KR20080004811A - Extract of angelica archangelica having anti-asthma activity - Google Patents
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- KR20080004811A KR20080004811A KR1020060063589A KR20060063589A KR20080004811A KR 20080004811 A KR20080004811 A KR 20080004811A KR 1020060063589 A KR1020060063589 A KR 1020060063589A KR 20060063589 A KR20060063589 A KR 20060063589A KR 20080004811 A KR20080004811 A KR 20080004811A
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- extract
- asthma
- angelica archangelica
- interleukin
- cells
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Abstract
Description
도 1은 본 발명에 따른 신선초 추출물의 산화스트레스에 의한 세포막 손상 저해 효과를 검증한 사진이다. 1 is a photograph verifying the inhibitory effect of cell membrane damage by oxidative stress of the extract of fresh vinegar according to the present invention.
도 2는 본 발명에 따른 신선초 추출물의 산화스트레스에 의한 세포손상 저해 효과를 검증한 엠티티(MTT) 시험의 결과이다. Figure 2 is the result of the MTT test to verify the cell damage inhibition effect by the oxidative stress of the fresh vinegar extract according to the present invention.
도 3은 본 발명에 따른 신선초 추출물의 마우스 허파에서의 CD4 파지티브 세포의 발현 억제 효과를 나타낸 사진이다. Figure 3 is a photograph showing the effect of inhibiting the expression of CD4 positive cells in mouse lungs of the extract of Sinchocho according to the present invention.
도 4는 본 발명에 따른 신선초 추출물의 마우스 허파에서의 인터루킨-4 사이토카인 분비 억제 효과를 나타낸 사진이다. Figure 4 is a photograph showing the inhibitory effect of interleukin-4 cytokine secretion in mouse lungs of Sinchocho extract according to the present invention.
도 5는 본 발명에 따른 신선초 추출물의 마우스 허파에서의 인터루킨-13 사이토카인 분비 억제 효과를 나타낸 사진이다.Figure 5 is a photograph showing the inhibitory effect of interleukin-13 cytokine secretion in mouse lungs of Sinchocho extract according to the present invention.
본 발명은 항천식 활성을 가지는 신선초(Angelica archangelica) 추출물에 관한 것으로서, 더욱 상세하게는, 신선초(Angelica archangelica)를 파쇄한 다음, C3-6의 저급알콜, 물, 저급유기산, 저급알콜에스테르, 저급케톤 및 할로겐화탄화수소로 구성된 군에서 선택된 용매로 추출한 항산화 및 항천식 활성을 가지는 신선초 추출물 및 그 제조방법에 대한 것이다. The present invention relates to an extract of Angelica archangelica having anti-asthma activity, and more particularly, after crushing Angelica archangelica , C 3-6 lower alcohol, water, lower organic acid, lower alcohol ester, It relates to a fresh vinegar extract having antioxidant and anti-asthmatic activity extracted with a solvent selected from the group consisting of lower ketones and halogenated hydrocarbons, and a method of preparing the same.
과민성 염증반응의 대표적인 질환으로서 천식 (asthma)을 들 수 있다. 이는 아토피성 피부염, 발진, 기관지 천식, 알레르기성 비염 등과 함께 중요한 알레르기 질환으로 알려져 있다. 최근 천식의 발병률은 급증하는 현상을 보이고 있는데, 이는 공업화에 따른 대기오염의 영향 등이 큰 것으로 알려져 있다. 천식은, 면역적인 과민 반응의 제반 증상(재치기, 콧물, 감기증세, 그리고 가려움증)외에 심지어 정맥동염 (sinusitis)이나 천식과 같은 심각한 호흡기 만성질환으로 연결되어 생명에 지장을 초래 할 수 있는 것으로 알려져 있다. Asthma is a representative disease of an overactive inflammatory response. It is known as an important allergic disease along with atopic dermatitis, rash, bronchial asthma and allergic rhinitis. Recently, the incidence of asthma has been rapidly increasing, which is known to be largely affected by air pollution due to industrialization. Asthma is known to lead to life-threatening chronic respiratory illnesses such as sinusitis and asthma, in addition to all the symptoms of immune hypersensitivity reactions (sickness, runny nose, cold symptoms, and itching). .
공해물질에서 나오는 산화스트레스(oxidant stress)는 대표적으로 과산화이온(superoxide anion (O2-))과 산화수소(hydro radical (OH-)) 등이 있는데, 이들 물질은 공유전자쌍을 가지지 않기 때문에 매우 불안정한 분자들이다. 생물학적으로 산화스트레스에 노출이 되면 전자전달계(마이토콘드리아의 전자전달계, 파고사이트의 활성)에 이상을 유발할 수 있는데, 이는 외부적인 요인에 의한 경우가 많다. 생물체 내에서는 허파나 산소의 2차 대사에 영향을 주게 되며, 최종적으로 이 는 단백질, 디엔에이(DNA), 지방 등의 대사에 영향을 준다(Lee, I., et al.. J. Biol. Chem. 280: 6094, 2005). Oxidative stress from the pollutant (oxidant stress) is typically a peroxide ion (superoxide anion (O 2-)) and the oxidation of hydrogen (hydro radical (OH -)) there is such, these materials are very unstable due to not have a shared pair of electrons Molecules. Biological exposure to oxidative stress can cause abnormalities in electron transport systems (such as mitochondrial electron transport systems and pagosite activity), which are often caused by external factors. In living organisms, it affects secondary metabolism of lungs and oxygen, which in turn affects metabolism of proteins, DNA, and fats (Lee, I., et al. J. Biol. Chem) 280: 6094, 2005).
산화스트레스는 천식과 밀접한 관련성을 가지고 있는 것으로 알려져 있는데, 그 대표적인 것이 산화스트레스에 의한 염증반응이다. 염증반응은 천식의 초기에 나타나는 대표적인 증상이다. 호흡기 질환 환자에 있어 산화제(oxidant)와 항산화제(anti-oxidant)의 비율이 정상인에 비해 불균형하게 나타나는 것으로 보고된 바 있는데, 치료의 면에서 수퍼옥사이드(superoxide)의 저해물질인 수퍼옥사이드디스무테이즈(superoxide dismutase (SOD))가 호흡기 질환을 완화시켜주는 것으로 알려져 있으며, 치료의 목적으로 항산화가 이용될 수 있다는 보고도 있다. 활성 산소(reactive oxygen species (ROS))는 면역계통에서도 이상을 유발하는 것으로 알려져 있어, 활성 산소를 억제하는 물질을 식품에 첨가하려는 보고도 많이 있다. 천식과 관련한 면역계통의 과민반응은 면역세포인 T-세포에 의한 경우가 많은데 이는 T-세포의 CD4 파지티브를 가지는 세포에서 분비하는 인터루킨-4와 인터루킨-13이라는 사이토카인과 관련성이 있다(Nadeem, A., et al.. J. Allergy. Clin. Immunol., 111: 72, 2003; MacNee, W. Eur. J. Pharmacol., 429: 195, 2001). Oxidative stress is known to be closely associated with asthma, a typical example of which is the inflammatory response caused by oxidative stress. Inflammatory reactions are a typical symptom of early asthma. In patients with respiratory diseases, the ratio of oxidant and anti-oxidant has been reported to be disproportionately compared to normal people, and superoxide dismutase, an inhibitor of superoxide, has been reported in terms of treatment. Superoxide dismutase (SOD) is known to alleviate respiratory diseases, and there are reports that antioxidants can be used for therapeutic purposes. Reactive oxygen species (ROS) are known to cause abnormalities in the immune system, and there are many reports on adding substances that inhibit active oxygen to foods. Asthma-related immune system hypersensitivity is most often caused by T-cells, which are immune cells, which are related to cytokines called interleukin-4 and interleukin-13, which are secreted by cells with CD4 positive T-cells (Nadeem). , A., et al. J. Allergy. Clin. Immunol. , 111: 72, 2003; MacNee, W. Eur. J. Pharmacol., 429: 195, 2001).
과산화수소(Hydrogen peroxide)는 세포막을 침투하여 세포의 신호전달과정에 이상을 일으키는 것으로 알려져 있다. 또한 과산화수소는 지질산화(lipid peroxidation)를 일으킬 뿐만 아니라 사이토졸릭 포스포리파아제 A2(cytosolic phospholipase A2 (cPLA2))와 프로스타글란딘 E2(prostaglandin E2 (PGE2))의 생산을 야기하게 된다. 여기서 프로스타글란딘 E2는 프로테인카이네이즈 C(protein kinase C (PKC)-mitogen activated protein kinase (MAPK) pathway)를 통한 자유라디칼억제제(free-radical scavengers)의 생산을 야기하게 된다. 이외에도 과산화수소는 스스로 신호전달자로서의 기능을 가지며, 상피세포성장인자(epidermal growth factor (EGF)) 같은 성장호르몬에도 영향을 미치는 것으로 알려져 있다(Bae, YS., et al.. J. Biol. Chem., 272: 217, 1997).Hydrogen peroxide is known to penetrate the cell membrane and cause abnormalities in cell signaling. Hydrogen peroxide not only causes lipid peroxidation but also causes the production of cytosolic phospholipase A2 (cPLA2) and prostaglandin E2 (PGE2). Prostaglandin E2 causes the production of free-radical scavengers through protein kinase C (PKC) -mitogen activated protein kinase (MAPK) pathway. In addition, hydrogen peroxide has a function as a signal transmitter itself, and is known to affect growth hormones such as epidermal growth factor (EGF) (Bae, YS., Et al. J. Biol. Chem., 272: 217, 1997).
현재, 천식에 대한 치료 방법은 그 질환의 원인이 아닌, 증세만 일시적으로 완화하려는 대증요법이 주를 이루고 있다. 즉, 기관지 천식이 발생하였을 경우에는 기관지 확장제를 사용하거나 항히스타민제를 사용하여 일시적으로 증세를 완화시켜 줄 뿐이며, 근원적인 예방 또는 치료방법이 알려져 있지 않다. Currently, the treatment for asthma is mainly symptomatic therapy to temporarily relieve only the symptoms, not the cause of the disease. In other words, when bronchial asthma occurs, the use of bronchodilators or antihistamines only temporarily relieves the symptoms, and there is no known preventive or therapeutic method.
이에, 천연물질로부터 각종 질환에 치료 효과가 있는 연구는 활발히 진행 중이나, 천식과 관련된 적절한 치료제는 아직 개발되지 않은 상태이다. Therefore, researches that are effective in treating various diseases from natural substances are actively conducted, but appropriate therapeutic agents related to asthma have not been developed yet.
한편, 신선초는 염증 치유, 경련해소, 담즙분비 촉진, 이뇨작용 등의 효능을 가진 식물로 학명은 Angelica archangelica이고, 원산지는 아시아와 유럽 지역이다. 신선초의 주요성분은 정유, 쿠마린, 비타민A,B1,B2~B12,비타민C,D, 엽록소 및 게르마늄이다. 신선초 추출물 중, 오일 성분은 췌장암세포(PANC-1 human pancreas cancer cells)에 독성을 가지는 것으로 알려져 있으며(Anticancer Res. 25(3B):1877, 2005), 췌장암 세포의 증식을 억제하며(Z Naturforsch, 59(7-8):523, 2004) 항암효과가 있는 것으로 알려져 있다(In Vivo. 19(1):191, 2005). On the other hand, it is a plant that has the effects of healing inflammation, relieving spasms, promoting bile secretion, and diuretic effect. The scientific name is Angelica archangelica , and its origin is Asia and Europe. The main components of fresh vinegar are essential oils, coumarins, vitamins A, B1, B2-B12, vitamin C, D, chlorophyll and germanium. Among the fresh vinegar extracts, the oil component is known to be toxic to PANC-1 human pancreas cancer cells ( Anticancer Res. 25 (3B): 1877, 2005) and inhibits the proliferation of pancreatic cancer cells ( Z Naturforsch , 59 (7-8): 523, 2004) and are known to have anticancer effects ( In Vivo . 19 (1): 191, 2005).
신선초의 부위별 추출물을 달리하여 생물학적 활성을 확인해 본 결과, 추출물은 클로로포룸(chloroform)을 이용하였을때, 추출부위는 뿌리에서 가징 큰 활성 을 보이는 것으로 알려졌다(Planta Med. 58(2):176, 1992). 이외에도 다양한 분석 방법 등을 이용하여 신선초의 추출물에 관한 연구가 보고되어 있다(Acta Pharm. 54(4):277, 2004; Eur J Histochem. 47(1):87, 2003). 특히 신선초는 소화 장애와 관절염 등에 예부터 이용이 되어 왔는데, 이는 신선초의 항산화 효과가 알코올로 유도된 간암마우스에 좋은 효과를 보이는 것으로 연구된 바 있다(Pharmacology. 68(2):70, 2003). As a result of confirming the biological activity by different extracts of fresh vinegar, it was found that the extract showed a large activity at the root when the extract was used chloroform ( Planta Med. 58 (2): 176, 1992). In addition, studies on extracts of fresh vinegar using various analysis methods have been reported ( Acta Pharm. 54 (4): 277, 2004; Eur J Histochem. 47 (1): 87, 2003). In particular, fresh vinegar has been used for a long time in digestive disorders and arthritis, and the antioxidant effect of fresh vinegar has been shown to have a good effect on alcohol-induced liver cancer mice ( Pharmacology. 68 (2): 70, 2003).
그러나, 상기한 바와 같이, 신선초는 다양한 질병에 탁월한 효과가 보고되고 있으나, 천식과 관련한 연구는 이뤄지지 않고 있는 실정이다. However, as described above, although the fresh vinegar has been reported to have excellent effects on various diseases, research on asthma has not been made.
이에, 본 발명자들은 천연물질에서 추출하여, 인체에 무해하고 천식을 억제할 수 있는 새로운 조성물을 개발하고자 예의 노력한 결과, 신선초 추출물이 산화스트레스 활성을 저해하고, 천식을 유의적으로 억제하여 우수한 항산화 및 항천식 효과를 나타냄을 확인하고, 본 발명을 완성하게 되었다. Accordingly, the present inventors have been eager to develop a new composition that is harmless to the human body by extracting from natural substances, as a result, the fresh vinegar extract inhibits the oxidative stress activity, significantly inhibits asthma and excellent antioxidant and It was confirmed that it exhibits an anti-asthma effect, and the present invention was completed.
결국, 본 발명의 목적은 항산화 및 항천식 효과를 갖는 신선초 추출물 및 이를 유효성분으로 함유하는 항산화 또는 천식의 치료를 위한 약학조성물을 제공하는데 있다. After all, it is an object of the present invention to provide a pharmaceutical composition for the treatment of antioxidant or asthma containing the fresh vinegar extract having antioxidant and anti-asthmatic effects as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 (a) 신선초(Angelica archangelica)를 파쇄한 다음, C3-6의 저급알콜, 물, 저급유기산, 저급알콜에스테르, 저급케톤 및 할로겐화탄화수소로 구성된 군에서 선택된 용매로 추출하는 단계; 및 (b) 상기 추출액을 회수하는 단계를 포함하는 항산화 및 항천식 활성을 가지는 신선초 추출물의 제조방법을 제공하며, 본 발명에 있어서, 상기 용매는 물인 것을 특징으로 할 수 있다.In order to achieve the above object, the present invention (a) crushed Angelica archangelica , and then selected from the group consisting of C 3-6 lower alcohol, water, lower organic acid, lower alcohol ester, lower ketone and halogenated hydrocarbon Extracting with a solvent; And (b) provides a method for producing a fresh vinegar extract having antioxidant and anti-asthmatic activity comprising the step of recovering the extract, in the present invention, the solvent may be characterized in that the water.
본 발명은 또한, 상기 방법에 의해 제조되고, 항산화 및 항천식 기능을 가지는 신선초(Angelica archangelica) 추출물을 제공한다.The present invention also provides an extract of Angelica archangelica prepared by the above method and having antioxidant and anti-asthmatic function.
본 발명에 있어서, 상기 신선초 추출물은 천식 유발 세포인 CD4 파지티브 T-세포를 억제하는 것을 특징으로 할 수 있고, 상기 신선초 추출물은 천식 유발 단백질인 인터루킨-13 또는 인터루킨-4의 발현을 억제하는 기능을 가지는 것을 특징으로 할 수 있다. In the present invention, the fresh vinegar extract may be characterized by inhibiting CD4 positive T-cells that are asthma-induced cells, the fresh vinegar extract is a function of inhibiting the expression of interleukin-13 or interleukin-4, which is an asthma-inducing protein It may be characterized by having.
본 발명은 또한, 상기 신선초 추출물을 유효성분으로 함유하는 항산화 또는 항천식용 약학 조성물을 제공하며, 상기 약학 조성물은 항히스타민제, 소염진통제, 항암제, 항생제로 이루어진 그룹으로부터 선택된 어느 하나 이상의 약제와 함께 제제화된 것을 특징으로 할 수 있다. The present invention also provides an antioxidant or anti-asthma pharmaceutical composition containing the fresh vinegar extract as an active ingredient, wherein the pharmaceutical composition is formulated with at least one agent selected from the group consisting of antihistamines, anti-inflammatory analgesics, anticancer agents, and antibiotics. It may be characterized as.
본 발명은 또한, 상기 신선초 추출물 및 식품학적으로 허용 가능한 식품보조 첨가제를 함유하는 항산화 또는 항천식용 건강기능성식품을 제공한다.The present invention also provides an antioxidant or anti-asthmatic health functional food containing the fresh vinegar extract and a food supplement acceptable food supplement.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 신선초 추출물은 다음과 같은 방법에 의해 제조될 수 있다. 본 발명의 신선초를 구매/수확하여 분말화한 후, 신선초 시료 중량의 약 2~10배에 달하는 부피의 물을 가하여 5~10시간 동안 열수추출, 냉침 추출, 환류냉각추출 또는 초음파추출 등의 추출방법을 사용하여 추출한 다음, 원심분리하여 신선초 추출물을 수득할 수 있다.Fresh vinegar extract of the present invention can be prepared by the following method. Purchasing / harvesting the fresh vinegar of the present invention into powder, and then adding water of a volume of about 2 to 10 times the weight of the fresh vinegar to extract water such as hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction for 5-10 hours. Extracted using the method, and then centrifuged to obtain a fresh vinegar extract.
본 발명의 신선초 추출물의 경우, 천연물질이므로 독성이 전혀 없어서 의약품으로 지속적으로 다량 사용할 수 있다. Fresh vinegar extract of the present invention, because it is a natural substance can be continuously used in large amounts as a drug because there is no toxicity.
상기의 제법으로 수득된 본 발명의 신선초 추출물을 과산화수소를 이용한 산화스트레스의 유발 및 이를 이용한 세포손상 억제 기법에 의하여 산화스트레스에 의한 세포 손상의 억제 활성을 확인한 결과, 탁월한 세포손상 억제 효과를 확인할 수 있었다. 또한, 자가면역체계를 조절하는 중요한 인자인 T-세포인 CD4 파지티브 세포의 발현 연상과 이에 의한 인터루킨-4 및 인터루킨-13의 분비를 억제하는 효과도 가지는 것을 확인할 수 있었다. 본 발명은 상기와 같이 산화스트레스에 의한 세포손상의 억제 및 천식관련 자가면역조절 활성을 갖는 신선초 추출물을 유효성분으로 함유하는 항산화 또는 천식 치료를 위한 약학 조성물을 제공한다.Induction of oxidative stress using hydrogen peroxide and inhibition of cell damage by oxidative stress using the peroxidation stress using the same as the extract of the fresh vinegar of the present invention obtained by the above method, the excellent cell damage inhibition effect was confirmed . In addition, it was confirmed that the T-cells, which are important factors for regulating the autoimmune system, are associated with the expression of CD4 positive cells and thereby inhibit the secretion of interleukin-4 and interleukin-13. The present invention provides a pharmaceutical composition for the treatment of antioxidant or asthma containing as an active ingredient a fresh vinegar extract having inhibition of cell damage by oxidative stress and asthma related autoimmune regulatory activity as described above.
본 발명의 신선초 추출물을 포함하는 조성물은 이미 사용되고 있는 항히스타민제, 소염진통제, 항암제 및 항생제 등의 약제와 함께 제제화하거나 병용하여 사용될 수 있다. The composition comprising the fresh vinegar extract of the present invention can be used in combination or formulated with drugs such as antihistamines, anti-inflammatory analgesics, anticancer agents and antibiotics that are already in use.
본 발명의 조성물의 약학적 투여 형태는 이들의 약학적 허용 가능한 염의 형태로도 사용될 수 있고, 또한 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아 니라 적당한 집합으로 사용될 수 있다.Pharmaceutical dosage forms of the compositions of the present invention may be used in the form of their pharmaceutically acceptable salts, or may be used alone or in combination with other pharmaceutically active compounds, as well as in a suitable collection.
본 발명에 따른 추출물을 포함하는 약학 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 추출물을 포함하는 조성물에 포함될 수 있는 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. Pharmaceutical compositions comprising extracts according to the invention, respectively, in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, external preparations, suppositories and sterile injectable solutions according to conventional methods. Can be formulated and used. Carriers, excipients and diluents that may be included in the composition comprising the extract include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.
제제화 할 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 추출물에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 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), 폴 리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents and surfactants are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose in the extract. ) Or lactose, gelatin and the like are mixed. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters 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.
본 발명의 신선초 추출물의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서, 본 발명의 추출물은 1일 0.0001 내지 100mg/kg으로, 바람직하게는 0.001 내지 100mg/kg으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the fresh herb extract of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and may be appropriately selected by those skilled in the art. However, for the desired effect, the extract of the present invention is preferably administered at 0.0001 to 100 mg / kg, preferably at 0.001 to 100 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
본 발명의 추출물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내 주사에 의해 투여될 수 있다. The extract of the present invention can be administered to mammals such as mice, mice, livestock, humans, etc. by various routes. All modes of administration can be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injections.
본 발명의 추출물을 함유하는 화합물 또는 약학적으로 허용 가능한 그의 염은 다양한 기능성 식품 및 건강보조식품의 제조시 식품의 주성분 또는 첨가제 및 보조제로 사용될 수 있다.The compound containing the extract of the present invention or a pharmaceutically acceptable salt thereof may be used as a main ingredient or additive and adjuvant of food in the preparation of various functional foods and dietary supplements.
본 명세서에서 '기능성 식품'이란, 일반 식품에 본 발명의 신선초 추출물을 첨가함으로써 일반 식품의 기능성을 향상시킨 식품을 의미한다. 기능성은 물성 및 생리기능성으로 대별될 수 있는데, 본 발명의 추출물을 일반식품에 첨가할 경우, 일반 식품의 물성 및 생리기능성이 향상될 것이고, 본 발명은 이러한 향상된 기능 의 식품을 포괄적으로 '기능성 식품'이라 정의한다.As used herein, the term "functional food" means a food product having improved functionality of a general food product by adding the fresh vinegar extract of the present invention to a general food product. Functionality can be roughly divided into physical properties and physiological functions. When the extract of the present invention is added to general foods, the physical properties and physiological functions of general foods will be improved, and the present invention comprehensively describes foods having such improved functions as 'functional foods'. 'Is defined.
예를 들어, 본 발명의 추출물의 항산화 및 항천식 효과를 이용하여 항천식 기능성식품을 제조할 수 있다. 또한 항천식 효과를 이용한 기능성 강화식품 등을 제조할 수 있을 것이다.For example, anti-asthma functional food can be prepared using the antioxidant and anti-asthma effects of the extract of the present invention. In addition, it will be possible to manufacture functional fortified foods using the anti-asthma effect.
상기 외에 본 발명의 추출물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 신선초 추출물은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 신선초 추출물 100중량부 당 0.01~20중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the extract of the present invention includes various nutrients, vitamins, minerals (electrolytes), flavors such as synthetic flavors and natural flavors, coloring and neutralizing agents (such as cheese and chocolate), pectic acid and salts thereof, alginic acid and its Salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the fresh vinegar extract of the present invention may contain a fruit flesh for the production of natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The ratio of such an additive is not so important, but it is generally selected in the range of 0.01 to 20 parts by weight per 100 parts by weight of fresh vinegar extract of the present invention.
이하, 실시예을 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지 않는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as limited by these examples.
실시예 1:Example 1: 신선초 추출물의 제조 Preparation of Sinchocho Extract
대구의 대형 유통업체에서 구입한 신선초를 경북대학교 생물학과 식물분류학 전문가에게 확인한 다음, 추출물 제조의 재료로 사용하였다. 막자사발 혹은 파쇄기를 이용하여 파쇄하고, 신선초 파쇄물 1g에 추출용매로 물을 10mL씩 첨가하여 5~10시간 동안 상온에서 추출하였으며, 추출 후 원심분리기를 이용하여 분당 5,000회전으로 30분간 원심분리하여 상등액을 회수하여 신선초 추출물을 수득하였다.Sinseoncho purchased from Daegu's major distributors was confirmed by Kyungpook National University's biology and plant taxonomy specialists, and used as an extract material. Crushed using a mortar or pestle, and extracted with 10 mL of water as an extractant to 1 g of fresh vinegar crushed product, extracted at room temperature for 5 to 10 hours. Was collected to obtain a fresh vinegar extract.
실시예 2: Example 2: 신선초 추출물의 산화스트레스 저해 활성 검색Screening of Oxidative Stress Inhibitory Activity of Extract
상기 실시예 1에서 수득한 신선초 추출물의 시험관내(in vitro) 과산화수소를 이용한 산화스트레스에 의한 세포손상 저해를 알아보기 위하여, 뉴로블라스토마 세포주(SH-SY5Y)를 이용하여 과산화수소(100μM)와 실시예 1에서 수득한 신선초 추출물 10μL (200μL 반응액 중)를 투여하여 현미경과 엠티티(MTT) 활성 측정을 이용하여 산화스트레스에 의한 세포손상 정도를 측정 하였다(도 1 및 도 2). In order to investigate the inhibition of cell damage by oxidative stress using in vitro hydrogen peroxide of the fresh vinegar extract obtained in Example 1, hydrogen peroxide (100 μM) and Examples using a neuroblastoma cell line (SH-SY5Y) 10 μL of fresh vinegar extract (in 200 μL reaction solution) obtained in 1 was administered, and the degree of cellular damage caused by oxidative stress was measured using a microscope and MTT activity measurement (FIGS. 1 and 2).
그 결과, 도 1에 나타난 바와 같이, 첫 번째 사진인, 산화스트레스를 받지 않은 세포(무첨가군)에서는 세포의 형태가 정상적으로 나타났지만, 두 번째 사진인, 산화스트레스를 받은 세포(100μM의 H2O2 첨가)는 산화스트레스로 인한 손상으로 세포막이 파괴된 것을 알 수 있었다. 그러나, 과산화수소 100μM과 실시예 1의 신선초 추출물을 10μL (200μL 반응액 중) 동시에 투여한 세포는 도 1의 세번째 그림과 같이 세포막이 정상적으로 유지가 되는 것을 알 수 있었다. As a result, as shown in Fig. 1, the cell morphology was normal in the first photo, the oxidative stress-free cells (no addition group), but the second photo, the oxidative stressed cells (100 μM H 2 O). 2 ) was found to damage cell membranes due to oxidative stress. However, it was found that the cell membranes were normally maintained as shown in the third image of FIG. 1, in which 100 μM of hydrogen peroxide and 10 μL (in 200 μL reaction solution) of the fresh vinegar extract of Example 1 were simultaneously administered.
또한, 엠티티(MTT) 활성을 측정한 결과, 도 2에 나타난 바와 같이, 뉴로블라 스토마 세포주(SH-SY5Y)에 과산화수소(100μM)만 첨가한 것보다, 과산화수소 100μM과 실시예 1의 신선초 추출물을 10μL (200μL 반응액 중) 동시에 투여한 세포주의 세포생존율은 약 40% 정도 증가하는 것을 알 수 있었다(도 2). In addition, as a result of measuring the activity (MTT), as shown in Figure 2, 100 μM hydrogen peroxide and fresh vinegar extract of Example 1 than the addition of hydrogen peroxide (100 μM) to neuroblastoma cell line (SH-SY5Y) The cell survival rate of 10 μL (in 200 μL reaction solution) at the same time increased about 40% (FIG. 2).
따라서, 본 발명의 신선초 추출물은 신화스트레스에 의한 세포손상 억제 활성이 매우 우수한 것을 확인할 수 있었다. Therefore, the fresh vinegar extract of the present invention was confirmed that the cell damage suppression activity by mythic stress is very excellent.
실시예 3:Example 3: 천식억제 활성을 확인하기 위한 동물모델 완성 Complete animal model to confirm asthma suppression
상기 실시예 1의 신선초 추출물의 천식저해 활성을 확인하기 위하여 마우스를 이용한 천식모델을 이용하였다.An asthma model using a mouse was used to confirm the asthma inhibitory activity of Sinchocho extract of Example 1.
마우스(C57BL/6, male, 6~8주령)의 호흡기에 오발부민(50μg/mL)을 약 2달간 매일 1회 투여하여 호흡기에 면역반응을 유도하였다. 신선초 추출물의 천식억제 효과를 알아보기 위하여 2주간 매일 1mL의 신선초 추출물을 경구투여 하였다. 이후 마우스의 허파를 절취하여 포르말린으로 고정 후 파라핀 블록을 만들어 5~10μm로 절편을 만들었다. Ovalbumin (50 μg / mL) was administered once daily for about two months to induce an immune response to the respiratory tract of mice (C57BL / 6, male, 6-8 weeks old). In order to investigate the asthma inhibitory effect of fresh vinegar extract, oral administration of 1 mL fresh vinegar extract daily for two weeks. Since the lungs of the mouse was cut and fixed with formalin, paraffin blocks were made and sections were made into 5-10 μm.
실시예 4:Example 4: 신선초 추출물에 의한 허파의 CD4 파지티브 세포 저해 활성 확인 Confirmation of CD4 Positive Cell Inhibition Activity of Lung by Fresh Extract
상기 실시예 3으로부터 만들어진 조직 절편을 이용하여 CD4 안티바디를 이용하여 CD4 파지티브 T-세포를 면역 염색을 수행하였다(도 3). 그 결과, 도 3에 나타난 바와 같이, 면역 염색 결과 천식이 유도된 마우스의 허파에(좌측사진)서는 CD4 안티바디에 의해 갈색으로 염색된 T-세포가 상당수 관찰이 됨을 알 수 있었으 며, 본 발명의 신선초 추출물을 투여한 마우스의 허파(우측사진)에서는 CD4 파지티브 T-세포가 나타나지 않았다. Tissue sections made from Example 3 were used for immunostaining CD4 positive T-cells using CD4 antibodies (FIG. 3). As a result, as shown in Figure 3, the immunostaining results showed that a large number of T-cells stained brown by CD4 antibody were observed in the lungs (left picture) of asthma-induced mice. In the lungs (right picture) of mice to which the fresh herb extract was administered, no CD4 positive T-cells were observed.
실시예 5:Example 5: 신선초 추출물에 의한 허파의 인터루킨-4의 저해 활성 확인 Inhibition of Interleukin-4 in Lung by Fresh Extract
상기 실시예 3으로부터 만들어진 조직 절편을 이용하여 인터루킨-4 안티바디를 이용하여 세포로부터 분비되는 인터루킨-4의 양을 면역 염색을 통해 알아보았다. 그 결과, 도 4에 나타난 바와 같이, 면역 염색 결과 천식이 유도된 마우스의 허파에(좌측사진)서는 인터루킨-4 항체에 의해 갈색으로 염색된 부분이 상당수 관찰이 됨을 알 수 있었으며, 본 발명의 신선초 추출물을 투여한 마우스의 허파(우측사진)에서는 인터루킨-4가 나타나지 않았다. The amount of interleukin-4 secreted from cells using interleukin-4 antibodies using the tissue sections prepared from Example 3 was examined by immunostaining. As a result, as shown in Figure 4, the immunostaining results in the lungs (left picture) of asthma-induced mice, it can be seen that a large number of brown stained by the interleukin-4 antibody was observed, the fresh vinegar of the present invention The lungs (right picture) of the mice receiving the extract did not show interleukin-4.
실시예 6:Example 6: 신선초 추출물에 의한 허파의 인터루킨-13의 저해 활성 확인 Inhibitory Activity of Interleukin-13 in Lung by Fresh Extract
상기 실시예 3로부터 만들어진 조직 절편을 이용하여 인터루킨-13 안티바디를 이용하여 세포로부터 분비되는 인터루킨-13의 양을 면역 염색을 통해 알아보았다. 그 결과, 도 5에 나타난 바와 같이, 면역 염색 결과 천식이 유도된 마우스의 허파(좌측사진)에서는 인터루킨-13 항체에 의해 갈색으로 염색된 부분이 상당수 관찰이 됨을 알 수 있었으며, 본 발명의 신선초 추출물을 투여한 마우스의 허파(우측사진)에서는 인터루킨-13이 나타나지 않았다.The amount of interleukin-13 secreted from the cells using the interleukin-13 antibody using the tissue sections prepared from Example 3 was examined by immunostaining. As a result, as shown in Figure 5, the immunostaining results in the lungs (left picture) of the mice induced asthma, it can be seen that a large number of the stained part of the brown stained by the interleukin-13 antibody, the fresh vinegar extract of the present invention In the lungs (right picture) of the mice to which administration was performed, no interleukin-13 was observed.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통 상의 지식을 가진 자에 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일뿐이며, 이에 의해 본 발명의 범위가 제하되는 것이 아닌 점은 명백 할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.As described above in detail specific parts of the present invention, those skilled in the art, these specific descriptions are only preferred embodiments, which does not limit the scope of the present invention by Will be obvious. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
이상에서 살펴본 바와 같이, 본 발명의 신선초 추출물은 과산화수소에 의한 산화스트레스를 억제하고, 천식 유발의 중요한 세포로 알려진 CD4 파지티브 T-세포를 억제하는 효과가 있다. 또한, 본 발명의 신선초 추출물은 천식 유발의 중요한 분자인 인터루킨-4, 인터루킨-13의 사이토카인의 분비를 억제하여, 천식을 유발하는 면역세포의 활성을 저해하여 탁월한 천식의 억제 효과를 나타내므로, 본 발명의 신선초 추출물을 포함하는 조성물은 자가면역세포의 활성화에 의해 야기되는 천식의 치료에 효과적인 의약품으로 이용될 수 있다. As described above, the fresh vinegar extract of the present invention has the effect of inhibiting the oxidative stress caused by hydrogen peroxide, and inhibits CD4 positive T-cell known as an important cell of asthma induction. In addition, the fresh vinegar extract of the present invention inhibits the secretion of cytokines of interleukin-4 and interleukin-13, which are important molecules for inducing asthma, inhibits the activity of immune cells causing asthma, and thus exhibits an excellent inhibitory effect on asthma. The composition comprising the fresh vinegar extract of the present invention can be used as an effective medicine for the treatment of asthma caused by the activation of autoimmune cells.
본 발명의 신선초 추출물은 천연물질에서 추출하여, 인체에 무해하여 항천식용 의약품으로 지속적인 사용이 가능하다. 또한 기존에는 천식의 근본적인 치료가 아닌 대증요법이 치료의 주종을 이루었으나, 본 발명의 신선초 추출물은 항산화 및 항천식 효과가 있어 천식의 근본적인 치료를 가능하게 한다. Fresh vinegar extract of the present invention is extracted from natural substances, harmless to the human body can be used continuously as an anti-asthma medicine. In addition, the conventional treatment of asthma, not the fundamental treatment of asthma, but the main dominance of the treatment, the extract of the present invention is an antioxidant and anti-asthma effect enables the fundamental treatment of asthma.
Claims (8)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
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KR1020060063589A KR100825869B1 (en) | 2006-07-06 | 2006-07-06 | Extract of Angelica archangelica Having Anti-Asthma Activity |
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KR1020060063589A KR100825869B1 (en) | 2006-07-06 | 2006-07-06 | Extract of Angelica archangelica Having Anti-Asthma Activity |
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