KR20210050085A - Pharmaceutical composition comprising mixed extract of Gynostemma phetaphyllum and black radish - Google Patents
Pharmaceutical composition comprising mixed extract of Gynostemma phetaphyllum and black radish Download PDFInfo
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- KR20210050085A KR20210050085A KR1020190134238A KR20190134238A KR20210050085A KR 20210050085 A KR20210050085 A KR 20210050085A KR 1020190134238 A KR1020190134238 A KR 1020190134238A KR 20190134238 A KR20190134238 A KR 20190134238A KR 20210050085 A KR20210050085 A KR 20210050085A
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- extract
- mixed extract
- black radish
- dole
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Abstract
Description
본 발명은 돌외 및 검정무의 혼합추출물을 포함하는 약학 조성물에 관한 것으로, 보다 구체적으로 본 발명은 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만의 예방 또는 치료용 약학 조성물, 상기 약학 조성물을 이용하여 염증성 질환 또는 비만을 예방 또는 치료하는 방법, 상기 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만 개선용 식품조성물 및 상기 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만 개선용 사료조성물에 관한 것이다.The present invention relates to a pharmaceutical composition comprising a mixed extract of stone and black radish, and more specifically, the present invention, a pharmaceutical composition for the prevention or treatment of inflammatory disease or obesity, comprising a mixed extract of stone and black radish or a fraction thereof, A method for preventing or treating inflammatory disease or obesity using the pharmaceutical composition, a food composition for improving inflammatory disease or obesity, including the mixed extract of dolero and black radish or a fraction thereof, and a mixed extract of dolgeo and black radish or a mixture thereof It relates to a feed composition for improving inflammatory disease or obesity comprising a fraction.
일반적으로 염증반응은 세균감염과 같은 외부자극이나 생체 내 대사산물과 같은 내부자극에 대한 생체조직의 방어기전으로 세포 내 다양한 염증조절인자들인 TNF-α, IL-1β, IL-6 등과 같은 여러 사이토카인(cytokine) 및 산화질소(nitric oxide, NO)가 생성되면서 발생한다. 또한, 내독소로 알려진 지질다당체(lipopolysaccharide, LPS)는 그람 음성균의 세포 외막에 존재하여, 대식세포 또는 단핵세포에서 세포내 전사요소인 NF-κB(nuclear facter-κB)의 활성화를 유도함으로써 염증성 사이토카인, iNOS(inducible nitric oxide synthase), COX-2(cyclooxygenase-2)의 유전자 발현을 유도하며 염증 매개물질을 생성한다.In general, the inflammatory response is a defense mechanism of living tissues against external stimuli such as bacterial infection or internal stimuli such as metabolites in vivo. It occurs as cytokines and nitric oxide (NO) are produced. In addition, lipopolysaccharide (LPS), known as endotoxin, exists in the outer cell membrane of Gram-negative bacteria and induces the activation of NF-κB (nuclear facter-κB), an intracellular transcription factor, in macrophages or mononuclear cells. It induces gene expression of kine, iNOS (inducible nitric oxide synthase), and COX-2 (cyclooxygenase-2) and produces inflammatory mediators.
한편, 염증을 소실시키기 위해 염증원의 제거, 생체 반응 및 증상을 감소시키는 작용을 하는 것을 항염제라 한다. 현재까지 항염의 목적으로 이용되고 있는 물질로는 비스테로이드계로 플루폐나믹산(flufenamic acid), 이부프로펜(ibuprofen), 벤지다민(benzydamine), 인도메타신(indomethacin) 등이 있고 스테로이드계통으로 프레드니솔론(prednisolone), 덱사메타손 (dexamethasone) 등이 있다. 또한, 알란토인, 아즈엔, 하이드로코티손 등이 항염증에 효과가 있는 것으로 알려져 있으나, 이들 물질은 피부에 대한 안전성의 문제로 사용량의 제한이 있거나, 효과가 미미하여 실질적으로 염증 완화 효과를 기대할 수 없는 문제점이 있다.On the other hand, in order to eliminate inflammation, it is called an anti-inflammatory agent that acts to remove inflammatory sources, reduce biological reactions, and symptoms. Substances used for anti-inflammatory purposes to date include flufenamic acid, ibuprofen, benzydamine, indomethacin, etc. as non-steroids, and prednisolone as a steroidal system , Dexamethasone, and the like. In addition, allantoin, azene, hydrocortisone, etc. are known to have anti-inflammatory effects, but these substances are limited in use due to safety issues on the skin, or the effect is insignificant, so the effect of substantially reducing inflammation cannot be expected. There is this.
이러한 배경하에서, 본 발명자들은 보다 안전하면서도 효과적으로 염증성 질환을 치료하는 방법을 개발하고자 예의 연구노력한 결과, 돌외와 검정무의 혼합추출물이 안전하면서도 효과적으로 염증성 질환을 치료하는 효과를 나타낼 뿐만 아니라, 비만증상 까지도 치료하는 효과를 나타냄을 확인하고, 본 발명을 완성하였다.Under this background, the present inventors have made intensive research efforts to develop a safer and more effective method for treating inflammatory diseases. As a result, the mixed extract of stone and black radish not only safely and effectively treats inflammatory diseases, but also obesity symptoms. It was confirmed that the treatment effect was shown, and the present invention was completed.
본 발명의 하나의 목적은 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만의 예방 또는 치료용 약학 조성물을 제공하는 것이다.One object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases or obesity, including a mixed extract or fractions thereof of oleum and black radish.
본 발명의 다른 목적은 상기 약학 조성물을 이용하여 염증성 질환 또는 비만을 예방 또는 치료하는 방법을 제공하는 것이다.Another object of the present invention is to provide a method for preventing or treating inflammatory diseases or obesity using the pharmaceutical composition.
본 발명의 또 다른 목적은 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만 개선용 식품조성물을 제공하는 것이다.Another object of the present invention is to provide a food composition for improving inflammatory disease or obesity, including a mixed extract of dole and black radish or a fraction thereof.
본 발명의 또 다른 목적은 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만 개선용 사료조성물을 제공하는 것이다.Another object of the present invention is to provide a feed composition for improving inflammatory disease or obesity, including a mixed extract of dole and black radish or a fraction thereof.
상술한 목적을 달성하기 위한 본 발명의 일 실시양태는 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 염증성 질환 또는 비만의 예방 또는 치료용 약학 조성물을 제공한다.One embodiment of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of inflammatory diseases or obesity, including a mixed extract of extradolescence and black radish or a fraction thereof.
본 발명의 용어 "돌외(Gynostemma phetaphyllum)"란, 박과(Cucurbitaceae)에 속하고 제주 전역에 자생하는 덩굴성 식물을 의미한다. 상기 돌외는 지페노사이드, 플라보노이드, 폴리사카라이드 등의 성분을 다량으로 포함하여, 면역조절 활성, 항산화 활성, 항암 활성, 항고지혈증 활성, 신경세포보호 활성, 간세포보호 활성 등을 나타낸다고 알려져 있다. 다만, 돌외는 생체에 독성을 유발하는 것으로 알려져 있어, 현재로서는 돌외 잎을 덖음 가공하여 차의 형태로 이용하고 있다.The term "Gynostemma phetaphyllum" of the present invention means a vine-like plant belonging to the Cucurbitaceae family and growing all over Jeju. It is known that the dolores contain a large amount of components such as zipenosides, flavonoids, and polysaccharides, and exhibit immunomodulatory activity, antioxidant activity, anticancer activity, antihyperlipidemia activity, neuroprotective activity, hepatocyte protective activity, and the like. However, as it is known to cause toxicity to the living body, it is currently used in the form of tea by grinding and processing the leaves of the stone.
본 발명의 용어 "검정무(black radish; Raphanus sativus L. Var niger)"란, 십자과(Brassicaceae)에 속하는 지중해 연안이 원산지인 채소류를 의미한다. 상기 검정무에는 글루코시놀레이트(glucosinolate)가 다량으로 포함되어 있어, 위장질환, 담낭질환, 간질환 등의 질환에 대한 치료효과를 나타낸다고 알려져 있다.The term "black radish (Raphanus sativus L. Var niger)" of the present invention refers to vegetables native to the Mediterranean coast belonging to the cruciferous family (Brassicaceae). Since the black radish contains a large amount of glucosinolate, it is known that it exhibits a therapeutic effect on diseases such as gastrointestinal disease, gallbladder disease, and liver disease.
본 발명의 용어 "추출물"이란, 목적하는 물질을 다양한 용매에 침지한 다음, 상온 또는 가온상태에서 일정시간 동안 추출하여 수득한 액상성분, 상기 액상성분으로부터 용매를 제거하여 수득한 고형분 등의 결과물을 의미한다. 뿐만 아니라, 상기 결과물에 더하여, 상기 결과물의 희석액, 이들의 농축액, 이들의 조정제물, 정제물 등을 모두 포함하는 것으로 포괄적으로 해석될 수 있다. 이에 따라, 본 발명에서 제공하는 돌외 및 검정무의 혼합추출물은 돌외 및 검정무를 추출 처리하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함하는 것으로 해석될 수 있다. The term "extract" of the present invention refers to a liquid component obtained by immersing a target substance in various solvents and then extracting it for a certain period of time at room temperature or warming state, and a resultant product such as a solid component obtained by removing the solvent from the liquid component. it means. In addition, in addition to the resulting product, it can be comprehensively interpreted as including all of the diluted solution of the resultant, the concentrated solution thereof, the preparation thereof, and the purified product. Accordingly, the mixed extract of boulder and black radish provided in the present invention is an extract obtained by extracting boulder and black radish, a dilution or concentrate of the extract, a dried product obtained by drying the extract, a prepared product or purified product of the extract, Or it may be interpreted as including a mixture thereof, the extract itself and the extract of all formulations that can be formed using the extract.
본 발명의 돌외 및 검정무의 혼합추출물에 있어서, 상기 혼합물을 추출하는 방법은 특별히 제한되지 않으며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출 방법의 비제한적인 예로는, 열수 추출법, 초음파 추출법, 여과법, 환류 추출법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2종 이상의 방법을 병용하여 수행될 수 있다.In the mixed extract of dolgae and black radish of the present invention, a method of extracting the mixture is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include a hot water extraction method, an ultrasonic extraction method, a filtration method, a reflux extraction method, and the like, which may be performed alone or in combination of two or more methods.
본 발명에서 상기 추출에 사용되는 용매의 종류는 특별히 제한되지 않으며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 추출 용매의 비제한적인 예로는 물, 알코올 또는 이들의 혼합 용매 등을 들 수 있고, 이들은 단독으로 사용되거나 1종 이상 혼합하여 사용될 수 있으며, 구체적으로 물이 사용될 수 있다. 알코올을 용매로 사용하는 경우에는 구체적으로 탄소수 1 내지 4의 알코올을 사용할 수 있다.In the present invention, the type of solvent used for the extraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the extraction solvent may include water, alcohol, or a mixed solvent thereof, and these may be used alone or in combination of one or more, and specifically water may be used. When alcohol is used as a solvent, an alcohol having 1 to 4 carbon atoms may be specifically used.
본 발명에서 제공하는 돌외 및 검정무의 혼합추출물은 돌외 추출물과 검정무 추출물을 혼합하여 제조하거나 또는 돌외와 검정무의 혼합물을 추출하여 수득할 수 있는데, 상기 혼합 추출물은 돌외 추출물이 나타내는 세포독성을 완화시킬 수 있으면서도, 항염증 효과 또는 항비만 효과를 나타낼 수 있도록, 일 예로서, 돌외 추출물과 검정무 추출물을 1:5 내지 1:9(w/w)로 혼합한 것이 될 수 있고, 다른 예로서, 돌외 추출물과 검정무 추출물을 1:7 내지 1:9(w/w)로 혼합한 것이 될 수 있으며, 또 다른 예로서 돌외 추출물과 검정무 추출물을 1:9(w/w)로 혼합한 것이 될 수 있다. The mixed extract of dole and black radish provided by the present invention may be prepared by mixing a dole extract and a black radish extract, or may be obtained by extracting a mixture of a dole and black radish, the mixed extract exhibiting the cytotoxicity of the dole extract. As an example, it may be a mixture of 1 :5 to 1:9 (w/w) of a stone extract and a black radish extract so as to be able to alleviate it and exhibit an anti-inflammatory effect or an anti-obesity effect, and another example As, it may be a mixture of the Dole extract and the black radish extract 1:7 to 1:9 (w/w), and as another example, the Dole extract and the black radish extract are mixed at 1:9 (w/w). It can be one.
상기 혼합 추출물은 염증성 질환에 관여하는 다양한 염증인자인 NO, Cox-2, iNOS, p65, JNK 등의 발현을 억제하는 효과를 나타낼 수 있다. 뿐만 아니라, 상기 혼합 추출물은 지방세포의 분화를 억제하고, 지방세포내 중성지방의 축적을 억제하며, 지방세포 분화인자 PPARγ 및 aP2의 발현을 억제하는 효과를 나타낼 수 있다. The mixed extract may exhibit an effect of inhibiting the expression of various inflammatory factors involved in inflammatory diseases such as NO, Cox-2, iNOS, p65, and JNK. In addition, the mixed extract may exhibit an effect of inhibiting the differentiation of adipocytes, inhibiting the accumulation of triglycerides in adipocytes, and inhibiting the expression of adipocyte differentiation factors PPARγ and aP2.
본 발명의 용어 "분획물"이란, 여러 다양한 구성 성분들을 포함하는 혼합물로부터 특정 성분 또는 특정 성분 그룹을 분리하기 위하여 분획을 수행하여 얻어진 결과물을 의미한다.The term "fraction" in the present invention means a result obtained by performing fractionation in order to separate a specific component or a specific group of components from a mixture containing several different constituents.
본 발명에서 상기 분획물을 얻는 분획 방법은 특별히 제한되지 않으며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 수행될 수 있다. 상기 분획 방법의 비제한적인 예로는, 다양한 용매를 처리하여 수행하는 용매 분획법, 일정한 분자량 컷-오프 값을 갖는 한외 여과막을 통과시켜 수행하는 한외여과 분획법, 다양한 크로마토그래피(크기, 전하, 소수성 또는 친화성에 따른 분리를 위해 제작된 것)를 수행하는 크로마토그래피 분획법, 및 이들의 조합 등이 있다. 구체적으로, 본 발명의 돌외 및 검정무의 혼합추출물에 소정의 용매를 처리하여 상기 추출물로부터 분획물을 얻는 방법을 들 수 있다.The fractionation method for obtaining the fraction in the present invention is not particularly limited, and may be performed according to a method commonly used in the art. Non-limiting examples of the fractionation method include a solvent fractionation method performed by treating various solvents, an ultrafiltration fractionation method performed by passing an ultrafiltration membrane having a certain molecular weight cut-off value, and various chromatography (size, charge, hydrophobicity). Or a chromatographic fractionation method for performing separation according to affinity), and a combination thereof. Specifically, there may be mentioned a method of obtaining a fraction from the extract by treating the mixed extract of dole and black radish of the present invention with a predetermined solvent.
본 발명에서 상기 분획물을 얻는 데 사용되는 분획 용매의 종류는 특별히 제한되지 않으며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 분획 용매의 비제한적인 예로는 물, 탄소수 1 내지 4의 알코올 등의 극성 용매; 헥산(Hexan), 에틸 아세테이트(Ethyl acetate), 클로로포름(Chloroform), 디클로로메탄(Dichloromethane) 등의 비극성 용매; 또는 이들의 혼합용매 등을 들 수 있다. 이들은 단독으로 사용되거나 1종 이상 혼합하여 사용될 수 있지만, 이에 제한되는 것은 아니다.In the present invention, the type of fractionation solvent used to obtain the fraction is not particularly limited, and any solvent known in the art may be used. Non-limiting examples of the fractionation solvent include polar solvents such as water and alcohols having 1 to 4 carbon atoms; Non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane; Or a mixed solvent of these, etc. are mentioned. These may be used alone or in combination of one or more, but are not limited thereto.
또한, 상기 추출물 또는 분획물은 추출 후 건조 분말 형태로 제조되어 사용될 수 있지만, 이제 제한되는 것은 아니다.In addition, the extract or fraction may be prepared and used in a dry powder form after extraction, but is not limited thereto.
본 발명의 용어 "염증성 질환(Inflammatory disease)"이란, 염증유발인자 또는 방사선조사 등 유해한 자극으로 인해 인체 면역체계를 과도하게 항진시켜 대식세포와 같은 면역세포에서 분비되는 TNF-α(tumor necrosis factor-α), IL-1(interleukin-1), IL-6, 프로스타글란딘(prostagladin), 루코트리엔(luecotriene) 또는 산화질소(nitric oxide, NO)와 같은 염증유발물질(염증성 사이토카인)에 의해 유발되는 질환을 의미한다. The term "inflammatory disease" of the present invention refers to TNF-α (tumor necrosis factor-) secreted by immune cells such as macrophages by excessively stimulating the human immune system due to harmful stimulation such as inflammation inducing factors or irradiation. α), IL-1 (interleukin-1), IL-6, prostaglandin (prostagladin), leukotriene (luecotriene) or nitric oxide (nitric oxide, NO) such as pro-inflammatory substances (inflammatory cytokines) It means a disease that becomes.
본 발명에 있어서, 상기 염증성 질환은 특별히 이에 제한되지 않으나, 일 예로서, 알레르기성 염증질환, 염증성 피부질환, 염증성 안구질환, 염증성 골관절질환, 염증성 근육질환, 염증성 장질환 등이 될 수 있고, 다른 예로서, 알레르기 비염, 맥관부종, 알러지성 결막염 등의 알레르기성 염증질환; 아토피피부염, 건선, 접촉성피부염, 습진성 피부염, 광선피부염, 지루피부염, 포진성피부염, 편평태선, 경화태선, 괴저성 농피증, 천포창, 수포성 표피박리증 등의 염증성 피부질환; 안검염, 퇴행성 또는 염증성 안구염 등의 염증성 안구질환; 관절염, 류마티스관절염, 척추염 등의 염증성 골관절질환; 전신성 경화증, 피부근염, 다발성근염, 염증성 근병변 등의 염증성 근육질환; 궤양성 대장염, 크론병 등의 염증성 장질환이 될 수 있다.In the present invention, the inflammatory disease is not particularly limited thereto, but as an example, it may be an allergic inflammatory disease, inflammatory skin disease, inflammatory eye disease, inflammatory bone and joint disease, inflammatory muscle disease, inflammatory bowel disease, and the like. For example, allergic inflammatory diseases such as allergic rhinitis, angioedema, and allergic conjunctivitis; Inflammatory skin diseases such as atopic dermatitis, psoriasis, contact dermatitis, eczema dermatitis, photodermatitis, seborrheic dermatitis, herpes dermatitis, lichen planus, lichen sclerosis, gangrene pyoderma, pemphigus, and vesicular epidermolysis; Inflammatory eye diseases such as blepharitis, degenerative or inflammatory ophthalmitis; Inflammatory bone and joint diseases such as arthritis, rheumatoid arthritis, and spondylitis; Inflammatory muscle diseases such as systemic sclerosis, dermatomyositis, polymyositis, and inflammatory muscle lesions; It can be an inflammatory bowel disease such as ulcerative colitis and Crohn's disease.
본 발명의 용어 "비만"이란, 일반적으로 체내에 지방조직이 과다한 상태이거나 또는 신체비만지수(체질량지수, Body mass index: 체중(kg)을 신장(m)의 제곱으로 나눈 값)가 25 이상인 경우를 의미한다. 비만증상이 발생한 개체에서는 통상적으로 혈장으로부터 지방세포로 유입된 지방산과 포도당이 에스테르화하여 주로 중성지방의 형태로 축적된다. 본 발명의 목적상 상기 비만증상은 본 발명의 조성물의 투여로 인하여 예방, 경감, 개선 또는 치료될 수 있는 목적질환으로 사용되지만, 특별히 이에 제한되지는 않는다.The term "obesity" in the present invention generally refers to a state in which adipose tissue is excessive in the body, or when the body obesity index (body mass index: the value obtained by dividing the body weight (kg) by the square of the height (m)) is 25 or more. Means. In individuals with obesity symptoms, fatty acids and glucose, which are introduced from plasma into adipocytes, are esterified and accumulated mainly in the form of triglycerides. For the purposes of the present invention, the obesity symptom is used as a target disease that can be prevented, alleviated, improved or treated by administration of the composition of the present invention, but is not particularly limited thereto.
본 발명의 용어 "예방"이란, 상기 약학 조성물의 투여로 염증성 질환 또는 비만을 억제 또는 지연시키는 모든 행위를 의미한다. The term "prevention" of the present invention means any action of inhibiting or delaying inflammatory disease or obesity by administration of the pharmaceutical composition.
본 발명의 용어 "치료"란, 치료하고자 하는 개개인 또는 세포의 천연 과정을 변경시키기 위해 임상적으로 개입하는 모든 행위를 의미하는데, 임상 병리 상태가 진행되는 동안 또는 이를 예방하기 위해 수행할 수 있다. 목적하는 치료 효과에는 질병의 발생 또는 재발을 예방하고, 증상을 완화시키며, 질병에 따른 모든 직접 또는 간접적인 병리학적 결과를 저하시키며, 전이를 예방하고, 질병 진행 속도를 감소시키며, 질병 상태를 경감 또는 일시적 완화시키며, 차도시키거나 예후를 개선시키는 것이 포함된다. 본 발명의 목적상 상기 치료는 상기 약학 조성물의 투여로 염증성 질환 또는 비만의 증세가 호전되거나 완치되는 모든 행위를 포함하는 것으로 해석될 수 있으나, 특별히 이에 제한되지는 않는다.The term "treatment" of the present invention refers to any action that clinically intervenes in order to alter the natural process of an individual or cell to be treated, and can be performed while a clinical pathology is progressing or to prevent it. The desired therapeutic effect includes preventing the occurrence or recurrence of the disease, alleviating symptoms, reducing all direct or indirect pathological consequences of the disease, preventing metastasis, reducing the rate of disease progression, and alleviating the disease state. Or temporary alleviation, remission or improving prognosis. For the purposes of the present invention, the treatment may be interpreted as including all actions in which symptoms of inflammatory disease or obesity are improved or cured by administration of the pharmaceutical composition, but is not particularly limited thereto.
상기 약학 조성물은 약학 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 추가로 포함할 수 있는데, 상기 담체는 비자연적인 담체가 될 수 있다. 상기 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. The pharmaceutical composition may further include an appropriate carrier, excipient, and diluent commonly used in the preparation of pharmaceutical compositions, and the carrier may be an unnatural carrier. The carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oils.
한편, 본 발명의 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제제화하여 사용될 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 돌외 및 검정무의 혼합추출물 또는 그의 분획물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트 (calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스티레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는 데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제가 포함된다. 비수성용제, 현탁제로는 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, 글리세로젤라틴 등이 사용될 수 있다. Meanwhile, the pharmaceutical composition of the present invention is formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, and sterile injectable solutions, respectively, according to conventional methods. I can. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient, such as starch, calcium carbonate, in the mixed extract or fraction thereof It is prepared by mixing (calcium carbonate), sucrose or lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral use include water and liquid paraffin, which are simple diluents commonly used for suspensions, liquid solutions, emulsions, syrups, etc., and various excipients such as wetting agents, sweetening agents, fragrances, and preservatives. have. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol,
상기 약학적 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁제, 내용액제, 유제, 시럽제, 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제 및 좌제로 이루어진 군으로부터 선택되는 어느 하나의 제형을 가질 수 있다.The pharmaceutical composition is any selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, solutions, emulsions, syrups, sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. It can have one formulation.
본 발명의 상기 약학 조성물에 포함된 돌외 및 검정무의 혼합추출물 또는 그의 분획물의 함량은 특별히 이에 제한되지 않으나, 최종 조성물 총 중량에 대하여 0.0001 내지 50 중량%로 포함할 수 있고, 바람직하게는 0.001 내지 10 중량%의 함량으로 포함할 수 있다. The content of the mixed extract of dole and black radish or fractions thereof included in the pharmaceutical composition of the present invention is not particularly limited thereto, but may be included in an amount of 0.0001 to 50% by weight based on the total weight of the final composition, and preferably 0.001 to It may be included in an amount of 10% by weight.
상기 본 발명의 약학 조성물은 약학적으로 유효한 양으로 투여될 수 있는데, 본 발명의 용어 "약학적으로 유효한 양"이란, 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 개체 종류 및 중증도, 연령, 성별, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 약학 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있다. 그리고 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하다.The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount, and the term "pharmaceutically effective amount" of the present invention means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment. The effective dose level is well suited to the type and severity of the subject, age, sex, activity of the drug, sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including concurrent drugs, and other medical fields. It can be determined according to known factors. The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered single or multiple. Considering all of the above factors, it is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects.
본 발명의 약학 조성물의 투여량은 사용목적, 질환의 중독도, 환자의 연령, 체중, 성별, 기왕력, 또는 유효성분으로서 사용되는 물질의 종류 등을 고려하여 당업자가 결정할 수 있다. 예를 들어, 본 발명의 약학 조성물은 성인 1인당 약 0.1 ng 내지 약 100 mg/kg, 바람직하게는 1 ng 내지 약 10 mg/kg로 투여할 수 있고, 본 발명의 조성물의 투여빈도는 특별히 이에 제한되지 않으나, 1일 1회 투여하거나 또는 용량을 분할하여 수회 투여할 수 있다.The dosage of the pharmaceutical composition of the present invention can be determined by a person skilled in the art in consideration of the purpose of use, the degree of addiction to the disease, the age, weight, sex, history, or type of substance used as an active ingredient. For example, the pharmaceutical composition of the present invention can be administered in an amount of about 0.1 ng to about 100 mg/kg, preferably 1 ng to about 10 mg/kg per adult, and the frequency of administration of the composition of the present invention is specifically Although not limited, it may be administered once a day or divided into several doses.
다른 양태로서, 본 발명은 상기 약학 조성물을 염증성 질환 또는 비만이 발병되거나 또는 발병될 가능성이 있는, 인간을 제외한 개체에 투여하는 단계를 포함하는 염증성 질환 또는 비만의 예방 또는 치료방법을 제공한다.In another aspect, the present invention provides a method for preventing or treating an inflammatory disease or obesity, comprising administering the pharmaceutical composition to an individual other than humans, which has or is likely to develop an inflammatory disease or obesity.
본 발명의 용어 "개체"란, 상기 염증성 질환 또는 비만이 발병될 가능성이 있거나, 또는 발병된 조직을 포함하는 모든 동물을 의미하는데, 인간은 제외된다. 본 발명의 조성물을 개체에 투여함으로써, 염증성 질환 또는 비만을 완화 또는 치료할 수 있다. The term "individual" of the present invention refers to all animals including tissues that are likely to develop the inflammatory disease or obesity, or have the disease, except humans. By administering the composition of the present invention to an individual, inflammatory disease or obesity can be alleviated or treated.
본 발명의 용어, "완화"는 본 발명에 따른 조성물의 투여로 염증성 질환 또는 비만이 호전되거나 이롭게 되는 모든 행위를 말한다.The term "relieving" of the present invention refers to any action in which an inflammatory disease or obesity is ameliorated or benefited by administration of the composition according to the present invention.
본 발명의 용어 "치료"란, 본 발명에 따른 조성물을 염증성 질환 또는 비만의 치료가 요구되는 개체에 투여하여 염증성 질환 또는 비만의 치료가 수행되거나 이롭게 되도록 하는 모든 행위를 의미한다.The term "treatment" of the present invention refers to any action in which the composition according to the present invention is administered to an individual in need of treatment of an inflammatory disease or obesity to perform or benefit from the treatment of an inflammatory disease or obesity.
상기 본 발명의 조성물은 약학적으로 유효한 양으로 투여한다.The composition of the present invention is administered in a pharmaceutically effective amount.
본 발명에서 용어, "투여"는 어떠한 적절한 방법으로 대상에게 본 발명의 약학적 조성물을 도입하는 것을 말하며, 투여 경로는 목적 조직에 도달할 수 있는 한 경구 또는 비경구의 다양한 경로를 통하여 투여될 수 있다.In the present invention, the term "administration" refers to introducing the pharmaceutical composition of the present invention to a subject by any suitable method, and the route of administration may be administered through various routes, either oral or parenteral, as long as it can reach the target tissue. .
상기 약학적 조성물은 목적 또는 필요에 따라 당업계에서 사용되는 통상적인 방법, 투여 경로, 투여량에 따라 적절하게 개체에 투여될 수 있다. 투여 경로의 예로는 경구, 비경구, 피하, 복강 내, 폐 내, 및 비강 내로 투여될 수 있으며, 비경구 주입에는 근육 내, 정맥 내, 동맥 내, 복강 내 또는 피하투여가 포함된다. 또한 당업계에 공지된 방법에 따라 적절한 투여량 및 투여 횟수가 선택될 수 있으며, 실제로 투여되는 본 발명의 약학적 조성물의 양 및 투여 횟수는 치료하고자 하는 증상의 종류, 투여 경로, 성별, 건강 상태, 식이, 개체의 연령 및 체중, 및 질환의 중증도와 같은 다양한 인자에 의해 적절하게 결정될 수 있다.The pharmaceutical composition may be appropriately administered to an individual according to a conventional method, route of administration, and dosage used in the art, depending on the purpose or need. Examples of the route of administration may be oral, parenteral, subcutaneous, intraperitoneal, intrapulmonary, and intranasal administration, and parenteral infusion includes intramuscular, intravenous, intraarterial, intraperitoneal or subcutaneous administration. In addition, an appropriate dosage and number of administrations may be selected according to methods known in the art, and the amount and number of administrations of the pharmaceutical composition of the present invention actually administered can be determined by the type of symptom to be treated, route of administration, sex, health condition , Diet, the age and weight of the individual, and the severity of the disease.
본 발명에서의 용어 "약학적으로 유효한 양"은 의학적 용도에 적용 가능한 합리적인 수혜/위험 비율로 혈관 투과성 증가를 억제 또는 완화하기에 충분한 양을 의미하며, 유효 용량 수준은 개체 종류 및 중증도, 연령, 성별, 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 있다. 그리고 단일 또는 다중 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 당업자에 의해 용이하게 결정될 수 있다.The term "pharmaceutically effective amount" as used herein refers to an amount sufficient to inhibit or alleviate the increase in vascular permeability at a reasonable benefit/risk ratio applicable to medical use, and the effective dose level is the type and severity of the subject, age, Sex, activity of the drug, sensitivity to the drug, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, and may be administered sequentially or simultaneously with a conventional therapeutic agent. And can be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and can be easily determined by a person skilled in the art.
또 다른 양태로서, 본 발명은 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는, 염증성 질환 또는 비만 개선용 식품 조성물을 제공한다.In another aspect, the present invention provides a food composition for improving inflammatory diseases or obesity, including a mixed extract of dole and black radish or a fraction thereof.
본 발명의 용어, "식품"은 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올음료, 비타민 복합제, 건강 기능 식품 및 건강 식품 등이 있으며, 통상적인 의미에서의 식품을 모두 포함한다.As used herein, the term "food" refers to meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages , Vitamin complexes, health functional foods, and health foods, and all foods in the usual sense are included.
본 발명의 식품 조성물은 일상적으로 섭취가능한 돌외 및 검정무로부터 유래되었기 때문에 높은 염증성 질환 또는 비만 개선 효과를 기대할 수 있으므로, 건강 증진 목적으로 매우 유용하게 사용될 수 있다.Since the food composition of the present invention is derived from daily intakeable dole and black radish, it can be expected to have a high inflammatory disease or obesity improvement effect, and thus can be very usefully used for health promotion purposes.
상기 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 조성물은 식품 조성물의 총 중량에 대하여 0.01 내지 100 중량%, 보다 바람직하게는 1 내지 80 중량%로 포함한다. 식품이 음료인 경우에는 100㎖를 기준으로 1~30g, 바람직하게는 3 ~ 20g의 비율로 포함될 수 있다. 또한, 상기 조성물은 식품 조성물에 통상 사용되어 냄새, 맛, 시각 등을 향상시킬 수 있는 추가 성분을 포함할 수 있다. 예들 들어, 비타민 A, C, D, E, B1, B2, B6, B12, 니아신(niacin), 비오틴(biotin), 폴레이트(folate), 판토텐산(panthotenic acid) 등을 포함할 수 있다. 또한, 아연(Zn), 철(Fe), 칼슘(Ca), 크롬(Cr), 마그네슘(Mg), 망간(Mn), 구리(Cu), 크륨(Cr) 등의 미네랄을 포함할 수 있다. 또한, 라이신, 트립토판, 시스테인, 발린 등의 아미노산을 포함할 수 있다. 또한, 방부제(소르빈산 칼륨, 벤조산나트륨, 살리실산, 데히드로초산나트륨 등), 살균제(표백분과 고도 표백분, 차아염소산나트륨 등), 산화방지제(부틸히드록시아니졸(BHA), 부틸히드록시톨류엔(BHT) 등), 착색제(타르색소 등), 발색제(아질 산 나트륨, 아초산 나트륨 등), 표백제(아황산나트륨), 조미료(MSG 글루타민산나트륨 등), 감미료(둘신, 사이클레메이트, 사카린, 나트륨 등), 향료(바닐린, 락톤류 등), 팽창제(명반, D-주석산수소칼륨 등), 강화제, 유화제, 증점제(호료), 피막제, 검기초제, 거품억제제, 용제, 개량제 등의 식품 첨가물(food additives)을 첨가할 수 있다. 상기 첨가물은 식품의 종류에 따라 선별되고 적절한 양으로 사용된다.The composition comprising the mixed extract of dolea and black radish or a fraction thereof is contained in an amount of 0.01 to 100% by weight, more preferably 1 to 80% by weight, based on the total weight of the food composition. When the food is a beverage, it may be included in a ratio of 1 to 30 g, preferably 3 to 20 g, based on 100 ml. In addition, the composition may include additional ingredients that are commonly used in food compositions to improve odor, taste, vision, and the like. For example, vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, and the like may be included. In addition, it may contain minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu), and chromium (Cr). In addition, amino acids such as lysine, tryptophan, cysteine, and valine may be included. In addition, preservatives (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetate, etc.), disinfectants (bleached and highly bleached, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisole (BHA), butylhydroxytoleuene ( BHT), etc.), colorants (tar color, etc.), coloring agents (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasoning (MSG sodium glutamate, etc.), sweeteners (dulsin, cyclamate, saccharin, sodium, etc.) ), flavorings (vanillin, lactones, etc.), expanding agents (alum, D-potassium hydrogen stannate, etc.), reinforcing agents, emulsifiers, thickeners (thickening agents), coating agents, gum base agents, foam inhibitors, solvents, improving agents, etc. additives) can be added. The additive is selected according to the type of food and used in an appropriate amount.
한편, 상기 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는 식품 조성물을 이용하여 염증성 질환 또는 비만의 예방 또는 개선용 기능성식품을 제조할 수 있다.On the other hand, it is possible to prepare a functional food for preventing or improving inflammatory diseases or obesity by using a food composition comprising the mixed extract of Dole and black radish or a fraction thereof.
구체적인 예로, 상기 조성물을 이용하여 농산물, 축산물 또는 수산물의 특성을 살려 변형시키는 동시에 저장성을 좋게한 가공식품을 제조할 수 있다. 이런 가공식품에는 예들 들어, 과자, 음료, 주류, 발효식품, 통조림, 우유가공식품, 육륙가공식품, 국수 등을 포함한다. 과자는 비스킷, 파이, 케익, 빵, 캔디, 젤리, 껌, 시리얼(곡물푸레이크 등의 식사대용품류 포함) 등을 포함한다. 음료는 탄산음료, 기능성이온음료, 쥬스(예들 들어, 사과, 배, 포도, 알로에, 감귤, 복숭아, 당근, 토마토 쥬스 등), 식혜 등을 포함한다. 주류는 청주, 위스키, 소주, 맥주, 양주, 과실주 등을 포함한다. 발효식품은 간장, 된장, 고추장 등을 포함한다. 통조림은 수산물 통조림(예들 들어, 참치, 고등어, 꽁치, 소라 통조림 등), 축산물 통조림(쇠고기, 돼지고기, 닭고기, 칠면조 통조림 등), 농산물 통조림(옥수수, 복숭아, 파일애플 통조림 등)을 포함한다. 우유가공식품은 치즈, 버터, 요구르트 등을 포함한다. 육류가공식품은 돈까스, 비프까스, 치킨까스, 소세지. 탕수육, 너겟류, 너비아니 등을 포함한다. 밀봉포장생면 등의 국수를 포함한다. 이 외에도 상기 조성물은 레토르트식품, 스프류 등에 사용될 수 있다.As a specific example, it is possible to manufacture processed foods with good storage properties while transforming by utilizing the characteristics of agricultural products, livestock products, or aquatic products by using the composition. Such processed foods include, for example, sweets, beverages, alcoholic beverages, fermented foods, canned foods, processed milk foods, processed meat products, noodles, and the like. Confectionery includes biscuits, pies, cakes, bread, candy, jelly, gum, and cereals (including meal substitutes such as grain flour). Beverages include carbonated beverages, functional ionic beverages, juices (for example, apples, pears, grapes, aloe, tangerines, peaches, carrots, tomato juice, etc.), and sikhye. Alcoholic beverages include sake, whiskey, shochu, beer, Western liquor, and fruit liquor. Fermented foods include soy sauce, soybean paste, and red pepper paste. Canned food includes canned seafood (e.g., canned tuna, mackerel, saury, conch, etc.), canned livestock products (beef, pork, chicken, canned turkey, etc.), canned agricultural products (corn, peaches, canned pile apples, etc.). Processed milk foods include cheese, butter, and yogurt. Meat processed foods are pork cutlet, beef cutlet, chicken cutlet, and sausage. Includes sweet and sour pork, nuggets, and width. Includes noodles such as sealed and packaged fresh noodles. In addition to this, the composition may be used for retort foods, soups, and the like.
본 발명의 용어 "기능성식품(functional food)"이란, 특정보건용 식품(food for special health use, FoSHU)와 동일한 용어로, 영양 공급 외에도 생체조절기능이 효율적으로 나타나도록 가공된 의학, 의료효과가 높은 식품을 의미한다. 본 발명에서 용어,“건강식품(health food)"은 일반식품에 비해 적극적인 건강유지나 증진 효과를 가지는 식품을 의미하고, 건강보조식품(health supplement food)는 건강 보조 목적의 식품을 의미한다. 경우에 따라, 기능성식품, 건강식품, 건강보조식품의 용어는 호용된다. 상기 식품은 염증성 질환 또는 비만의 예방 또는 개선에 유용한 효과를 얻기 위하여 정제, 캅셀, 분말, 과립, 액상, 환 등의 다양한 형태로 제조될 수 있다.The term "functional food" of the present invention is the same term as food for special health use (FoSHU). In addition to supplying nutrition, the term "functional food" is processed to effectively display biomodulatory functions. Means high food. In the present invention, the term "health food" refers to a food having an active health maintenance or promotion effect compared to general food, and a health supplement food refers to a food for the purpose of health supplementation. Accordingly, the terms of functional food, health food, health supplement food are used favorably.The food is in various forms such as tablets, capsules, powders, granules, liquids, pills, etc. to obtain useful effects in the prevention or improvement of inflammatory diseases or obesity. Can be manufactured.
본 발명의 또 다른 실시양태는 돌외 및 검정무의 혼합추출물 또는 그의 분획물을 포함하는, 염증성 질환 또는 비만 개선용 사료 조성물을 제공한다.Another embodiment of the present invention provides a feed composition for improving inflammatory diseases or obesity, comprising a mixed extract of dole and black radish or a fraction thereof.
상기 사료 조성물은 사료 첨가제를 포함할 수 있다. 본 발명의 사료첨가제는 사료관리법상의 보조사료에 해당한다.The feed composition may include a feed additive. The feed additive of the present invention corresponds to an auxiliary feed in the feed management law.
본 발명에서 용어, "사료"는 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분을 의미할 수 있다.In the present invention, the term "feed" may mean any natural or artificial diet, one meal meal, etc., or a component of the one meal meal, for animals to eat, ingest, and digest.
상기 사료의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 사료를 사용할 수 있다. 상기 사료의 비제한적인 예로는, 곡물류, 근과류, 식품 가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 또는 곡물 부산물류 등과 같은 식물성 사료; 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질류, 동물성 플랑크톤류 또는 음식물 등과 같은 동물성 사료를 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상을 혼합하여 사용될 수 있다.The kind of feed is not particularly limited, and feed commonly used in the art may be used. Non-limiting examples of the feed include vegetable feeds such as grains, root fruits, food processing by-products, algae, fiber, pharmaceutical by-products, oils and fats, starches, meals or grain by-products; And animal feeds such as proteins, inorganic logistics, oils, minerals, oils and fats, single cell proteins, zooplanktons or food. These may be used alone or in combination of two or more.
본 발명에서 제공하는 돌외 및 검정무의 혼합추출물은 돌외 추출물이 나타내는 독성을 완화시킬 수 있으면서도, 우수한 항염증 또는 항비만 효과를 나타내므로, 다양한 치료제 및 건강식품의 개발에 널리 활용될 수 있을 것이다. The mixed extract of dole and black radish provided by the present invention can alleviate the toxicity of the dole extract and exhibit excellent anti-inflammatory or anti-obesity effects, so it will be widely used in the development of various therapeutic agents and health foods.
도 1은 LPS 처리에 의하여 유도된 세포독성에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 그래프이다.
도 2는 LPS 처리에 의하여 유도된 NO 생성에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 그래프이다.
도 3a는 LPS 처리에 의하여 발현된 COX-2의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.
도 3b는 LPS 처리에 의하여 발현된 iNOS의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.
도 3c는 LPS 처리에 의하여 발현된 인산화된 p65(p-p65)의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.
도 3d는 LPS 처리에 의하여 발현된 JNK의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.
도 4는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)이 3T3-L1 세포에 독성을 나타내는지의 여부를 확인한 결과를 나타내는 그래프이다.
도 5a는 3T3-L1 세포의 지방분화에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 분석한 결과를 나타내는 현미경사진이다.
도 5b는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)이 처리된 지방세포에 포함된 중성지방의 수준을 비교한 결과를 나타내는 그래프이다.
도 6a는 3T3-L1 세포로부터 분화된 지방세포에서 발현된 PPARγ( PPARγ1 및 PPARγ2)의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.
도 6b는 3T3-L1 세포로부터 분화된 지방세포에서 발현된 aP2의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.
도 7은 검정무 추출물(BRE), 돌외 추출물(GYE), 혼합추출물1(MIX1), 및 혼합추출물2(MIX2)이 지방세포에 독성을 나타내는지의 여부를 확인한 결과를 나타내는 그래프이다.
도 8은 검정무 추출물(BRE), 돌외 추출물(GYE), 혼합추출물1(MIX1), 및 혼합추출물2(MIX2)이 나타내는 지방생성 억제능을 평가한 결과를 나타내는 도표로서, (-)는 지방생성 억제능이 없음을 나타내고; (+)는 30% 이상의 지방생성 억제능을 나타내며; (++)는 70% 이상의 지방생성 억제능을 나타내고; (+++)는 90% 이상의 지방생성 억제능을 나타낸다.1 is a graph showing the results of comparing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on the cytotoxicity induced by LPS treatment.
2 is a graph showing the results of comparing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on NO production induced by LPS treatment.
Figure 3a is a Western blot analysis result showing the result of comparing the effect of black radish extract (BRE), Dole extract (GYE), and mixed extract (MIX) on the level of COX-2 expressed by LPS treatment and a quantitative graph thereof to be.
3B is a Western blot analysis result showing the result of comparing the effect of black radish extract (BRE), Dole extract (GYE), and mixed extract (MIX) on the level of iNOS expressed by LPS treatment, and a quantitative graph thereof.
Figure 3c is a Western blot analysis showing the results of comparing the effect of black radish extract (BRE), Dole extract (GYE) and mixed extract (MIX) on the level of phosphorylated p65 (p-p65) expressed by LPS treatment It is a result and its quantitative graph.
3D is a Western blot analysis result showing the result of comparing the effect of black radish extract (BRE), Dole extract (GYE), and mixed extract (MIX) on the level of JNK expressed by LPS treatment, and a quantitative graph thereof.
Figure 4 is a graph showing the results of confirming whether or not black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) exhibit toxicity to 3T3-L1 cells.
5A is a photomicrograph showing the results of analyzing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on fat differentiation of 3T3-L1 cells.
5B is a graph showing the results of comparing the levels of triglycerides contained in adipocytes treated with black radish extract (BRE), dole extract (GYE), and mixed extract (MIX).
Figure 6a is a result of comparing the effect of black radish extract (BRE), dole extract (GYE) and mixed extract (MIX) on the level of PPARγ (PPARγ1 and PPARγ2) expressed in adipocytes differentiated from 3T3-L1 cells. This is a Western blot analysis result and a quantitative graph thereof.
6B is a Western blot analysis result showing the results of comparing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on the level of aP2 expressed in adipocytes differentiated from 3T3-L1 cells. And a quantitative graph thereof.
7 is a graph showing the results of confirming whether or not black radish extract (BRE), dole extract (GYE), mixed extract 1 (MIX1), and mixed extract 2 (MIX2) exhibit toxicity to adipocytes.
8 is a chart showing the results of evaluating the ability to inhibit adipogenesis exhibited by black radish extract (BRE), dole extract (GYE), mixed extract 1 (MIX1), and mixed extract 2 (MIX2), where (-) is adipogenesis Indicates no inhibitory ability; (+) represents 30% or more of the ability to inhibit adipogenesis; (++) represents an ability to inhibit adipogenesis of 70% or more; (+++) represents 90% or more of the ability to inhibit adipogenesis.
이하 본 발명을 실시예를 통하여 보다 상세하게 설명한다. 그러나 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것으로 본 발명의 범위가 이들 실시예에 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail through examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
실시예 1: 제주 검정무-돌외 혼합 추출물 제조Example 1: Preparation of mixed extract of Jeju black radish-dol, etc.
제주특별자치도 서귀포시 성산읍 지역에서 2016년 2월에 수확한 제주산 검정무와 돌외를 각각 수돗물로 3회 이상 세척하여 70 ℃ ~ 80 ℃ 열수에 10분간 침지한 후, 열풍건조를 시킨 후 분쇄하였다. 분쇄된 시료 200 g을 70% 에탄올 3 L에 24시간 동안 추출교반하면서 침출시키며, 다시 초음파 분쇄를 거친 후 여과지로 여과하여 40℃, 90 rpm 속도에서 감압회전농축기로 농축시킨 후 이를 동결 건조시켜 분말상의 검정무 추출물(BRE) 및 돌외 추출물(GYE)을 각각 제작하였다. 끝으로, 상기 수득한 검정무 추출물과 돌외 추출물을 9:1(w/w)로 혼합하고, 이를 20% DMSO에 녹인 후 0.2μM syringe filter를 사용하여 여과한 최종산물을 혼합추출물(MIX) 시료로서 사용하였다.Jeju black radish and dolphin harvested in Seongsan-eup, Seogwipo-si, Jeju Special Self-Governing Province in February 2016 were washed three or more times with tap water, immersed in hot water at 70°C to 80°C for 10 minutes, dried with hot air, and pulverized. 200 g of the pulverized sample is leached with extraction and stirring for 24 hours in 3 L of 70% ethanol. After ultrasonic pulverization, it is filtered through a filter paper, concentrated with a vacuum rotary concentrator at 40°C and 90 rpm, and freeze-dried to form a powder. The black radish extract (BRE) and dole extract (GYE) were prepared, respectively. Finally, the obtained black radish extract and Dole extract were mixed at 9:1 (w/w), dissolved in 20% DMSO, and then filtered using a 0.2 μM syringe filter, and the final product was mixed extract (MIX) sample. Was used as.
실시예 2: RAW264.7 세포를 이용한 혼합추출물의 효과 검증Example 2: Verification of the effect of mixed extracts using RAW264.7 cells
실시예 2-1: 세포독성에 미치는 혼합추출물의 효과Example 2-1: Effect of mixed extract on cytotoxicity
마우스 대식세포인 RAW264.7 세포를 DMEM 배지(1% Penicillin &Streptomycin 100 U/mL, 10% Fetal Bovine Serum, 10% Bovine Serum)에서 37℃ 및 5% CO^2 조건에서 배양하고, 3 내지 4일 간격으로 계대배양하였다.Mouse macrophage RAW264.7 cells were cultured in DMEM medium (1% Penicillin & Streptomycin 100 U/mL, 10% Fetal Bovine Serum, 10% Bovine Serum) at 37° C. and 5% CO^2, and 3-4 days It was subcultured at intervals.
세포독성실험은 MTT assay를 사용하였다. 대략적으로, RAW264.7 세포를 48 well plate(5×10⁴cells/ml)에서 24시간 동안 배양한 다음, LPS (0.5μg/mL)와 각 시료(BRE, GYE 및 MIX)를 농도별로 처리하고(25 내지 1,000 ㎍/㎖) 다시 24 및 48시간 동안 배양하였다. 이어, 상기 배양물에 MTT(3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide)(400 μg/mL)시약을 가하고 37℃에 4시간 배양한 후, 배지를 제거하고 DMSO로 formazan을 녹여 shaker에 1시간 반응시켰다. microplate reader(Bio-TEK Instruments Inc., Vermont, WI, USA)를 사용하여 540 nm에서 흡광도를 측정하였다(도 1). The cytotoxicity test was performed using the MTT assay. Roughly, RAW264.7 cells were incubated for 24 hours in 48 well plates (5×10⁴cells/ml), then LPS (0.5 μg/mL) and each sample (BRE, GYE and MIX) were treated by concentration (25 To 1,000 μg/ml) and cultured again for 24 and 48 hours. Then, MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (400 μg/mL) reagent was added to the culture and incubated at 37° C. for 4 hours. , The medium was removed, formazan was dissolved in DMSO, and reacted with a shaker for 1 hour. Absorbance was measured at 540 nm using a microplate reader (Bio-TEK Instruments Inc., Vermont, WI, USA) (FIG. 1).
도 1은 LPS 처리에 의하여 유도된 세포독성에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 그래프이다.1 is a graph showing the results of comparing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on the cytotoxicity induced by LPS treatment.
도 1에서 보듯이, LPS에 의해 유도된 세포독성은 검정무 추출물에 의해 해소되었으나, 돌외 추출물 및 혼합추출물은 LPS에 의한 세포독성에 별다른 영향을 미치지 못함을 확인하였다.As shown in FIG. 1, the cytotoxicity induced by LPS was resolved by the black radish extract, but it was confirmed that the Dole extract and the mixed extract did not have a significant effect on the cytotoxicity caused by LPS.
실시예 2-2: NO(Nitric oxide) 생성에 미치는 혼합추출물의 효과Example 2-2: Effect of mixed extract on the production of NO (Nitric oxide)
RAW264.7 세포를 48 well plate(1.5×10^5 cells/ml)에 접종하고 18시간 동안 사전배양하였다. 이 후 농도별로 각 추출물 처리하고 1시간 뒤, LPS (0.5μg/mL)를 처리하여 24시간 배양하였다. 배양이 종료된 후, 각 세포에서 생성된 NO의 수준을 측정하였다. 이때, 상기 NO의 수준은 Griess 시약 1 % (w/v) sulfanilamide, 0.1% (w/v) naphylethylenediamine in 2.5% (v/v) phosphoric acid]을 이용하여 세포배양액 중에 존재하는 NO2-의 형태로 측정하였는데, 대략적으로, 세포배양 상등액 100 ㎕와 Griess 시약 100 ㎕를 혼합하여 96 well plates에서 10분 동안 반응시킨 후 540 nm에서 흡광도를 측정하였다(도 2). 생성된 NO의 양은 sodium nitrite (NaNO2)를 standard로 사용하여 비교하였다.RAW264.7 cells were inoculated into 48 well plates (1.5×10^5 cells/ml) and pre-cultured for 18 hours. Thereafter, each extract was treated by concentration, and 1 hour later, LPS (0.5 μg/mL) was treated and cultured for 24 hours. After the culture was completed, the level of NO produced in each cell was measured. At this time, the level of NO is in the form of NO2- present in the cell culture solution using Griess reagent 1% (w/v) sulfanilamide, 0.1% (w/v) naphylethylenediamine in 2.5% (v/v) phosphoric acid]. Approximately, 100 µl of the cell culture supernatant and 100 µl of Griess reagent were mixed and reacted in 96 well plates for 10 minutes, and the absorbance was measured at 540 nm (FIG. 2). The amount of NO produced was compared using sodium nitrite (NaNO2) as a standard.
도 2는 LPS 처리에 의하여 유도된 NO 생성에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 그래프이다.2 is a graph showing the results of comparing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on NO production induced by LPS treatment.
도 2에서 보듯이, LPS를 단독으로 처리하면, LPS 미처리 세포에 비하여 78배 증가된 수준으로 NO가 생성되었으나, 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 NO의 수준이 감소됨을 확인하였다. 특히, 검정무 추출물(BRE) 및 돌외 추출물(GYE)을 처리한 경우 보다도 혼합추출물(MIX)을 처리한 경우에 NO 수준이 크게 감소됨을 확인하였다. 이때, 상기 혼합추출물(MIX)은 검정무 추출물(BRE) 보다 2배 정도 NO 합성을 억제하는 효과를 나타내었고, 돌외 추출물(GYE) 보다는 약 10% 향상된 NO 합성을 억제하는 효과를 나타냄을 알 수 있었다.As shown in FIG. 2, when LPS was treated alone, NO was produced at a level that was 78 times higher than that of LPS untreated cells, but when treated with black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) , It was confirmed that the level of NO increased by LPS was reduced. In particular, it was confirmed that the NO level was significantly reduced when the mixed extract (MIX) was treated than when the black radish extract (BRE) and dole extract (GYE) were treated. At this time, it can be seen that the mixed extract (MIX) exhibited an effect of inhibiting NO synthesis about twice as much as that of black radish extract (BRE), and exhibited an effect of inhibiting NO synthesis, which was improved by about 10% compared to the dole extract (GYE). there was.
실시예 2-3: 염증관련 인자의 발현에 미치는 혼합추출물의 효과Example 2-3: Effect of mixed extract on expression of inflammation-related factors
RAW264.7 세포를 48 well plate(1.5×10^5 cells/ml)에 접종하고 18시간 동안 사전배양하였다. 이 후 농도별로 각 추출물 처리하고 1시간 뒤, LPS (0.5μg/mL)를 처리하여 24시간 배양하였다. 배양이 종료된 후, 각 세포에서 발현된 염증관련 인자(COX-2, iNOS, p-p65 및 JNK)의 수준을 웨스턴블럿 분석을 통해 측정하였다(도 3a 내지 3d).RAW264.7 cells were inoculated into 48 well plates (1.5×10^5 cells/ml) and pre-cultured for 18 hours. Thereafter, each extract was treated by concentration, and 1 hour later, LPS (0.5 μg/mL) was treated and cultured for 24 hours. After the culture was completed, the levels of inflammation-related factors (COX-2, iNOS, p-p65, and JNK) expressed in each cell were measured through Western blot analysis (FIGS. 3A to 3D ).
도 3a는 LPS 처리에 의하여 발현된 COX-2의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.3A is a Western blot analysis result showing the result of comparing the effect of black radish extract (BRE), Dole extract (GYE), and mixed extract (MIX) on the level of COX-2 expressed by LPS treatment, and a quantitative graph thereof to be.
도 3a에서 보듯이, LPS를 단독으로 처리하면, LPS 미처리 세포에 비하여 현저히 증가된 수준으로 Cox-2의 발현수준이 증가되었으나, 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 Cox-2의 발현수준이 감소됨을 확인하였다. 특히, 검정무 추출물(BRE) 및 혼합추출물(MIX)을 처리한 경우 보다도 돌외 추출물(GYE)을 처리한 경우에 Cox-2의 발현수준이 상대적으로 더욱 감소됨을 확인하였다.As shown in Figure 3a, treatment with LPS alone increased the expression level of Cox-2 to a significantly increased level compared to cells not treated with LPS, but black radish extract (BRE), dole extract (GYE), and mixed extract (MIX ), it was confirmed that the expression level of Cox-2 increased by LPS decreased. In particular, it was confirmed that the expression level of Cox-2 was relatively further reduced when treated with GYE extract than when treated with black radish extract (BRE) and mixed extract (MIX).
도 3b는 LPS 처리에 의하여 발현된 iNOS의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.Figure 3b is a Western blot analysis result showing the result of comparing the effect of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on the level of iNOS expressed by LPS treatment, and a quantitative graph thereof.
도 3b에서 보듯이, LPS를 단독으로 처리하면, LPS 미처리 세포에 비하여 현저히 증가된 수준으로 iNOS의 발현수준이 증가되었으나, 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 Cox-2의 발현수준이 현저하게 감소됨을 확인하였다. 상기 돌외 추출물(GYE)을 처리한 경우 보다도 혼합추출물(MIX)을 처리한 경우에 iNOS의 발현수준이 상대적으로 더욱 감소됨을 확인하였다.As shown in Fig. 3b, when LPS was treated alone, the expression level of iNOS was increased to a significantly increased level compared to LPS untreated cells, but when treated with a dole extract (GYE) and a mixed extract (MIX), it was increased by LPS. It was confirmed that the expression level of Cox-2 was significantly reduced. It was confirmed that the expression level of iNOS was relatively further reduced when the mixed extract (MIX) was treated than when the dole extract (GYE) was treated.
도 3c는 LPS 처리에 의하여 발현된 인산화된 p65(p-p65)의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.Figure 3c is a Western blot analysis showing the results of comparing the effect of black radish extract (BRE), Dole extract (GYE) and mixed extract (MIX) on the level of phosphorylated p65 (p-p65) expressed by LPS treatment It is a result and its quantitative graph.
도 3c에서 보듯이, LPS를 단독으로 처리하면, LPS 미처리 세포에 비하여 현저히 증가된 수준으로 p65의 인산화수준이 증가되었으나, 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 p65의 인산화수준이 감소됨을 확인하였다. 상기 검정무 추출물(BRE)은 LPS에 의해 증가된 p65의 인산화수준을 일부 감소시켰으나, 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 p65의 인산화수준이 현저하게 감소됨을 확인하였다. 상기 혼합추출물(MIX)을 처리한 경우 보다도 돌외 추출물(GYE)을 처리한 경우에 p65의 인산화수준이 상대적으로 더욱 감소됨을 확인하였다.As shown in Fig. 3c, when LPS was treated alone, the phosphorylation level of p65 was increased to a significantly increased level compared to cells not treated with LPS, but black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) were used. Upon treatment, it was confirmed that the phosphorylation level of p65, which was increased by LPS, decreased. The black radish extract (BRE) partially reduced the phosphorylation level of p65, which was increased by LPS, but when treated with the Dole extract (GYE) and mixed extract (MIX), the phosphorylation level of p65 increased by LPS was significantly reduced. Was confirmed. It was confirmed that the phosphorylation level of p65 was relatively further reduced when GYE was treated than when the mixed extract (MIX) was treated.
도 3d는 LPS 처리에 의하여 발현된 JNK의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.3D is a Western blot analysis result showing the result of comparing the effect of black radish extract (BRE), Dole extract (GYE), and mixed extract (MIX) on the level of JNK expressed by LPS treatment, and a quantitative graph thereof.
도 3d에서 보듯이, LPS를 단독으로 처리하면, LPS 미처리 세포에 비하여 현저히 증가된 수준으로 JNK의 발현수준이 증가되었으나, 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 JNK의 발현수준이 감소됨을 확인하였다. 상기 검정무 추출물(BRE)은 LPS에 의해 증가된 JNK의 발현수준을 일부 감소시켰으나, 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, LPS에 의해 증가된 JNK의 발현수준이 현저하게 감소됨을 확인하였다. 상기 돌외 추출물(GYE)을 처리한 경우 보다도 혼합추출물(MIX)을 처리한 경우에 JNK의 발현수준이 상대적으로 더욱 감소됨을 확인하였다.As shown in FIG. 3D, when LPS was treated alone, the expression level of JNK was increased to a significantly increased level compared to LPS-untreated cells, but black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) were used. When treated, it was confirmed that the expression level of JNK increased by LPS was reduced. The black radish extract (BRE) partially reduced the expression level of JNK increased by LPS, but when treated with a dole extract (GYE) and mixed extract (MIX), the expression level of JNK increased by LPS was significantly reduced. Was confirmed. It was confirmed that the expression level of JNK was relatively further reduced when the mixed extract (MIX) was treated than when the dole extract (GYE) was treated.
상기 도 3a 내지 3d의 결과로부터, 혼합추출물은 검정무 추출물 및 돌외 추출물에 비하여, NO생성 억제, Cox-2 발현 억제, iNOS 발현 억제, P65 인산화 억제를 통한 NF-kB 신호전달 차단 효과, JNK 인산화 억제효과가 현저한 차이를 나타냄을 확인하였으므로, 상기 혼합추출물을 염증억제 조성물로 사용할 수 있음을 알 수 있었다. From the results of FIGS. 3A to 3D, the mixed extract inhibits NO production, inhibits Cox-2 expression, inhibits iNOS expression, inhibits NF-kB signaling through inhibition of P65 phosphorylation, compared to the black radish extract and the Dole extract, and JNK phosphorylation. Since it was confirmed that the inhibitory effect showed a remarkable difference, it was found that the mixed extract can be used as an anti-inflammatory composition.
실시예 3: 3T3-L1 세포를 이용한 혼합추출물의 효과 검증Example 3: Verification of the effect of mixed extracts using 3T3-L1 cells
실시예 3-1: 세포독성에 미치는 혼합추출물의 효과Example 3-1: Effect of mixed extract on cytotoxicity
마우스 전지방세포 3T3-L1 세포를 DMEM 배지(1% Penicillin &Streptomycin 100 U/mL, 10% Fetal Bovine Serum, 10% Bovine Serum)에서 37℃ 및 5% CO^2 조건에서 배양하고, 3 내지 4일 간격으로 계대배양하였다.Mouse protozoan 3T3-L1 cells were cultured in DMEM medium (1% Penicillin & Streptomycin 100 U/mL, 10% Fetal Bovine Serum, 10% Bovine Serum) at 37° C. and 5% CO^2, and 3 to 4 days It was subcultured at intervals.
세포독성실험은 MTT assay를 사용하였다. 대략적으로, 3T3-L1 세포를 48 well plate(5×10⁴cells/ml)에서 24시간 동안 배양한 다음, 각 시료(BRE, GYE 및 MIX)를 농도별로 처리하고(25 내지 500 ㎍/㎖) 다시 24 및 48시간 동안 배양하였다. 이어, 상기 배양물에 MTT(3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide)(400 μg/mL)시약을 가하고 37℃에 4시간 배양한 후, 배지를 제거하고 DMSO로 formazan을 녹여 shaker에 1시간 반응시켰다. microplate reader(Bio-TEK Instruments Inc., Vermont, WI, USA)를 사용하여 540 nm에서 흡광도를 측정하였다(도 4). The cytotoxicity test was performed using the MTT assay. Approximately, 3T3-L1 cells were incubated in 48 well plates (5×10⁴cells/ml) for 24 hours, and then each sample (BRE, GYE and MIX) was treated by concentration (25 to 500 μg/ml) and again 24 And incubated for 48 hours. Then, MTT (3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) (400 μg/mL) reagent was added to the culture and incubated at 37° C. for 4 hours. , The medium was removed, formazan was dissolved in DMSO, and reacted with a shaker for 1 hour. Absorbance was measured at 540 nm using a microplate reader (Bio-TEK Instruments Inc., Vermont, WI, USA) (FIG. 4).
도 4는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)이 3T3-L1 세포에 독성을 나타내는지의 여부를 확인한 결과를 나타내는 그래프이다.Figure 4 is a graph showing the results of confirming whether or not black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) exhibit toxicity to 3T3-L1 cells.
도 4에서 보듯이, 검정무 추출물은 처리농도에 상관없이 24시간 및 48시간에서 세포독성을 나타내지 않았으나, 돌외 추출물은 처리농도에 비례하여 24시간 및 48시간에서 세포독성을 나타내었으며, 혼합추출물은 24시간의 경우에는 처리농도에 비례하여 세포독성을 나타내었으나, 48시간의 경우에는 처리농도에 상관없이 세포독성을 나타내지 않음을 확인하였다.As shown in Figure 4, the black radish extract did not show cytotoxicity at 24 hours and 48 hours regardless of the treatment concentration, but the dole extract showed cytotoxicity at 24 hours and 48 hours in proportion to the treatment concentration, and the mixed extract In the case of 24 hours, it was confirmed that cytotoxicity was shown in proportion to the treatment concentration, but in the case of 48 hours, it was confirmed that it did not show cytotoxicity regardless of the treatment concentration.
따라서, 혼합추출물은 돌외 추출물이 나타내는 세포독성을 완화시키는 효과를 나타냄을 알 수 있었다.Therefore, it was found that the mixed extract exhibited the effect of mitigating the cytotoxicity of the Dole extract.
실시예 3-2: 지방분화에 미치는 혼합추출물의 효과Example 3-2: Effect of mixed extract on fat differentiation
3T3-L1 세포를 24 well(1×10^5 cells/ml)에 분주하고, 100% 포화도(confluency)를 나타내는 시점에 도달한 후, 2일 동안 추가로 배양하였다. 배양이 종료된 상기 세포에 농도별(25 내지 500 ㎍/㎖) 각 시료(BRE, GYE 및 MIX)가 첨가되고, 지방분화유도물질(MDI: 0.5mM 3-Isobutyl-1-methylxanthine(IBMX), 1μM Dexamethasone, 10μg/mlSt.Louis, MO, USA)과 10% FBS가 포함된 DMEM 배지를 가하고 2일 동안 배양한 다음, 10 μg/mL 인슐린과 10% FBS가 포함된 DMEM 배지로 교환하여 2일간 배양하였으며, 그 후 2일 마다 10% FBS가 포함된 DMEM 배지로 교환하여 4일 동안 배양함으로써, 지방세포로의 분화를 유도하였다(도 5a). 이때, 음성대조군은 시료 및 지방분화유도물질을 처리하지 않고 배양한 세포를 사용하고, 양성대조군으로는 시료 없이 지방분화유도물질만을 처리하여 배양한 세포를 사용하였다.3T3-L1 cells were dispensed into 24 wells (1×10^5 cells/ml), and after reaching a time point showing 100% saturation, the cells were further cultured for 2 days. Each sample (BRE, GYE and MIX) by concentration (25 to 500 μg/ml) was added to the cells after the culture was completed, and a fat differentiation inducer (MDI: 0.5mM 3-Isobutyl-1-methylxanthine (IBMX), 1 μM Dexamethasone, 10 μg/ml St.Louis, MO, USA) and DMEM medium containing 10% FBS were added and incubated for 2 days, and then replaced with DMEM medium containing 10 μg/mL insulin and 10% FBS for 2 days. Culture was performed, and then every 2 days, the DMEM medium containing 10% FBS was exchanged and cultured for 4 days to induce differentiation into adipocytes (FIG. 5A). At this time, as a negative control, cells cultured without treatment with a sample and a lipodifferentiation inducer were used, and as a positive control, cells cultured by treating only a fat differentiation inducer without a sample were used.
도 5a는 3T3-L1 세포의 지방분화에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 분석한 결과를 나타내는 현미경사진이다.5A is a photomicrograph showing the results of analyzing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on fat differentiation of 3T3-L1 cells.
지방분화유도가 종료된 후, 분화된 지방세포에 포함된 중성지방의 수준을 측정하기 위하여, Oil Red O(ORO) 염색법을 수행하였다(도 5b).After the induction of fat differentiation was completed, in order to measure the level of triglyceride contained in the differentiated adipocytes, Oil Red O (ORO) staining was performed (FIG. 5B).
대략적으로, 분화된 3T3-L1 세포를 Phosphate Buffered Saline(LPS solution, Daejeon, Korea)로 세척하여 10% formalin(approx. 4% formaldehyde)(Sigma-Aldrich co.)로 30분 이상 고정시켰다. 그 후 증류수로 3회 세척하고 Isopropyl alcohol(Merck, Kenilworth, USA)에 녹인 0.6% Oil Red O(Sigma-Aldrich co.) 염색약으로 30분 동안 염색시킨 후 증류수로 세척하였고 분화된 양상을 현미경을 이용하여 사진 촬영하였다. Isopropyl alcohol에 녹여 520nm에서 흡광도를 측정하였다. Roughly, differentiated 3T3-L1 cells were washed with Phosphate Buffered Saline (LPS solution, Daejeon, Korea) and fixed with 10% formalin (approx. 4% formaldehyde) (Sigma-Aldrich co.) for at least 30 minutes. Then, washed three times with distilled water and stained with 0.6% Oil Red O (Sigma-Aldrich co.) dye dissolved in Isopropyl alcohol (Merck, Kenilworth, USA) for 30 minutes and washed with distilled water. I took a picture. The absorbance was measured at 520 nm by dissolving in isopropyl alcohol.
도 5b는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)이 처리된 지방세포에 포함된 중성지방의 수준을 비교한 결과를 나타내는 그래프이다.5B is a graph showing the results of comparing the levels of triglycerides contained in adipocytes treated with black radish extract (BRE), dole extract (GYE), and mixed extract (MIX).
도 5b에서 보듯이, 지방세포로 분화된 후에는 세포내 중성지방의 수준이 급격히 증가되었으나, 상기 시료를 처리한 경우에는, 세포내 중성지방의 수준이 시료의 농도에 반비례함을 확인하였다. 다만, 상기 중성지방의 수준은 시료별로 차이를 나타내었는데, 혼합추출물을 처리한 경우에 세포내 중성지방의 수준이 상대적으로 가장 낮은 수준을 나타내었고, 검정무 추출물을 처리한 경우에 세포내 중성지방의 수준이 상대적으로 가장 높은 수준을 나타내었는데, 상기 혼합추출물은 검정무 추출물의 4배 및 돌외 추출물의 1.6배의 중성지방 축적억제 효과를 나타내는 것으로 분석되었다.As shown in FIG. 5B, after differentiation into adipocytes, the level of intracellular triglycerides rapidly increased, but when the sample was treated, it was confirmed that the level of intracellular triglycerides was inversely proportional to the concentration of the sample. However, the level of triglyceride was different for each sample, and when the mixed extract was treated, the intracellular triglyceride level was relatively the lowest, and when the black radish extract was treated, the intracellular triglyceride was The level of was the highest level, and the mixed extract was analyzed to have a triglyceride accumulation inhibitory effect of 4 times that of the black radish extract and 1.6 times that of the Dole extract.
따라서, 혼합추출물은 지방합성을 효율적으로 억제하는 효과를 나타냄을 알 수 있었다.Therefore, it was found that the mixed extract exhibited an effect of effectively inhibiting fat synthesis.
실시예 3-3: 지방분화 관련인자의 발현수준에 미치는 혼합추출물의 효과Example 3-3: Effect of mixed extract on the expression level of factors related to fat differentiation
지방분화에 영향을 미치는 유도인자인 PPARγ 및 지방세포의 마커로 알려진 aP2의 발현수준에 미치는 혼합추출물의 효과를 분석하기 위하여, 실시예 3-2의 방법으로 각 시료를 처리한 후, 지방분화를 유도하였다. 지방분화유도가 종료된 후, 각 세포에서 발현된 지방분화 관련인자(PPARγ 및 aP2)의 수준을 웨스턴블럿 분석을 통해 측정하였다(도 6a 및 6b).In order to analyze the effect of the mixed extract on the expression level of PPARγ, an inducer that affects fat differentiation, and aP2 known as a marker of adipocytes, each sample was treated by the method of Example 3-2, and then fat differentiation was performed. Induced. After the induction of lipodifferentiation was completed, the levels of lipodifferentiation-related factors (PPARγ and aP2) expressed in each cell were measured through Western blot analysis (FIGS. 6A and 6B ).
도 6a는 3T3-L1 세포로부터 분화된 지방세포에서 발현된 PPARγ( PPARγ1 및 PPARγ2)의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.Figure 6a is a result of comparing the effect of black radish extract (BRE), dole extract (GYE) and mixed extract (MIX) on the level of PPARγ (PPARγ1 and PPARγ2) expressed in adipocytes differentiated from 3T3-L1 cells. This is a Western blot analysis result and a quantitative graph thereof.
도 6a에서 보듯이, 3T3-L1 세포에 비하여 지방세포에서는 현저히 증가된 수준으로 PPARγ( PPARγ1 및 PPARγ2)의 발현수준이 증가되었으나, 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, 지방세포에서 증가된 PPARγ( PPARγ1 및 PPARγ2)의 발현수준이 감소됨을 확인하였다. 특히, 검정무 추출물(BRE) 및 돌외 추출물(GYE)을 처리한 경우 보다도 혼합추출물(MIX)을 처리한 경우에 PPARγ( PPARγ1 및 PPARγ2)의 발현수준이 상대적으로 더욱 감소됨을 확인하였는데, 상기 혼합추출물은 검정무 추출물의 3배 및 돌외 추출물의 1.3배의 효과를 나타내는 것으로 분석되었다.As shown in Figure 6a, the expression level of PPARγ (PPARγ1 and PPARγ2) was significantly increased in adipocytes compared to 3T3-L1 cells, but black radish extract (BRE), stone extract (GYE), and mixed extract (MIX ), it was confirmed that the expression level of increased PPARγ (PPARγ1 and PPARγ2) in adipocytes decreased. In particular, it was confirmed that the expression level of PPARγ (PPARγ1 and PPARγ2) was relatively further reduced when the mixed extract (MIX) was treated than when the black radish extract (BRE) and dole extract (GYE) were treated. Was analyzed to show the effect of 3 times that of the black radish extract and 1.3 times that of the dole extract.
도 6b는 3T3-L1 세포로부터 분화된 지방세포에서 발현된 aP2의 수준에 미치는 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)의 효과를 비교한 결과를 나타내는 웨스턴블럿 분석결과 및 이의 정량그래프이다.6B is a Western blot analysis result showing the results of comparing the effects of black radish extract (BRE), dole extract (GYE), and mixed extract (MIX) on the level of aP2 expressed in adipocytes differentiated from 3T3-L1 cells. And a quantitative graph thereof.
도 6b에서 보듯이, 3T3-L1 세포에 비하여 지방세포에서는 현저히 증가된 수준으로 aP2의 발현수준이 증가되었으나, 검정무 추출물(BRE), 돌외 추출물(GYE) 및 혼합추출물(MIX)을 처리하면, 지방세포에서 증가된 aP2의 발현수준이 감소됨을 확인하였다. 특히, 검정무 추출물(BRE)의 경우에는 aP2의 발현억제 효과가 낮은 수준을 나타냄에 반하여, 돌외 추출물(GYE)과 혼합추출물(MIX)은 aP2의 발현억제 효과가 매우 높은 수준을 나타내며, 돌외 추출물(GYE) 보다는 혼합추출물(MIX)이 상대적으로 높은 aP2의 발현억제 효과를 나타냄을 확인하였다.As shown in Figure 6b, compared to 3T3-L1 cells, the expression level of aP2 was increased to a significantly increased level in adipocytes, but when black radish extract (BRE), dole extract (GYE) and mixed extract (MIX) were treated, It was confirmed that the expression level of increased aP2 in adipocytes decreased. In particular, in the case of black radish extract (BRE), the expression inhibitory effect of aP2 is low, whereas the dole extract (GYE) and mixed extract (MIX) show a very high level of the expression inhibitory effect of aP2. It was confirmed that the mixed extract (MIX) showed a relatively high inhibitory effect on the expression of aP2 than (GYE).
실시예 4: 혼합추출물내 함량비에 따른 효과Example 4: Effect according to the content ratio in the mixed extract
상기 실시예 2 및 3의 결과로부터, 검정무 추출물과 돌외 추출물을 포함하는 혼합추출물은 염증반응을 억제하고, 지방합성을 억제하는 효과를 나타냄을 확인하였는데, 이러한 효과가 검정무 추출물과 돌외 추출물의 혼합비에 따라 변화되는지의 여부를 분석하였다.From the results of Examples 2 and 3, it was confirmed that the mixed extract containing the black radish extract and the dole extract exhibits an effect of inhibiting the inflammatory reaction and inhibiting the fat synthesis. Whether or not it changes according to the mixing ratio was analyzed.
실시예 4-1: 시료의 준비Example 4-1: Preparation of sample
상기 실시예 1에서 제조한 검정무 추출물(BRE) 및 돌외 추출물(GYE)과, 상기 검정무 추출물(BRE)와 돌외 추출물(GYE)을 9:1(w/w)로 혼합한 혼합추출물1(MIX1), 및 상기 검정무 추출물(BRE)와 돌외 추출물(GYE)을 1:1(w/w)로 혼합한 혼합추출물2(MIX2)를 각각 시료로서 사용하였다.
실시예 4-2: 세포독성 평가Example 4-2: Cytotoxicity evaluation
상기 실시예 3-2의 방법으로 제조된 지방세포에 상기 실시예 4-1에서 준비한 검정무 추출물(BRE), 돌외 추출물(GYE), 혼합추출물1(MIX1), 및 혼합추출물2(MIX2)를 다양한 농도로 처리하고, 실시예 3-1의 MTT assay를 사용하여 세포독성을 분석하였다(도 7).To adipocytes prepared by the method of Example 3-2, the black radish extract (BRE), dole extract (GYE), mixed extract 1 (MIX1), and mixed extract 2 (MIX2) prepared in Example 4-1 were added. Treated at various concentrations, and analyzed for cytotoxicity using the MTT assay of Example 3-1 (FIG. 7).
도 7은 검정무 추출물(BRE), 돌외 추출물(GYE), 혼합추출물1(MIX1), 및 혼합추출물2(MIX2)이 지방세포에 독성을 나타내는지의 여부를 확인한 결과를 나타내는 그래프이다.7 is a graph showing the results of confirming whether or not black radish extract (BRE), dole extract (GYE), mixed extract 1 (MIX1), and mixed extract 2 (MIX2) exhibit toxicity to adipocytes.
도 7에서 보듯이, 검정무 추출물(BRE)은 처리농도에 상관없이 세포독성을 나타내지 않은 반면, 50㎍/㎖ 이상의 농도로 처리된 돌외 추출물(GYE)은 상당한 수준의 세포독성을 나타냄을 확인하였다.As shown in FIG. 7, it was confirmed that the black radish extract (BRE) did not show cytotoxicity regardless of the treatment concentration, whereas the dole extract (GYE) treated with a concentration of 50 µg/ml or more showed a significant level of cytotoxicity. .
또한, 동일한 함량의 돌외 추출물(GYE)을 포함하는 혼합추출물1(MIX1) 및 혼합추출물2(MIX2)의 결과를 비교하면, 50㎍/㎖ 이하의 돌외 추출물(GYE)을 포함하는 혼합추출물은 세포독성을 나타내지 않았으나, 100㎍/㎖의 돌외 추출물(GYE)을 포함하는 혼합추출물은 돌외 추출물(GYE)의 세포독성을 그대로 유지함을 확인하였다.In addition, comparing the results of mixed extract 1 (MIX1) and mixed extract 2 (MIX2) containing the same amount of extradolescent extract (GYE), the mixed extract containing less than 50µg/ml extradolphal extract (GYE) Although it did not show toxicity, it was confirmed that the mixed extract containing 100 μg/ml of Dole extract (GYE) maintained the cytotoxicity of Dole extract (GYE) as it is.
상기 결과로부터, 돌외 추출물(GYE)은 세포독성을 나타내며, 50㎍/㎖ 이하의 돌외 추출물(GYE)은 검정무 추출물(BRE)에 의해 세포독성이 해소될 수 있으나, 100 ㎍/㎖의 돌외 추출물(GYE)은 검정무 추출물(BRE)에 의하여도 세포독성이 해소되지 않음을 알 수 있었다.From the above results, extradolescent extract (GYE) exhibits cytotoxicity, and cytotoxicity of 50µg/ml or less of extradolescent extract (GYE) can be eliminated by black radish extract (BRE), but 100µg/ml extradolescent extract (GYE) was found that cytotoxicity was not resolved even by black radish extract (BRE).
따라서, 50㎍/㎖ 이하의 돌외 추출물(GYE)을 포함하는 혼합추출물을 사용함이 바람직할 것으로 분석되었다.Therefore, it was analyzed that it would be preferable to use a mixed extract containing 50 µg/ml or less of Dole extract (GYE).
실시예 4-3: 지방합성 억제효능 평가Example 4-3: Evaluation of liposynthesis inhibitory effect
상기 실시예 4-1에서 준비된 각 시료를 다양한 농도로 처리하는 것을 제외하고는, 실시예 3-2의 방법을 수행하여, 상기 각 시료가 나타내는 지방생성 억제능을 평가하였다(도 8).Except for treating each sample prepared in Example 4-1 at various concentrations, the method of Example 3-2 was performed to evaluate the ability of each sample to inhibit adipogenesis (FIG. 8).
도 8은 검정무 추출물(BRE), 돌외 추출물(GYE), 혼합추출물1(MIX1), 및 혼합추출물2(MIX2)이 나타내는 지방생성 억제능을 평가한 결과를 나타내는 도표로서, (-)는 지방생성 억제능이 없음을 나타내고; (+)는 30% 이상의 지방생성 억제능을 나타내며; (++)는 70% 이상의 지방생성 억제능을 나타내고; (+++)는 90% 이상의 지방생성 억제능을 나타낸다.8 is a chart showing the results of evaluating the ability to inhibit adipogenesis exhibited by black radish extract (BRE), dole extract (GYE), mixed extract 1 (MIX1), and mixed extract 2 (MIX2), where (-) is adipogenesis Indicates no inhibitory ability; (+) represents 30% or more of the ability to inhibit adipogenesis; (++) represents an ability to inhibit adipogenesis of 70% or more; (+++) represents 90% or more of the ability to inhibit adipogenesis.
도 8에서 보듯이, 125 ㎍/㎖의 검정무 추출물(BRE) 및 12.5 ㎍/㎖의 돌외 추출물(GYE)은 각각 지방억제능을 나타내지 않지만, 이들을 혼합한 혼합추출물1(MIX1)은 30% 이상의 지방생성 억제능을 나타냄을 확인하였다. As shown in Fig. 8, 125 ㎍ / ㎖ of black radish extract (BRE) and 12.5 ㎍ / ㎖ of Dole extract (GYE) do not show fat inhibitory activity, respectively, but the mixed extract 1 (MIX1) of these mixtures is more than 30% fat It was confirmed that the production inhibitory ability was shown.
또한, 혼합추출물1(MIX1)은 모든 경우에 지방생성 억제능을 나타내지만, 혼합추출물2(MIX2)는 일정 농도 이상에서만 지방생성 억제능을 나타냄을 확인하였다.In addition, it was confirmed that the mixed extract 1 (MIX1) showed the ability to inhibit adipogenesis in all cases, but the mixed extract 2 (MIX2) showed the ability to inhibit adipogenesis only at a certain concentration or higher.
상술한 결과를 종합하면, 검정무 추출물과 돌외 추출물을 포함하는 혼합추출물은 염증반응을 억제하고, 지방합성을 억제하는 효과를 나타내며, 이러한 효과는 검정무 추출물과 돌외 추출물이 9:1(w/w)로 혼합된 경우에만 나타남을 알 수 있었다.Summarizing the above results, the mixed extract containing the black radish extract and the dole extract exhibits an effect of suppressing the inflammatory reaction and inhibiting the synthesis of fat, and such an effect is that the black radish extract and the Dole extract are 9:1 It can be seen that it appears only when mixed with w).
이상의 설명으로부터, 본 발명이 속하는 기술분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시 예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art to which the present invention pertains will understand that the present invention can be implemented in other specific forms without changing the technical spirit or essential features thereof. In this regard, the embodiments described above are illustrative in all respects and should be understood as not limiting. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the claims to be described later rather than the above detailed description and equivalent concepts are included in the scope of the present invention.
Claims (12)
A pharmaceutical composition for the prevention or treatment of inflammatory diseases or obesity, comprising a mixed extract of dole and black radish or a fraction thereof.
상기 혼합 추출물은 돌외 또는 검정무를 물, 탄소수 1 내지 4의 저급 알코올 및 이들의 혼합용매로 구성된 군으로부터 선택되는 용매로 각각 추출한 후, 이를 혼합하여 수득한 것인, 약학 조성물.
The method of claim 1,
The mixed extract is obtained by extracting each of the dorsal or black radish with a solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof, and then mixing them.
상기 돌외 추출물과 검정무 추출물의 혼합비는 1:5 내지 1:9(w/w)인 것인, 약학 조성물.
The method of claim 2,
The mixing ratio of the dole extract and the black radish extract is 1:5 to 1:9 (w/w), the pharmaceutical composition.
상기 혼합 추출물은 돌외 및 검정무의 혼합물을 물, 탄소수 1 내지 4의 저급 알코올 및 이들의 혼합용매로 구성된 군으로부터 선택되는 용매로 추출하여 수득한 것인, 약학 조성물.
The method of claim 1,
The mixed extract is obtained by extracting a mixture of dole and black radish with a solvent selected from the group consisting of water, a lower alcohol having 1 to 4 carbon atoms, and a mixed solvent thereof.
상기 돌외와 검정무의 혼합비는 1:5 내지 1:9(w/w)인 것인, 약학 조성물.
The method of claim 4,
The mixing ratio of the stone and black radish is 1:5 to 1:9 (w / w) will, the pharmaceutical composition.
상기 분획물은 상기 혼합 추출물을 용매 분획법, 한외여과 분획법, 크로마토그래피 분획법 및 이들의 조합으로 구성된 군으로부터 선택되는 방법에 적용하여 수득한 분획물인 것인, 약학 조성물.
The method of claim 1,
The fraction is a fraction obtained by applying the mixed extract to a method selected from the group consisting of a solvent fractionation method, an ultrafiltration fractionation method, a chromatography fractionation method, and a combination thereof.
상기 혼합 추출물은 NO 생성억제, Cox-2 발현억제, iNOS 발현억제, p65 인산화억제 및 JNK 인산화억제 효과를 나타내는 것인, 약학 조성물.
The method of claim 1,
The mixed extract will exhibit NO production inhibition, Cox-2 expression inhibition, iNOS expression inhibition, p65 phosphorylation inhibition and JNK phosphorylation inhibitory effects.
상기 혼합 추출물은 지방세포 분화억제, 지방세포내 중성지방 축적억제, PPARγ 발현억제 및 aP2 발현억제 효과를 나타내는 것인, 약학 조성물.
The method of claim 1,
The mixed extract will exhibit the effect of inhibiting adipocyte differentiation, inhibiting accumulation of triglycerides in adipocytes, inhibiting PPARγ expression, and inhibiting aP2 expression.
상기 약학 조성물은 약학적으로 허용되는 담체, 부형제 또는 희석제를 추가로 포함하는 것인, 약학 조성물.
The method of claim 1,
The pharmaceutical composition further comprises a pharmaceutically acceptable carrier, excipient, or diluent.
A method for preventing or treating an inflammatory disease or obesity, comprising administering the pharmaceutical composition of any one of claims 1 to 9 to an individual other than humans, which has or is likely to develop an inflammatory disease or obesity.
A food composition for improving inflammatory disease or obesity, comprising a mixed extract of dole and black radish or a fraction thereof.
A feed composition for improving inflammatory disease or obesity, comprising a mixed extract of dole and black radish or a fraction thereof.
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| KR1020190134238A KR20210050085A (en) | 2019-10-28 | 2019-10-28 | Pharmaceutical composition comprising mixed extract of Gynostemma phetaphyllum and black radish |
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| KR (1) | KR20210050085A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100930580B1 (en) | 2009-07-17 | 2009-12-09 | 주식회사 티지 바이오텍 | The method for preparing gynostemma pentaphyllum extract with increasing damulin a and damulin b contents, and a pharmaceutical compositions of the same for treating metabolic disease |
| KR101212346B1 (en) | 2005-06-09 | 2012-12-13 | 애경산업(주) | Cosmetic compositions for preventing and curing acne |
| KR101910156B1 (en) | 2016-10-26 | 2018-10-19 | 재단법인 제주테크노파크 | Anti-inflammatory Composition Using an Extract of Black Radish |
| KR20190084927A (en) | 2019-07-08 | 2019-07-17 | 재단법인 제주테크노파크 | Composition for preventing or treating obesity or fatty liver comprising extract of Raphanus sativus L. var niger as an active ingredient |
-
2019
- 2019-10-28 KR KR1020190134238A patent/KR20210050085A/en not_active Application Discontinuation
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101212346B1 (en) | 2005-06-09 | 2012-12-13 | 애경산업(주) | Cosmetic compositions for preventing and curing acne |
| KR100930580B1 (en) | 2009-07-17 | 2009-12-09 | 주식회사 티지 바이오텍 | The method for preparing gynostemma pentaphyllum extract with increasing damulin a and damulin b contents, and a pharmaceutical compositions of the same for treating metabolic disease |
| KR101910156B1 (en) | 2016-10-26 | 2018-10-19 | 재단법인 제주테크노파크 | Anti-inflammatory Composition Using an Extract of Black Radish |
| KR20190084927A (en) | 2019-07-08 | 2019-07-17 | 재단법인 제주테크노파크 | Composition for preventing or treating obesity or fatty liver comprising extract of Raphanus sativus L. var niger as an active ingredient |
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