably for external skin use.
본 발명에 따른 약학 조성물은 약제학적으로 유효한 양으로 투여한다. 본 발명에 있어서, "약제학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효량의 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition according to the invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, the effective amount of the level of the disease, the severity, the drug activity of the patient , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of the drug, and other factors well known in the medical arts. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be single or multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with a minimum amount without side effects, which can be readily determined by one skilled in the art.
본 발명의 조성물의 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설률 및 질환의 중증도에 따라 그 범위가 다양하며, 일일 투여량은 석류풀 추출물의 양을 기준으로 0.01~2,000mg/kg이고, 바람직하게는 30~500mg/kg이고, 더욱 바람직하게는 50~300mg/kg이며, 하루 1~6회 투여될 수 있다. 본 발명의 조성물은 단독으로 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 및 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.The dosage of the composition of the present invention varies depending on the weight, age, sex, health status, diet, time of administration, administration method, excretion rate and severity of the disease of the patient, the daily dosage is the amount of pomegranate extract It is 0.01-2,000 mg / kg, preferably 30-500 mg / kg, more preferably 50-300 mg / kg, and may be administered 1 to 6 times per day. The compositions of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다. Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for explaining the present invention in more detail, it is obvious to those skilled in the art that the scope of the present invention is not limited by them.
실시예Example 1.  One. 석류풀Pomegranate grass 추출물의 제조 Preparation of Extract
1) 석류풀 물 추출물의 제조1) Preparation of Pomegranate Water Extract
건조된 1kg의 석류풀 전초에 15ℓ의 물을 첨가하여 80℃에서 3시간 동안 환류추출한 후 여과한 액을 60℃에서 감압 농축하였다. 상기 농축액을 건조하여 석류풀 물 추출물(238g)을 제조하였다.15 liters of water was added to the dried 1 kg pomegranate outpost and refluxed at 80 ° C. for 3 hours, and the filtrate was concentrated under reduced pressure at 60 ° C. The concentrate was dried to prepare pomegranate water extract (238 g).
2) 석류풀 메탄올 추출물의 제조2) Preparation of Pomegranate Methanol Extract
건조된 1kg의 석류풀 전초에 15ℓ의 메탄올을 첨가하여 65℃에서 3시간 동안 환류추출한 후, 여과한 액을 40℃에서 감압 농축하였다. 상기 농축액을 건조하여 석류풀 메탄올 추출물(125g)을 제조하였다.15 L of methanol was added to the dried 1 kg pomegranate outpost, and the mixture was refluxed at 65 ° C. for 3 hours, and the filtrate was concentrated under reduced pressure at 40 ° C. The concentrate was dried to prepare a pomegranate grass methanol extract (125 g).
3) 석류풀 에탄올 추출물의 제조3) Preparation of Pomegranate Pool Ethanol Extract
건조된 1kg의 석류풀 전초에 15ℓ의 70%(v/v) 에탄올을 첨가하여 80℃에서 3시간동안 환류추출한 후 여과한 액을 45℃에서 감압 농축하였다. 상기 농축액을 건조하여 석류풀 에탄올 추출물(139g)을 제조하였다.15 L of 70% (v / v) ethanol was added to the dried 1 kg pomegranate outpost, and the mixture was refluxed at 80 ° C. for 3 hours, and the filtrate was concentrated under reduced pressure at 45 ° C. The concentrate was dried to prepare a pomegranate ethanol extract (139 g).
실시예Example 2. RAW 264.7 세포에서  2. In RAW 264.7 Cells 석류풀Pomegranate grass 추출물의 항염증 효능 평가 Evaluation of Anti-inflammatory Effects of Extracts
1) MTT 어세이1) MTT Assay
생쥐 대식세포주인 RAW 264.7 세포를 96웰 플레이트에 적정 농도로 분주한 후, 상기 실시예 1에서 제조한 석류풀 에탄올 추출물을 농도별(10, 50, 100, 200㎍/㎖)로 처리하여 24시간 혹은 48시간 동안 배양하였다. 배양이 끝난 세포에 MTT 용액을 첨가, 4시간 반응시킨 후 ELISA 리더를 이용하여 540nm의 흡광도를 측정하였고, 이후 대조군과의 비교를 통해 상대적인 세포생존율(% of control)을 계산하였다. RAW 264.7 cells, which are mouse macrophage lines, were dispensed in 96-well plates, and the pomegranate ethanol extract prepared in Example 1 was treated at different concentrations (10, 50, 100, 200 µg / ml) for 24 hours. Or incubated for 48 hours. After culturing the cells, the MTT solution was added and reacted for 4 hours, and then the absorbance of 540 nm was measured using an ELISA reader. The relative cell viability was calculated by comparison with the control group.
그 결과 도 1에 개시한 바와 같이 석류풀 에탄올 추출물이 세포독성이 없어 세포생존율에 영향을 미치지 않는다는 것을 확인하였다.As a result, as shown in FIG. 1, the pomegranate grass ethanol extract was confirmed that the cytotoxicity does not affect the cell viability.
2) NO 측정2) NO measurement
48웰 플레이트에 적정 농도로 분주한 RAW 274.7 세포에 LPS(400ng/㎖)를 처리하고, 상기 실시예 1에서 제조한 석류풀 에탄올 추출물을 농도별(25, 50, 100, 200㎍/㎖)로 처리하여 24시간 동안 배양하였다. 배양 상층액을 취한 후 Griess 시약을 사용하여 제조사의 방법에 따라 NO 양을 측정하였다. LPS (400 ng / ml) was treated to RAW 274.7 cells divided into 48-well plates at an appropriate concentration, and the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (25, 50, 100, 200 µg / ml). Treated and incubated for 24 hours. After taking the culture supernatant, the amount of NO was measured according to the manufacturer's method using Griess reagent.
그 결과, 도 2에 개시한 바와 같이 LPS에 의해 증가된 NO 생성량이 석류풀 에탄올 추출물의 처리에 의해 농도 의존적으로 감소하였다. As a result, as shown in FIG. 2, the amount of NO produced increased by LPS was reduced in a concentration-dependent manner by treatment of pomegranate ethanol extract.
3) TNFα 측정3) TNFα measurement
48웰 플레이트에 적정 농도로 분주한 RAW 274.7 세포에 LPS(400ng/㎖)를 처리하고, 상기 실시예 1에서 제조한 석류풀 에탄올 추출물을 농도별(10, 50, 100, 200㎍/㎖)로 처리하여 24시간 동안 배양하였다. 배양 상층액을 취한 후 TNFα ELISA 키트를 사용하여 제조사의 방법에 따라 TNFα 생성량을 측정하였다. LPS (400 ng / ml) was treated to RAW 274.7 cells divided into 48-well plates at an appropriate concentration, and the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (10, 50, 100, 200 µg / ml). Treated and incubated for 24 hours. After taking the culture supernatant TNFα production was measured using the TNFα ELISA kit according to the manufacturer's method.
그 결과, 도 3에 개시한 바와 같이 LPS에 의해 증가된 TNFα 생성량이 석류풀 에탄올 추출물의 처리에 의해 96.9% 이상 감소하였다. As a result, as shown in FIG. 3, the amount of TNFα production increased by LPS was reduced by 96.9% or more by treatment of pomegranate ethanol extract.
4) PGE2(Prostaglandin E2) 측정4) Measurement of PGE 2 (Prostaglandin E 2 )
48웰 플레이트에 적정 농도로 분주한 RAW 274.7 세포에 LPS(400ng/㎖)를 처리하고, 상기 실시예 1에서 제조한 석류풀 에탄올 추출물을 농도별(25, 50, 100, 200㎍/㎖)로 처리하여 24시간 동안 배양하였다. 배양 상층액을 취한 후 PGE2 ELISA 키트를 사용하여 제조사의 방법에 따라 PGE2의 생성량을 측정하였다. LPS (400 ng / ml) was treated to RAW 274.7 cells divided into 48-well plates at an appropriate concentration, and the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (25, 50, 100, 200 µg / ml). Treated and incubated for 24 hours. After the culture supernatant was taken, the amount of PGE 2 was measured using the PGE 2 ELISA kit according to the manufacturer's method.
그 결과, 도 4에 개시한 바와 같이 LPS에 의해 증가된 PGE2 생성량이 40㎍/㎖의 석류풀 에탄올 추출물을 처리한 경우, 약 40% 정도 감소하였으며, 200㎍/㎖의 석류풀 에탄올 추출물을 처리한 경우는 99% 이상 PGE2 생성량이 감소하였다. As a result, as shown in FIG. 4, the amount of PGE 2 produced by LPS was decreased by about 40% when the 40 μg / ml pomegranate ethanol extract was treated, and the 200 μg / ml pomegranate ethanol extract was reduced. In the case of treatment, PGE 2 production decreased by more than 99%.
5) IL-6 측정5) IL-6 measurement
48웰 플레이트에 적정 농도로 분주한 RAW 274.7 세포에 LPS(400ng/㎖)를 처리하고, 상기 실시예 1에서 제조한 석류풀 에탄올 추출물을 농도별(25, 50, 100, 200㎍/㎖)로 처리하여 24시간 동안 배양하였다. 배양 상층액을 취한 후 IL-6 ELISA 키트를 사용하여 제조사의 방법에 따라 IL-6의 생성량을 측정하였다. LPS (400 ng / ml) was treated to RAW 274.7 cells divided into 48-well plates at an appropriate concentration, and the pomegranate ethanol extract prepared in Example 1 was prepared by concentration (25, 50, 100, 200 µg / ml). Treated and incubated for 24 hours. After taking the culture supernatant, the amount of IL-6 produced was measured according to the manufacturer's method using the IL-6 ELISA kit.
그 결과, 도 5에 개시한 바와 같이 LPS에 의해 증가된 IL-6 생성량이 석류풀 에탄올 추출물의 처리에 의해 농도 의존적으로 감소하였다. As a result, as shown in FIG. 5, the amount of IL-6 produced by LPS was decreased in a concentration-dependent manner by treatment of pomegranate ethanol extract.
실시예 3. MSU (Monosodium ureate )로 유도한 통풍성 관절염 동물모델에서 석류풀 추출물의 효능 평가 Example 3. The efficacy of seats ryupul extracts from gouty arthritis animal model induced by MSU (Monosodium ureate)
상기 실시예 1에서 추출한 석류풀 추출물이 MSU(Monosodium ureate)로 유도한 통풍성 관절염 질환 동물모델에서의 효능을 확인하기 위하여, 염증 사이토카인 IL-1β의 감소 효과를 확인하였다.  In order to confirm the efficacy of the pomegranate extract extracted in Example 1 in an animal model of gouty arthritis disease induced by MSU (Monosodium ureate), the effect of reducing the inflammatory cytokine IL-1β was confirmed.
상기 석류풀 메탄올 추출물, 석류풀 70%(v/v) 에탄올 추출물 및 석류풀 물 추출물을 각각 150, 300mg/kg 농도로 7주령 C57BL6 마우스에 경구 투여하였다. 상기 석류풀 추출물을 투여하고 1시간 후에 4mg의 MSU를 2.5% 트윈(tween) 80이 포함된 PBS 50㎕에 현탁하여 오른쪽 발조직(paw tissue)에 주사하여 통풍성 관절염을 유도하였다. 1일 1회 4일 동안 약물 투여한 후 MSU 유도 5일째에 부검하였다. The pomegranate grass methanol extract, pomegranate grass 70% (v / v) ethanol extract and pomegranate grass water extract were administered orally to 7-week-old C57BL6 mice at concentrations of 150 and 300 mg / kg, respectively. One hour after administration of the pomegranate extract, 4 mg of MSU was suspended in 50 μl of PBS containing 2.5% Tween 80 and injected into the right paw tissue to induce gouty arthritis. The drug was administered once a day for 4 days, followed by necropsy on the 5th day of MSU induction.
이후, 상기 부검에서 채취한 발 조직(paw tissue)을 RIPA 완충용액에 담그고 조직을 파쇄하여 상층액에서 염증성 지표인 IL-1β 단백질의 발현량을 ELISA법으로 측정하였고, 양성 대조군인 콜히친(cholchicine)을 사용하였다.Subsequently, the paw tissue obtained from the autopsy was immersed in RIPA buffer solution, and the tissue was crushed to measure the expression level of IL-1β protein, which is an inflammatory marker in the supernatant, by ELISA, and cholchicine as a positive control. Was used.
그 결과, 도 6에 개시한 바와 같이 통풍성 관절염으로 인해 증가된 IL-1β를 석류풀 추출물의 투여에 의해 IL-1β가 유의성 있게 감소하는 것을 확인하였다. As a result, as shown in FIG. 6, IL-1β increased due to gouty arthritis was significantly reduced by administration of pomegranate extract.
실시예Example 4. MIA로 유도한 골관절염 동물모델에서  4. In MIA-induced Osteoarthritis Animal Model 석류풀Pomegranate grass 추출물의 효능 평가 Evaluation of efficacy of extract
본 실시예 4에서는 상기 실시예 1에서 제조한 석류풀 70%(v/v) 에탄올 추출물이 MIA(monosodium iodoacetate)로 유도한 골관절염 질환 동물모델에서의 효능을 확인하였다. 7주령 SD 랫트에 석류풀 에탄올 추출물을 75, 150, 300mg/kg 농도로 3일 동안 경구 투여하였다. 약물을 투여한 후에 4일째(0 day)에 마취하고, SD 랫트의 오른쪽 무릎관절 주변을 깨끗이 제모한 후 골관절염 유발물질인 MIA를 관절강 내에 0.9% saline으로 희석하여 50㎕ (60mg/㎖)씩 투여하였다. 이후 1일 1회 14일 동안 석류풀 에탄올 추출물을 경구 투여한 후, 투여 14일째에 부검하였다. 양성대조군으로 인도메타신(indometacin)을 사용하였으며, 석류풀 추출물은 오전에 투여하고, 오후에 측정하였으며, 뒷다리 체중부하 측정일은 MIA 유발 후 -3, 0, 8, 11일째 되는 날에 각각 실시하였다.In Example 4, the pomegranate grass 70% (v / v) ethanol extract prepared in Example 1 was confirmed the efficacy in the osteoarthritis disease animal model induced by MIA (monosodium iodoacetate). Seven-week-old SD rats were orally administered with pomegranate ethanol extract at 75, 150, and 300 mg / kg concentrations for 3 days. Anesthesia was administered on day 4 (0 day) after administration of the drug, followed by clean hair removal around the right knee joint of the SD rat, and 50 μl (60 mg / ml) of MIA, a osteoarthritis-inducing substance, diluted with 0.9% saline in the joint cavity. It was. Thereafter, the pomegranate ethanol extract was orally administered once a day for 14 days, and then autopsied on the 14th day of administration. Indometacin was used as a positive control, and pomegranate extract was administered in the morning and measured in the afternoon, and hind limb weight measurement was performed on -3, 0, 8 and 11 days after MIA. .
1) 뒷다리 체중부하 측정 1) hind limb weight measurement
뒷다리 체중부하는 발 무게 측정기(Incapacitance tester, Linton instrument Co., UK, Ser No. 01/45/25)를 사용하여 측정하였다. 테스터의 홀더 안에서 골관절염이 유발된 랫트는 통증으로 인해 MIA를 투여하지 않은 정상적인 발에 의지하여 서게 되므로, 양쪽 발의 무게가 균형을 잃어 정상적인 발의 무게 대비 MIA를 투여한 발의 무게가 상대적으로 가볍게 측정되었다. 발의 무게 측정 시 SD 랫트의 배가 기기의 센서에 닿지 않은 상태에서 양쪽의 발의 무게(g)를 각각 측정하였다. Hind limb weight load was measured using a foot weigher (Incapacitance tester, Linton instrument Co., UK, Ser No. 01/45/25). In osteoarthritis-induced rats in the tester's holder, the pain of the osteoarthritis stood on the normal foot without MIA, so the weight of both feet was unbalanced and the weight of the MIA-treated foot was relatively light. When measuring the weight of the feet, the weight (g) of both feet was measured without the SD rat's belly touching the sensor of the device.
상기 측정된 발의 무게(g)를 이용하여, 체중부하율(%)을 계산하였다. 상기 체중부하는 발로 지탱하여 누르는 힘으로, 정상적인 경우 양발의 무게가 균형을 이루어 체중부하율(%)이 100%로 나타나지만, 통증이 심해질수록 체중부하율(%)이 낮아진다. Using the measured weight of the foot (g), the weight load rate (%) was calculated. The weight load is a force supported by the foot, the weight of both feet in the normal case, the weight load rate (%) appears as 100%, but the pain is worsened, the weight load rate (%) is lowered.
체중부하율(%) = (관절염이 유발된 뒷다리의 무게/정상 뒷다리의 무게)×100% Weight load = (weight of hind limb with arthritis / weight of normal hind limb) × 100
그 결과, 도 7에 개시한 바와 같이 MIA 유도에 의해 체중부하율이 현저하게 감소하였으나, MIA 유도 후 본 발명의 석류풀 추출물을 농도별로 투여한 경우 MIA 유도군에 비해 체중부하율(%)이 증가한 것을 확인할 수 있었다.As a result, as shown in FIG. 7, the weight-bearing rate was significantly reduced by MIA induction, but when the pomegranate extract of the present invention was administered by concentration after MIA induction, the weight-bearing rate (%) was increased compared to the MIA-induced group. I could confirm it.
2) 염증 사이토카인 ELISA 측정2) Inflammatory cytokine ELISA measurement
채혈한 혈액으로부터 혈청을 분리한 한 후, 혈청 중 염증성 지표로서 TNF-α 및 IL-6 단백질의 발현량을 ELISA법으로 측정하였다.After separating serum from the collected blood, expression levels of TNF-α and IL-6 protein as inflammatory markers in serum were measured by ELISA.
그 결과, 도 8에 개시한 바와 같이 MIA에 의해 증가된 TNF-α 및 IL-6 단백질의 발현량이 석류풀 추출물의 농도 의존적으로 감소한 것을 확인하였으며, TNF-α 단백질의 발현량은 150mg/kg 및 300mg/kg을 투여하였을 때 통계적으로 유의하게 감소하였으며, IL-6 단백질의 발현량은 300mg/kg을 투여하였을 때 통계적으로 유의하게 감소하였다.As a result, as shown in FIG. 8, it was confirmed that the expression level of TNF-α and IL-6 protein increased by MIA decreased in a concentration-dependent manner of pomegranate extract, and the expression level of TNF-α protein was 150 mg / kg and When 300 mg / kg was administered, it was statistically significantly decreased, and the expression level of IL-6 protein was statistically significantly decreased when 300 mg / kg was administered.

Claims (10)

  1. 석류풀(Mollugo pentaphylla) 추출물을 유효성분으로 함유하는 관절염의 예방 또는 개선용 건강기능식품 조성물.Pomegranate ( Mollugo pentaphylla ) Health functional food composition for the prevention or improvement of arthritis containing the extract as an active ingredient.
  2. 제1항에 있어서, 상기 관절염은 골 관절염 또는 통풍성 관절염인 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품 조성물.The health functional food composition for preventing or improving arthritis according to claim 1, wherein the arthritis is osteoarthritis or gouty arthritis.
  3. 제1항에 있어서, 상기 조성물은 염증성 사이토카인 및 통증을 억제하는 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품 조성물.The health functional food composition for preventing or improving arthritis according to claim 1, wherein the composition inhibits inflammatory cytokines and pain.
  4. 제3항에 있어서, 상기 통증은 관절염에 의한 통증인 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품 조성물.The health functional food composition for preventing or improving arthritis according to claim 3, wherein the pain is pain caused by arthritis.
  5. 제1항에 있어서, 상기 석류풀 추출물의 용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물인 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품 조성물.The health functional food composition for preventing or improving arthritis according to claim 1, wherein the solvent of the pomegranate extract is water, a lower alcohol of C 1 to C 4 or a mixture thereof.
  6. 제1항에 있어서, 상기 조성물은 분말, 과립, 환, 정제, 캡슐, 캔디, 시럽 및 음료 중에서 선택된 어느 하나의 제형으로 제조되는 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품 조성물.According to claim 1, wherein the composition is powder, granules, pills, tablets, capsules, candy, syrups and beverages, health functional food composition for the prevention or improvement of arthritis, characterized in that it is prepared in any one formulation.
  7. 석류풀 추출물을 유효성분으로 함유하는 관절염의 예방 또는 치료용 약학 조성물.Pharmaceutical composition for the prevention or treatment of arthritis containing pomegranate extract as an active ingredient.
  8. 제7항에 있어서, 상기 관절염은 골관절염 또는 통풍성 관절염인 것을 특징으로 하는 관절염의 예방 또는 치료용 약학 조성물.8. The pharmaceutical composition for preventing or treating arthritis according to claim 7, wherein the arthritis is osteoarthritis or gouty arthritis.
  9. 제7항에 있어서, 상기 조성물은 유효성분 이외에 약학적으로 허용 가능한 담체, 부형제 또는 희석제를 더 포함하는 것을 특징으로 하는 관절염의 예방 또는 치료용 약학 조성물.The pharmaceutical composition for preventing or treating arthritis of claim 7, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent in addition to the active ingredient.
  10. 제7항에 있어서, 상기 조성물은 캡슐제, 산제, 과립제, 정제, 현탁액, 에멀젼, 시럽, 에어로졸 중에서 선택된 어느 하나의 제형으로 제조되는 것을 특징으로 하는 관절염의 예방 또는 치료용 약학 조성물.According to claim 7, wherein the composition is a capsule, powder, granules, tablets, suspensions, emulsions, syrups, aerosol is a pharmaceutical composition for the prevention or treatment of arthritis, characterized in that prepared in any one of the formulations.
PCT/KR2017/011224 2016-10-26 2017-10-12 Composition for preventing, alleviating or treating arthritis, containing extract of mollugo pentaphylla as active ingredient WO2018080054A1 (en)

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