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WO2010016675A2 - Composition pharmaceutique de prevention et de traitement des maladies inflammatoires, contenant des extraits du fruit immature de rhus succedanea comme principes actifs - Google Patents

Composition pharmaceutique de prevention et de traitement des maladies inflammatoires, contenant des extraits du fruit immature de rhus succedanea comme principes actifs Download PDF

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WO2010016675A2
WO2010016675A2 PCT/KR2009/004187 KR2009004187W WO2010016675A2 WO 2010016675 A2 WO2010016675 A2 WO 2010016675A2 KR 2009004187 W KR2009004187 W KR 2009004187W WO 2010016675 A2 WO2010016675 A2 WO 2010016675A2
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Prior art keywords
inflammation
inflammatory
extract
present
inflammatory response
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Korean (ko)
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WO2010016675A3 (fr
Inventor
김환묵
강종순
박성규
이기호
이기훈
이창우
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Korea Research Institute of Bioscience and Biotechnology KRIBB
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Korea Research Institute of Bioscience and Biotechnology KRIBB
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    • 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)
    • A61K36/22—Anacardiaceae (Sumac family), e.g. smoketree, sumac or poison oak
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]

Definitions

  • the present invention relates to an anti-inflammatory composition and a method of treatment using the same.
  • Inflammation is the body's response to external invasive factors, such as infections, and protects the body by preventing external invasive factors from spreading.
  • excessive inflammation can cause a variety of diseases.
  • the development of many inflammation-related diseases is associated with the activation of macrophages and the excessive production of inflammation-related factors.
  • Representative inflammation-related factors include IL-1 ⁇ , TNF- ⁇ and nitrogen oxide (NO). They induce inflammation to remove external invaders, and when excessively produced, they destroy the tissues in vivo.
  • IL-1 ⁇ is produced mainly by macrophages activated with pro-inflammatory cytokines and is known to be involved in various immune and inflammatory responses.
  • NO is a free radical produced by various NOSs (eNOS, nNOS, iNOS, etc.) and is known to be involved in antibacterial, anticancer and vasodilating effects.
  • macrophages express large amounts of inducible nitric oxide synthase (iNOS), thus producing excess NO.
  • iNOS inducible nitric oxide synthase
  • Expression of IL-1 ⁇ and iNOS is regulated at the transcription level, and NF- ⁇ B is a transcription factor that plays an important role in the expression of these inflammatory mediators.
  • NF- ⁇ B is also known to play an important role in the expression of these inflammatory mediators as well as various immune and inflammatory processes.
  • Rhus succedanea is a dicotyledonous plant belonging to the Arachnoid family, which grows in low areas and is 13 m high. Leaves are alternate, about 30 cm long, 7-15 small leaves, long oval or elliptical basso pointed. Flowers are mixed with male and female. May bloom and yellow-green. Calyx, petals, and stamens are 5 each, and female flower has 5 small stamens, 1 ovary, and pistil is divided into 3 parts. Fruits are nucleus, flat, ball-shaped, yellow, without hairs, ripen in October, and harvested from the skin of fruit. It is distributed in Korea (Jeju Island, Jeonnam), Japan, China, Korea, and the Himalayas. It grows in the wild and is raised for ornamental purposes, but there is no record of medicinal use. Unlike the Rhus verniciflua, the lacquer does not rise.
  • the present inventors prepared the extract of the barley lacquer immature fruit, the extract is rapidly increased by inflammation, IL-1 ⁇ and NO production, IL-1 ⁇ and iNOS expression and NF- ⁇ B activity inhibition, dermatitis inhibition and death by sepsis
  • the present invention was completed by confirming that it can be used as a pharmaceutical composition for anti-inflammatory as a safe substance having excellent anti-inflammatory effect and no cytotoxicity.
  • An object of the present invention is to provide an anti-inflammatory treatment composition and a method for treating inflammation using the composition.
  • the present invention provides an anti-inflammatory composition containing an extract of Rhus succedanea immature.
  • the present invention provides an anti-inflammatory skin external preparation containing a barley lacquer immature fruit extract.
  • Another object of the present invention to provide a functional food for preventing or improving inflammation containing a barley lacquer immature fruit extract.
  • Another object of the present invention to provide a functional feed composition for preventing or improving inflammation containing barley lacquer immature fruit extract.
  • the present invention provides a method for treating inflammation using the composition.
  • Immature lacquer extract of the present invention is characterized by inhibiting NOx production, inhibiting the expression of inducible nitrogen oxide synthase (iNOS), and inhibiting and inhibiting the expression of interleukin-1 ⁇ (IL-1 ⁇ ), an proinflammatory cytokine.
  • iNOS inducible nitrogen oxide synthase
  • IL-1 ⁇ interleukin-1 ⁇
  • Immature lacquer extract of the present invention is characterized by inhibiting NOx production, inhibiting the expression of inducible nitrogen oxide synthase (iNOS), and inhibiting and inhibiting the expression of interleukin-1 ⁇ (IL-1 ⁇ ), an proinflammatory cytokine.
  • iNOS inducible nitrogen oxide synthase
  • IL-1 ⁇ interleukin-1 ⁇
  • Figure 1 is a view showing the results of confirming the cytotoxicity of the extract of the present invention, macrophages.
  • FIG. 2 is a diagram showing the inhibitory effect of IL-1 ⁇ and iNOS gene expression induced by LPS treatment in macrophages of the extract of the present invention.
  • FIG. 3 is a diagram showing the effect of inhibiting NF- ⁇ B activity induced by the treatment of LPS in macrophages of the extract of the present invention:
  • FIG. 4 is a diagram showing the dermatitis inhibitory effect of the extract of the present invention induced by the application of phorbol ester in BALB / c mice.
  • FIG. 5 is a diagram showing the inhibitory effect of death induced by LPS in BALB / c mice of the extract of the present invention.
  • the present invention provides a pharmaceutical composition for anti-inflammatory containing Rhus succedanea immature fruit extract as an active ingredient.
  • the present invention provides an anti-inflammatory skin external preparation containing the bark lacquer immature fruit extract as an active ingredient.
  • the present invention provides a functional food for preventing or improving inflammation containing a barley lacquer immature fruit extract as an active ingredient.
  • the present invention provides a functional feed composition for preventing or improving inflammation containing a barberry lacquer immature fruit extract as an active ingredient.
  • the present invention provides a method for treating inflammation comprising administering a pharmaceutically effective amount of immature lacquer ivy extract to an individual suffering from an inflammation-related disease.
  • anti-inflammatory refers to chronic inflammation that has progressed to a disease state, an inflammatory response generally caused by a harmless substance such as pollen, an inflammatory response caused by an inflammatory substance such as lipopolysaccharide (LPS), and asthma. It also refers to suppression of inflammatory reactions harmful to the human body, such as inflammatory reactions caused by autoimmune reactions such as rheumatoid arthritis.
  • the present invention provides an anti-inflammatory pharmaceutical composition containing Rhus succedanea immature fruit extract as an active ingredient.
  • It is preferably prepared by a manufacturing method comprising the step of concentrating the extract of step 2) under reduced pressure, but not limited to the above method.
  • the immature lacquer can be used without limitation, such as harvested, cultured or commercially available, in a preferred embodiment, it is preferable to use after drying to remove the foreign matter and salt by washing with water after harvesting.
  • the bark lacquer immature fruit extract may be prepared by conventional extraction methods in the art, such as ultrasonic extraction, filtration and reflux extraction.
  • it may be an extract extracted from the bark lacquer unripe and dried with water, a lower alcohol of C 1 ⁇ C 4 or a mixed solvent thereof, more preferably may be an extract extracted with a lower alcohol of C 1 ⁇ C 4 ,
  • the extract may be extracted with methanol or ethanol.
  • the immature lacquer tree and immature dry matter are ground to an appropriate size, put in an extraction container, methanol is extracted, and filtered using a sonicator, and then filtered with a filter paper to obtain a methanol extract.
  • the volume of the methanol solvent is preferably 5 to 15 times the volume of the ground sample.
  • the extraction time is preferably 1 to 5 days, most preferably 3 days. Thereafter, a method such as concentration or lyophilization may be further roughened.
  • the barley lacquer quince extract prepared by the above method inhibited IL-1 ⁇ and NOx (NO) production induced by LPS (lipopolysaccharide) treatment in a concentration-dependent manner (see Table 1).
  • LPS lipopolysaccharide
  • NF- ⁇ B plays an important role in gene expression of inflammatory mediators such as IL-1 ⁇ and iNOS.
  • the extract of the present invention suppressed NF- ⁇ B activity induced by LPS treatment in a concentration-dependent manner. (See Figure 3).
  • the dermatitis induced by phorbol ester by the extract of the present invention was concentration-dependently reduced (see FIG. 4), and it was confirmed that the death of sepsis mice induced by LPS was reduced ( See FIG. 5).
  • the extract of the present invention was shown to have no cytotoxicity at all, especially at a concentration of 0.1-3 ⁇ g / ml, inducing an anti-inflammatory effect (see FIG. 1).
  • the barley lacquer extract of the present invention is excellent in anti-inflammatory effect, it is judged to be very safe bar can be very useful in pharmaceutical compositions for anti-inflammatory.
  • the present invention contains 0.1 to 50% by weight of the barberry lacquer extract of the present invention with respect to the total weight of the pharmaceutical composition as an active ingredient, and may include a pharmaceutically acceptable carrier, excipient or diluent. Dosage forms of the pharmaceutical compositions of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.
  • the salt is not particularly limited as long as it is pharmaceutically acceptable, and for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid , Benzene sulfonic acid, toluene sulfonic acid, naphthalene sulfonic acid and the like can be used.
  • the pharmaceutical composition of the present invention may be in various oral or parenteral formulations.
  • diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
  • Solid form preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, which form 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, liquid solutions, emulsions, and syrups, and various excipients such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included.
  • Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories.
  • 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, glycerogelatin and the like can be used.
  • Inflammation may occur in a human body such as chronic inflammation progressing to a disease state, an inflammatory response generally caused by a harmless substance such as pollen, an inflammatory response caused by an inflammation-causing substance such as LPS, or an autoimmune response such as asthma or rheumatoid arthritis. Including but not limited to all harmful inflammatory reactions.
  • the subject to which the pharmaceutical composition of the present invention can be applied is a vertebrate, preferably a mammal, more preferably an experimental animal such as a rat, mouse, rabbit, guinea pig, hamster, dog, cat, and most preferably Are protozoan animals such as chimpanzees and gorillas.
  • the pharmaceutical composition of the present invention may be administered orally or parenterally, and when parenteral administration, it is preferable to select an external skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injection method. Most preferably, it is used for external skin.
  • the dosage of the pharmaceutical 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, the daily dosage is immature lacquer 0.1 to 100 mg / kg, preferably 30 to 80 mg / kg, more preferably 50 to 60 mg / kg, and can be administered 1 to 6 times per day based on the amount of the extract.
  • composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy and biological response modifiers.
  • the present invention provides an anti-inflammatory skin external preparation containing the bark lacquer immature fruit extract as an active ingredient.
  • the barley lacquer extract of the present invention is a concentration of IL-1 ⁇ and NO production induced by LPS treatment, the expression of the iNOS gene, the IL-1 ⁇ and NO producing enzyme and NF- ⁇ B activity Dependently (see Table 1 and Figures 2-3).
  • the dermatitis induced by phorbol ester by the extract of the present invention was concentration-dependently reduced (see FIG. 4), and it was confirmed that death of sepsis mice induced by LPS was reduced (see FIG. 5).
  • the extract of the present invention showed no cytotoxicity at all, especially at a concentration of 0.1-3 ⁇ g / ml, inducing anti-inflammatory effects (see FIG. 1).
  • the bark lacquer of the present invention immature fruit extract is excellent anti-inflammatory effect, it is determined that it is very safe, it can be used very useful in anti-inflammatory skin external preparations.
  • Inflammation may occur in a human body such as chronic inflammation progressing to a disease state, an inflammatory response generally caused by a harmless substance such as pollen, an inflammatory response caused by an inflammation-causing substance such as LPS, or an autoimmune response such as asthma or rheumatoid arthritis. Including but not limited to all harmful inflammatory reactions.
  • the barley lacquer extract of the present invention is additionally a fatty substance, an organic solvent, a dissolving agent, a thickening agent and a gelling agent, an emollient, an antioxidant, a suspending agent, a stabilizer, a foaming agent, a fragrance.
  • surfactants water, ionic or nonionic emulsifiers, fillers, metal ion sequestrants and chelating agents, preservatives, vitamins, blockers, wetting agents, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or skin It may contain adjuvants commonly used in the field of dermatology, such as any other ingredients commonly used in external preparations. In addition, the ingredients may be introduced in amounts generally used in the field of dermatology.
  • the present invention provides a functional food for preventing or improving inflammation containing a barley lacquer immature fruit extract as an active ingredient.
  • the barley lacquer extract of the present invention is a concentration of IL-1 ⁇ and NO production induced by LPS treatment, the expression of the iNOS gene, the IL-1 ⁇ and NO producing enzyme and NF- ⁇ B activity Dependently (see Table 1 and Figures 2-3).
  • the dermatitis induced by phorbol ester by the extract of the present invention was concentration-dependently reduced (see FIG. 4), and it was confirmed that death of sepsis mice induced by LPS was reduced (see FIG. 5).
  • the extract of the present invention showed no cytotoxicity at all, especially at a concentration of 0.1-3 ⁇ g / ml, inducing anti-inflammatory effects (see FIG. 1).
  • the bark lacquer of the present invention immature fruit extract is excellent anti-inflammatory effect, it is judged to be very safe bar, can be very useful for preventing or improving health functional foods.
  • Inflammation may occur in a human body such as chronic inflammation progressing to a disease state, an inflammatory response generally caused by a harmless substance such as pollen, an inflammatory response caused by an inflammation-causing substance such as LPS, or an autoimmune response such as asthma or rheumatoid arthritis. Including but not limited to all harmful inflammatory reactions.
  • the functional food according to the present invention may be, for example, various functional foods such as chewing gum, caramel products, candy, ice cream, confectionery, beverage products such as soft drinks, mineral water, alcoholic beverages, and health functional foods including vitamins and minerals. Can be.
  • the blending amount of the active ingredient may be appropriately determined depending on the purpose of use (prevention, health or hygiene).
  • the bark of the bark of the present invention is added in an amount of 15 parts by weight or less, preferably 10 parts by weight or less with respect to the raw material.
  • 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 functional food of the present invention may contain various flavors, natural carbohydrates, and the like as additional ingredients.
  • Natural carbohydrates described above are glucose, monosaccharides such as fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin, cyclodextrin, sugar alcohols such as 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 preferably selected in the range of 0.01 to 0.04 parts by weight, preferably about 0.02 to 0.03 parts by weight, per 100 parts by weight of the health food of the present invention.
  • the functional food 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 functional food of the present invention may contain a flesh for preparing natural fruit juice, fruit juice beverage and vegetable beverage. These components can be used independently or in combination. Although the ratio of such an additive is not critical, it is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health food of the present invention.
  • the present invention provides a functional feed composition for preventing or improving inflammation containing a barberry lacquer immature fruit extract as an active ingredient.
  • the barley lacquer extract of the present invention is a concentration of IL-1 ⁇ and NO production induced by LPS treatment, the expression of the iNOS gene, the IL-1 ⁇ and NO producing enzyme and NF- ⁇ B activity Dependently (see Table 1 and Figures 2-3).
  • the dermatitis induced by phorbol ester by the extract of the present invention was concentration-dependently reduced (see FIG. 4), and it was confirmed that death of sepsis mice induced by LPS was reduced (see FIG. 5).
  • the extract of the present invention showed no cytotoxicity at all, especially at a concentration of 0.1-3 ⁇ g / ml, inducing anti-inflammatory effects (see FIG. 1).
  • the bark lacquer of the present invention immature fruit extract is excellent anti-inflammatory effect, it is judged to be very safe, it can be very useful in the functional feed composition for preventing or improving inflammation.
  • the feed composition can be steadily ingested, thereby preventing inflammation and treating already occurring inflammation.
  • the present invention provides a method for treating inflammation comprising administering a pharmaceutically effective amount of immature lacquer ivy extract to an individual suffering from an inflammation-related disease.
  • the barley lacquer extract of the present invention is a concentration of IL-1 ⁇ and NO production induced by LPS treatment, the expression of the iNOS gene, the IL-1 ⁇ and NO producing enzyme and NF- ⁇ B activity Dependently (see Table 1 and Figures 2-3).
  • the bark immature fruit extract of the present invention inhibits IL-1 ⁇ and NO production, and effectively inhibits inflammation by inhibiting the expression of the IL-1 ⁇ and NO producing enzyme iNOS gene and NF- ⁇ B activity. It can be seen that.
  • the extract of the present invention showed no cytotoxicity at all, especially at a concentration of 0.1-3 ⁇ g / ml, which induces an anti-inflammatory effect (see FIG. 1).
  • the barley lacquer extract of the present invention can inhibit the effective inflammatory response in vivo.
  • the inflammation is also possible for inflammatory reactions other than those caused by LPS shown in the examples or inflammatory reactions with phorbol esters.
  • the extract of the present invention inhibits IL-1 ⁇ expression or inducible nitrogen oxide (NO) synthase (iNOS) expression, and thus is generally harmless, such as chronic inflammation and pollen, which can occur through this mechanism.
  • Harmful inflammatory reactions to the human body including inflammatory reactions by inflammatory reactions, inflammatory reactions by inflammatory agents such as LPS, and autoimmune reactions such as asthma or rheumatoid arthritis.
  • the subject to which the treatment method can be applied may be a vertebrate animal such as a mouse in addition to the mouse supported in the Examples. It is preferably a mammal, more preferably an experimental animal such as a rat, a mouse, a rabbit, a guinea peak, a hamster, a dog, a cat, and most preferably the same ape species such as chimpanzees and gorillas.
  • a vertebrate animal such as a mouse in addition to the mouse supported in the Examples. It is preferably a mammal, more preferably an experimental animal such as a rat, a mouse, a rabbit, a guinea peak, a hamster, a dog, a cat, and most preferably the same ape species such as chimpanzees and gorillas.
  • the treatment method in the embodiment shows the external skin and intraperitoneal injection method in the embodiment, but because there is no cytotoxicity, can be administered orally or parenterally in addition to the above method, rectal, vein, muscle, subcutaneous when parenteral administration Intrauterine epidural or cerebrovascular injection is preferred. Most preferably, it is used for external skin as in Example.
  • the dosage of the pharmaceutical composition in the treatment method may vary in the range depending on the weight, age, sex, health condition, administration time, administration method, excretion rate and severity of the patient.
  • Rhus succedanea obtained in Jeju Island was washed, dried completely using a dryer, and finely chopped.
  • the immature fruit of the finely chopped lacquer tree was immersed in 10 times of water and extracted by hot water for 3 days.
  • the extract was cooled, and the solvent of the supernatant obtained after filtration was evaporated in vacuo to concentrate the extract.
  • the yield was 5.1% (w / w).
  • the immature fruit of the finely chopped lacquer tree was immersed in 10% of 100% methanol (HPLC grade) and extracted using a sonicator (Branson Ultrasonics corporation, USA) for 3 days at room temperature. The extract was concentrated by evaporation in vacuo, yield 8.3% (w / w).
  • the extract was obtained by the method of Example 2 using 100% ethanol (HPLC grade). The yield was 7.4% (w / w).
  • FBS fetal bovine serum
  • penicillin 100 U / ml
  • streptomycin 100 ⁇ g / ml
  • BALB / c female mice of 18-22 g body weight (CORETECH, Gyeonggi-do) were cultured in the animal nursery under constant conditions (temperature: 21 ⁇ 2 ° C., contrast: 12 hour light / dark cycle). Free intake of feed and drinking water was allowed, and sufficient water and food were provided until the start of the experiment, and the animals were purified in the animal cage for 7 days.
  • the culture solution was removed, MTT reagent was added, and the reaction was performed at 37 ° C. incubator for 4 hours, and then MTT reagent was removed. After 100 ⁇ l DMSO was added, the cell viability was measured by measuring the absorbance at 540 nm using an ELISA reader.
  • IL-1 ⁇ production was performed using ELISA (Enzyme-linked immunosorbent assay) method using ELISA kit (USA) of R & D systems. It was measured by.
  • ELISA Enzyme-linked immunosorbent assay
  • Example 1 Example 2
  • Example 3 ( ⁇ g / ml) VH 0.1 0.3 One 3 VH 0.1 0.3 One 3 VH 0.1 0.3 One 3 IL-1 ⁇ production (ng /) 1.17 1.50 1.34 0.53 0.22 1.05 1.58 1.29 0.54 0.25 1.19 1.34 1.12 0.45 0.15 NO production (nmole / 10 6 cells) 39.6 43.9 42.2 34.7 18.0 41.7 42.8 42.8 37.4 14.2 42.2 42.2 42.2 34.7 19.6
  • the extract of the present invention inhibits the activation of macrophages.
  • RNA was treated with Experimental Example 1-2, after obtaining total RNA, IL-1 ⁇ and iNOS gene expression was measured using the reverse transcriptase-polymerase chain reaction (RT-PCR) method.
  • RT-PCR reverse transcriptase-polymerase chain reaction
  • the cells were recovered after the end of the culture and extracted with total RNA using RNeasy mini kit (QIAGEN, USA), and then 1 ⁇ g of total RNA and 1 ol of oligo-d (T) (Invitrogen, USA, 0.5 ⁇ g / ⁇ l) After filling up to 50 ⁇ l of distilled water, the reaction mixture was placed in AccuPower RT-premix (Bioneer, Korea) for RT-PCR reaction. 5 minutes at 70 ° C, 5 minutes at 4 ° C, 60 minutes at 42 ° C, 5 minutes at 94 ° C, and 5 minutes at 4 ° C to obtain cDNA.
  • RNeasy mini kit QIAGEN, USA
  • T oligo-d
  • PCR was performed using the primer pairs shown in Table 2 using the cDNA (3-5 ⁇ l) as a template.
  • cDNA 3-5 ⁇ l
  • PCR was performed. More specifically, denature for 5 minutes at 95 °C, turn 35 revolutions at the conditions of 95 °C 45 seconds, 65 °C 45 seconds, 72 °C 45 seconds, and then stretched for 5 minutes at 72 °C, then at 4 °C It was carried out in the order of cooling.
  • the PCR product generated after the completion of the reaction was electrophoresed on 1.5% agarose gel, and the size of the band was confirmed using a transilluminator.
  • PNF- ⁇ B-SEAP secreted alkaline phosphatase; Han MH et al. , Int Immunopharm , 7: 1651-1658, 2007
  • Lipofectamine Plus reagent Invitrogen, USA
  • the cells were transplanted into 96 well plates at a concentration of 5 ⁇ 10 5 cells per well, and then treated with LPS at a concentration of 300 ng / ml, and then the extracts were prepared at concentrations of 0.1, 0.3, 1 and 3 ⁇ g / ml. Treated.
  • NF- ⁇ B is known to play an important role in gene expression of inflammatory mediators such as IL-1 ⁇ and iNOS.
  • IL-1 ⁇ and iNOS inflammatory mediators
  • the inhibition of IL-1 ⁇ and iNOS gene expression by the extract of the present invention was found to be due to the inhibition of NF- ⁇ B activity.
  • the thickness of the ear was measured and the degree of edema was measured.
  • the thickness of the ear was measured using a digital thickness gauge (Digimatic Indicator, Japan).
  • mice prepared by the methods of Experiments 1-3 were intraperitoneally injected with 15 mg / kg LPS, 10 mg / kg of extract dissolved in phosphate buffered saline (PBS) added with 5% Tween 80, and PBS added with Tween 80. Injection.
  • PBS phosphate buffered saline
  • mice died within 2 days after LPS treatment, but in the extract treatment group of the present invention, about 70% of the mice died on day 2, and about 10% of mice after 7 days. Survived.
  • the extract of the present invention inhibits sepsis caused by LPS.
  • the present invention relates to an anti-inflammatory composition containing Rhus succedanea immature fruit extract as an active ingredient and a method for treating inflammation using the same, the immature lacquer fruit of the present invention exhibits an anti-inflammatory effect, and cytotoxicity is Therefore, it can be usefully used in medicines, nutraceuticals or functional feed for the prevention and treatment of inflammatory diseases.

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Abstract

L’invention concerne une composition anti-inflammatoire contenant des extraits du fruit immature de Rhus succedanea comme principes actifs. Lesdits extraits présentent des effets anti-inflammatoires par l’inhibition de la production d’oxyde d’azote (NO), l’inhibition de l’expression de la synthase d’oxyde nitrique inductible (iNOS) et l’inhibition de l’activité et de l’activité d’inhibition d’expression de l’interleukine-1β(IL-1β) qui est une cytokine pro-inflammatoire, et ils ne sont pas cytotoxiques. La présente invention peut donc être utilisée avantageusement dans des médicaments destinés à prévenir et à traiter les maladies inflammatoires, dans des aliments santé ou dans des aliments fonctionnels pour animaux.
PCT/KR2009/004187 2008-08-08 2009-07-28 Composition pharmaceutique de prevention et de traitement des maladies inflammatoires, contenant des extraits du fruit immature de rhus succedanea comme principes actifs Ceased WO2010016675A2 (fr)

Applications Claiming Priority (2)

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KR1020080078004A KR101009714B1 (ko) 2008-08-08 2008-08-08 검양옻나무 미숙과 추출물을 유효성분으로 포함하는 염증질환 예방 및 치료용 약학 조성물
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