KR101505941B1 - Composition for preventing, improving or treating influenza virus infection and inflammatory disease comprising leek extract as an active ingredient - Google Patents
Composition for preventing, improving or treating influenza virus infection and inflammatory disease comprising leek extract as an active ingredient Download PDFInfo
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- KR101505941B1 KR101505941B1 KR1020130062628A KR20130062628A KR101505941B1 KR 101505941 B1 KR101505941 B1 KR 101505941B1 KR 1020130062628 A KR1020130062628 A KR 1020130062628A KR 20130062628 A KR20130062628 A KR 20130062628A KR 101505941 B1 KR101505941 B1 KR 101505941B1
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- influenza virus
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Abstract
본 발명은 대파 추출물의 항인플루엔자 바이러스 및 항염증 효과에 대한 것으로, 더욱 상세하게는 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 및 염증 예방 및 치료용 약학적 조성물 및 건강기능성 식품 조성물에 대한 것이다. 본 발명은 대파 추출물의 항바이러스 및 항염증 효과를 이용하여 인플루엔자 바이러스 감염 및 염증을 예방, 개선 및 치료하는데 탁월한 효과가 있다.The present invention relates to an anti-influenza virus and anti-inflammatory effect of a lupine extract, and more particularly to a pharmaceutical composition and a health functional food composition for influenza virus infection and inflammation prevention and treatment, which comprises a lupine extract as an active ingredient. The present invention has an excellent effect in preventing, ameliorating and treating influenza virus infection and inflammation by utilizing the antiviral and anti-inflammatory effects of the lobster extract.
Description
본 발명은 대파 추출물의 항인플루엔자 바이러스 및 항염증 효과에 대한 것으로, 더욱 상세하게는 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 및 염증성 질환 예방, 개선 및 치료용 조성물에 관한 것이다.The present invention relates to an anti-influenza virus and anti-inflammatory effect of a lupine extract, and more particularly, to a composition for preventing, ameliorating, and treating an influenza virus infection and an inflammatory disease, comprising an extract of lupine as an active ingredient.
인플루엔자는 orthomyxoviridae과 인플루엔자바이러스속에 속하는 바이러스로 호흡기질환을 일으킨다. 바이러스입자에는 피막이 있고, 지름 80~120nm의 구상이지만 사상형 입자 등 다형성을 띤다. 바이러스입자는 V항원, 즉 13종류의 적혈구응집소(HA) 및 9종류의 뉴라미다아제(NA)로 규정되는 표면항원과 S항원(주로 리보핵단백질 NP로 규정하는 항원)을 갖고 S항원의 혈청학적 특이성에 의해 A, B, C의 3형으로 나뉜다.A형은 돌연변이(drift)나 유전체분절교환(shift)에 의해 새로운 V항원(HA 또는 NA) 변이주가 출현하기 쉽고 이때 인간집단 속에서 변이항원에 대한 항체보유율이 낮으면 세계적으로 대유행을 일으킨다. 분리주의 명명은 형, 자연숙주, 아형, 분리지, 분리번호, 분리년도(HA 및 NA의 항원형)의 순서로 기재한다. 사람 외에 돼지, 말, 오리 등에서도 분리된다. 동물체 내에 서도 증식하고 분절교환이 일어난다. B형은 새로운 변이주가 출현하더라도 항원의 변이는 연속적이며, 유행은 주로 국지적이다. 원숭이, 소 등의 신장 및 태아배양세포로 증식하는 C형은 1개의 긴 유전체가 있고 표면항원의 변화는 가장 적다.
Influenza is a virus belonging to the genus orthomyxoviridae and the influenza virus, causing respiratory diseases. The virus particles have a coating, spherical shape with a diameter of 80 to 120 nm, but they have polymorphism such as filamentous particle. The viral particles have surface antigens defined by V antigens, namely 13 kinds of red blood cell aggregates (HA) and 9 kinds of neuramidases (NA), and S antigens (antigens mainly defined as ribonucleoprotein NPs) It is divided into three types of A, B and C by the serological specificity. A type A is susceptible to the appearance of a new V antigen (HA or NA) mutant by drift or genetic segmental shift, Low levels of antibody to mutant antigens cause pandemics worldwide. The name of the isolate should be written in the order of type, natural host, subtype, isolate, segregation number, segregation year (antinuclear form of HA and NA). In addition to people, pigs, horses, and ducks are also separated. Proliferation and segment exchange occur within the animal. In type B, even when a new mutant appears, the mutation of the antigen is continuous, and the epidemic is mainly local. Type C, which proliferates into kidney and fetal cultured cells such as monkeys and cows, has one long genome and the least change in surface antigen.
현재까지 개발된 항바이러스들은 심한 부작용을 나타내고 있으며 그 응용에 대한 많은 주의가 필요하다. 또한, 백신의 개발은 유행하는 바이러스의 형과 백신의 바이러스가 맞지 않으면 효과가 낮은 문제점이 있다.
The antiviruses developed so far show severe side effects and require a lot of attention to their application. In addition, development of a vaccine has a problem in that the effectiveness is low unless the virus type of the prevalent virus is matched with the virus of the vaccine.
따라서 감염 억제 효과가 뛰어나고 안정성이 우수한 새로운 인플루엔자 바이러스제의 개발의 필요성이 증가하고 있다.
Therefore, there is a growing need for the development of new influenza virus agents with excellent antiviral inhibition and excellent stability.
염증 반응은 손상이나 박테리아, 곰팡이, 바이러스 등 외부 물질에 의해 자극되어 각종 염증 매개 인자 및 면역 세포에 의한 효소의 활성화, 염증 매개물질 분비, 체액 침윤, 세포의 이동, 조직 파괴 등 일련의 복합적인 생리적 반응이 일어나는 것을 말하며, 이로 인해 홍반, 부종, 발열, 통증 등과 같은 증상이 수반된다. 염증 반응은 외부 감염원을 제거하고 손상된 조직을 재생하여 생명체의 기능을 회복시키는 작용을 하지만, 항원이 제거되지 않거나, 내부 물질이 원인이 되는 등 염증 반응이 과도하거나 지속적으로 일어나면 오히려 점막 손상, 조직 파괴 등이 일어나고, 암, 염증성 피부 질환, 관절염 등을 초래하기도 한다.
The inflammatory response is stimulated by foreign substances such as damage, bacteria, fungi and viruses, and a series of complex physiological phenomena such as activation of enzymes by various inflammatory mediators and immune cells, secretion of inflammatory mediators, fluid infiltration, cell migration, Reaction, which leads to symptoms such as erythema, edema, fever, and pain. Inflammation reaction removes external infectious agents and regenerates damaged tissues to restore the function of living organisms. However, if the inflammation reaction occurs excessively or continuously such as the antigen is not removed or the internal substance is caused, the mucous membrane damage, tissue destruction And cancer, inflammatory skin disease, arthritis, and others.
현재까지는 상기 염증성 질환의 치료를 위하여 주로 항히스타민제, 비타민 연고, 부신피질호르몬제가 사용되어왔으나, 이러한 약물은 그 효과가 일시적인 것이 대부분이고, 부작용이 심하여 염증성 질환의 치료 효과가 있는 새로운 물질의 개발이 요구되고 있다.Up to now, antihistamines, vitamin ointments, and corticosteroids have been mainly used for the treatment of inflammatory diseases. However, the development of new medicines that have a therapeutic effect on inflammatory diseases due to the serious side effects and most of them are temporary. Is required.
이에 본 발명자들은 항인플루엔자 바이러스 및 항염증 효능이 뛰어난 전통 재래 식품 추출물에 관하여 연구하던 중, 대파 추출물이 항인플루엔자 바이러스 및 항염증 활성이 뛰어나 인플루엔자 바이러스 감염 및 염증성 질환의 예방 및 치료하는데 효과가 있는 것을 발견하여 본 발명을 완성하였다.
Therefore, the inventors of the present invention have conducted studies on anti-influenza viruses and traditional traditional food extracts excellent in anti-inflammatory effect, and found that the extract of Daegwai had excellent anti-influenza virus and anti-inflammatory activity and was effective for the prevention and treatment of influenza virus infection and inflammatory diseases And completed the present invention.
따라서, 본 발명의 목적은 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 예방 및 치료용 약학적 조성물을 제공하는 것이다.
Accordingly, it is an object of the present invention to provide a pharmaceutical composition for preventing and treating influenza virus infection, which comprises an extract of Mallow Parlor as an active ingredient.
본 발명의 또 다른 목적은, 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 예방 및 개선용 건강기능성 식품 조성물을 제공하는 것이다.
It is still another object of the present invention to provide a health functional food composition for preventing and improving influenza virus infection, which comprises a lobster extract as an active ingredient.
본 발명의 또 다른 목적은, 대파 추출물을 유효 성분으로 포함하는 염증성 질환 예방 및 치료용 약학적 조성물을 제공하는 것이다.
It is still another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of inflammatory diseases, which comprises as an active ingredient an extract of Larval Larvae.
본 발명의 또 다른 목적은, 대파 추출물을 유효 성분으로 포함하는 염증성 질환 예방 및 개선용 건강기능성 식품 조성물을 제공하는 것이다.It is still another object of the present invention to provide a health functional food composition for preventing and improving inflammatory diseases, which comprises an extract of Lollandia as an active ingredient.
상기의 목적을 달성하기 위하여, 본 발명은 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 예방 및 치료용 약학적 조성물을 제공한다.
In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing and treating influenza virus infection comprising an extract of Mallow Parlor as an active ingredient.
본 발명의 다른 목적을 달성하기 위하여, 본 발명은 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 예방 및 개선용 건강기능성 식품 조성물을 제공한다.
In order to accomplish still another object of the present invention, the present invention provides a health functional food composition for preventing and improving influenza virus infection comprising an extract of Larval Root as an active ingredient.
본 발명의 다른 목적을 달성하기 위하여, 본 발명은 대파 추출물을 유효 성분으로 포함하는 염증성 질환 예방 및 치료용 약학적 조성물을 제공한다.
In order to accomplish still another object of the present invention, the present invention provides a pharmaceutical composition for the prevention and treatment of inflammatory diseases, which comprises an extract of Mallowia lupus as an active ingredient.
본 발명의 다른 목적을 달성하기 위하여, 본 발명은 대파 추출물을 유효 성분으로 포함하는 염증성 질환 예방 및 개선용 건강기능성 식품 조성물을 제공한다.
In order to accomplish another object of the present invention, the present invention provides a health functional food composition for prevention and improvement of inflammatory diseases comprising an extract of Lolium longan as an active ingredient.
이하 본 발명을 상세히 설명한다.
Hereinafter, the present invention will be described in detail.
본 발명의 약학적 조성물은 대파 추출물을 유효 성분으로 포함하여 인플루엔자 바이러스 감염 및 염증성 질환 예방 및 치료에 효과적이다.
The pharmaceutical composition of the present invention is effective for the prevention and treatment of influenza virus infection and inflammatory diseases by containing a lily extract as an active ingredient.
파는 중국의 서부에서 처음 생산된 향신채소다. 중국에서는 3천 년 전부터 재배하기 시작했고, 우리나라는 삼국시대 이전부터 재배한 기록이 있다. 현재는 동양의 온대지방에 걸쳐 재배가 이루어지는데 유럽이나 열대지방에는 그다지 널리 보급되지 않았다. 영양가는 흰 부분보다 파란 부분에 많은데 비타민A·C·K, 칼슘 등이 함유돼 있다.또한 몸을 따뜻하게 해, 열을 내리고 기침이나 담을 없애준다고 해서 감기의 특효 채소로 알려져 있기도 하다. 하얀 부분은 총백이라 하는데, 한방에서는 담 제거와 발한, 이뇨작용을 위해 그리고 구충약으로 이용한다. 파 달인 물은 류머티즘, 동상에 좋으며 신경안정과 피로회복 효과도 있다. 또한, 파 잎의 수가 많은 계통을 연화(軟化) 재배한 것을 대파 또는 움파라고 하며 노지에 재배하여 잎의 수가 적고 굵기가 가는 것을 실파라고 한다.
It is the first spice soda produced in western China. In China, cultivation began 3,000 years ago, and Korea has been cultivated since the Three Kingdoms period. Nowadays, cultivation takes place in the temperate regions of the East, but it has not spread widely in Europe and the tropics. Nutritional value is more in the blue part than in the white part, but it contains vitamins A, C, K, calcium, etc. It is also known as warming the body, lowering the heat, The whiteness is called "gangbang," which is used for desiccation and sweating, diuretic, and anthelmintics in one room. The paradent water is good for rheumatism, frostbite, and has nervous stability and fatigue recovery effect. In addition, the softened cultivated plant with a large number of leaf blades is called Daepa or Ummo, and it is grown in the field, and the number of leaves is small and the thickness is small.
본 발명의 대파 추출물은 공지의 천연물 추출방법에 의하여 추출될 수 있으며, 바람직하게는 물, 탄소수 1 내지 6개의 유기용매 및 아임계 또는 초임계 유체로 이루어진 군에서 선택된 하나 이상의 용매로 추출될 수 있으며, 상기 탄소수 1 내지 6개의 유기용매는 탄소수 1 내지 6개의 알코올(alcohol), 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌클로라이드(methylene chloride), 헥산(hexane), 시클로헥산(cyclohexane) 및 석유에테르(petroleum ether)로 이루어진 군 중에서 선택된 것일 수 있다.
The large intestine extract of the present invention can be extracted by a known natural substance extraction method and is preferably extracted with at least one solvent selected from the group consisting of water, organic solvents having 1 to 6 carbon atoms, and subcritical or supercritical fluids. , The organic solvent having 1 to 6 carbon atoms may be selected from the group consisting of alcohols having 1 to 6 carbon atoms, acetone, ether, benzene, chloroform, ethyl acetate, methylene chloride methylene chloride, hexane, cyclohexane, and petroleum ether.
본 발명의 추출물 추출온도는 특별히 제한되지 아니하며, 예를 들어 0℃ 내지 90 ℃일 수 있으며, 바람직하게는 상온 내지 50 ℃일 수 있다. 본 발명의 추출물 추출시간은 특별히 제한되지 아니하며, 예를 들어 1시간 내지 10일일 수 있으며, 바람직하게는 10시간 내지 24시간 일 수 있다.
The extraction temperature of the extract of the present invention is not particularly limited and may be, for example, from 0 ° C to 90 ° C, preferably from room temperature to 50 ° C. The extraction time of the extract of the present invention is not particularly limited, and may be, for example, 1 hour to 10 days, preferably 10 hours to 24 hours.
본 발명 추출물은 공지의 천연물 추출법으로 추출될 수 있다. 예를 들어 냉침추출, 열수추출, 초음파 추출, 환류냉각 추출, 가열 추출법으로 추출할 수 있으며, 바람직하게는 열수추출 또는 환류 냉각 추출법으로 추출할 수 있다.
The extract of the present invention can be extracted by a known natural substance extraction method. For example, it can be extracted by cold extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction or heat extraction, preferably by hot water extraction or reflux cooling extraction.
본 발명의 추출물은 여과하여 액상으로 사용할 수 있으며, 바람직하게는 분무건조 또는 동결건조의 건조 공정을 통하여 고형화하여 사용할 수 있다. 더욱 바람직하게는 분무건조 또는 동결건조 전 덱스트린을 혼합하여 건조할 수 있다.
The extract of the present invention can be used as a liquid phase by filtration, and can be solidified through a drying process such as spray drying or lyophilization. More preferably, the dextrin can be mixed and dried before spray drying or freeze-drying.
인플루엔자는 orthomyxoviridae과 인플루엔자바이러스속에 속하는 바이러스로 호흡기질환을 일으킨다. 인플루엔자 바이러스입자에는 피막이 있고, 지름 80~120nm의 구상이지만 사상형 입자 등 다형성을 띤다. 바이러스입자는 V항원, 즉 13종류의 적혈구응집소(HA) 및 9종류의 뉴라미다아제(NA)로 규정되는 표면항원과 S항원(주로 리보핵단백질 NP로 규정하는 항원)을 갖고 S항원의 혈청학적 특이성에 의해 A, B, C의 3형으로 나뉜다.A형은 돌연변이(drift)나 유전체분절교환(shift)에 의해 새로운 V항원(HA 또는 NA) 변이주가 출현하기 쉽고 이때 인간집단 속에서 변이항원에 대한 항체보유율이 낮으면 세계적으로 대유행을 일으킨다. 사람 외에 돼지, 말, 오리 등에서도 분리되며, 동물체 내에 서도 증식하고 분절교환이 일어난다. B형은 새로운 변이주가 출현하더라도 항원의 변이는 연속적이며, 유행은 주로 국지적이다. 원숭이, 소 등의 신장 및 태아배양세포로 증식하는 C형은 1개의 긴 유전체가 있고 표면항원의 변화는 가장 적다.
Influenza is a virus belonging to the genus orthomyxoviridae and the influenza virus, causing respiratory diseases. Influenza virus particles have a coating and spherical shape with a diameter of 80 to 120 nm, but they have polymorphism such as filamentous particle. The viral particles have surface antigens defined by V antigens, namely 13 kinds of red blood cell aggregates (HA) and 9 kinds of neuramidases (NA), and S antigens (antigens mainly defined as ribonucleoprotein NPs) It is divided into three types of A, B and C by the serological specificity. A type A is susceptible to the appearance of a new V antigen (HA or NA) mutant by drift or genetic segmental shift, Low levels of antibody to mutant antigens cause pandemics worldwide. In addition to humans, pigs, horses, and ducks are also isolated, and proliferation and segment exchange occur within the organism. In type B, even when a new mutant appears, the mutation of the antigen is continuous, and the epidemic is mainly local. Type C, which proliferates into kidney and fetal cultured cells such as monkeys and cows, has one long genome and the least change in surface antigen.
본 발명의 인플루엔자 바이러스는 바람직하게는 A형 인플루엔자일 수 있으며, 더욱 바람직하게는 A형 인플루엔자의 아형 중의 하나인 H1N1 바이러스 일 수 있다.
The influenza virus of the present invention may be preferably an influenza A virus, more preferably a H1N1 virus which is one of the subtypes of influenza A virus.
본 발명의 인플루엔자 바이러스 감염으로 인한 질환은 독감, 감기, 인후염, 기관지염, 폐렴, 발열, 오한, 두통 및 기침, 인후통, 콧물, 호흡곤란 등의 상기도증상, 근육통, 관절통, 피로감, 구토 및 설사일 수 있으나, 바람직하게는 독감, 감기, 인후염, 기관지염 및 폐렴일 수 있다.
The disease caused by the influenza virus infection of the present invention may be influenza symptoms such as flu, cold, sore throat, bronchitis, pneumonia, fever, chills, headache and cough, sore throat, runny nose, shortness of breath, muscular pain, joint pain, fatigue, But can be preferably flu, cold, sore throat, bronchitis and pneumonia.
본 발명의 일실시예에서는 대파 추출물의 농도별 인플루엔자 바이러스 억제 효과를 세포독성과 함께 실험하였다. 그 결과 대파 추출물은 농도 0.3704 mg/mL 이하에서는 세포독성이 낮으며, 0.0046 mg/mL에서는 항인플루엔자 바이러스 효과가 탁월한 것으로 나타났다(도 1 참조).
In one embodiment of the present invention, the inhibitory effect of influenza virus on the concentration of L. extract was tested together with cytotoxicity. As a result, it was found that the anti-influenza virus effect was excellent at a concentration of 0.3704 mg / mL or less and 0.0046 mg / mL, respectively.
한편 본 발명의 염증성 질환은 이에 제한되지는 않으나, 부종 등과 같은 일반적인 염증 증상을 포함하여 염증성 장 질환, 복막염, 골수염, 봉소염, 췌장염, 외상 유발 쇼크, 기관지 천식, 알러지성 비염, 낭포성 섬유증, 급성 기관지염, 만성 기관지염, 급성 세기관지염, 만성 세기관지염, 골관절염, 통풍, 척추관절병증, 강직성 척추염, 라이터 증후군, 건선성 관절병증, 장질환 척추염, 연소자성 관절병증, 연소자성 강직성 척추염, 반응성 관절병증, 감염성 관절염, 후-감염성 관절염, 임균성 관절염, 결핵성 관절염, 바이러스성 관절염, 진균성 관절염, 매독성 관절염, 라임 병, '혈관염 증후군'과 관련된 관절염, 결절성 다발동맥염, 과민성 혈관염, 루게릭 육아종증, 류마티스성 다발성근육통, 관절 세포 동맥염, 칼슘 결정 침착 관절병증, 가성 통풍, 비-관절 류마티즘, 점액낭염, 건초염, 상과염(테니스 엘보), 신경병증성 관절 질환(neuropathic joint disease; 또는 'charcot joint'이라고도 함), 출혈성 관절증(hemarthrosic), 헤노흐-쉔라인 자반병, 비후성 골관절병증, 다중심성 세망조직구종, 척추측만증(scoliosis), 혈색소증, 혈색소병증, 고지단백혈증, 저감마글로불린혈증, 가족성 지중해열(familial Mediterranean fever), 베체트 병, 전신성 홍반성 루푸스, 재귀열, 다발성 경화증, 패혈증, 패혈성 쇼크, 급성 호흡곤란 증후군, 다발성 장기부전, 만성 폐쇄성 폐질환(chronic obstructive pulmonary disease), 류마티스성 관절염(rheumatoid arthritis), 급성 폐손상(acute lung injury), 기관지 폐 형성장애(broncho-pulmonary dysplasia), 제2형 당뇨병, 동맥경화, 알츠하이머성 치매, 가족성 한랭 자가염증 증훈군(familial cold autoinflammatory syndrome), 머클-웰스 증후군(Muckle-Wells syndrome), 신생아발병다발염증성질환(neonatal mutisystem inflammatory disease), 만성유아신경피부관절증후군(chronic infantile neurologic cutaneous articular syndrome), 성인발증형 스틸병(adult-onset Still's disease), 접촉성 피부염, 포상기태(hydatidiform mole), PAPA 증후군(syndrome of pyogenic arthritis, pyoderma gangrenosum,and acne), 고면역글로불린 D 증후군(hyperimmunoglobulin d syndrome) 및 크리오피린 관련 주기적 증후군(cryopyrin-associated periodic syndromes)등을 포함할 수 있다.
The inflammatory diseases of the present invention include, but are not limited to, inflammatory bowel disease, peritonitis, osteomyelitis, cachexia, pancreatitis, trauma-induced shock, bronchial asthma, allergic rhinitis, cystic fibrosis, acute Inflammatory bowel disease, chronic bronchitis, acute bronchiolitis, chronic bronchitis, osteoarthritis, gout, spondyloarthropathies, ankylosing spondylitis, lager syndrome, psoriatic arthropathy, intestinal disease, inflammatory arthropathy, ankylosing spondylitis, reactive arthropathy, infectious arthritis , Arthritis associated with post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, viral arthritis, fungal arthritis, methotrexate arthritis, Lyme disease, vasculitis syndrome, nodular polyarteritis, irritable vasculitis, rheulic granulomatosis, rheumatoid polyposis muscle pain , Arthritic cell arthritis, calcific calcified arthropathy, caustic gout, non-joint Neuropathic joint disease (also referred to as "charcot joint"), hemarthrosis, Henoch-Schlenstein purpura, hypertrophic osteoarthritis, multiple sclerosis, multiple sclerosis, multiple sclerosis, Atherosclerosis, multiple sclerosis, multiple sclerosis, multiple sclerosis, scleroderma, scoliosis, hemochromatosis, hemoglobinopathy, hyperlipoproteinemia, hypogammaglobulinemia, familial Mediterranean fever, Behcet's disease, systemic lupus erythematosus, Acute respiratory distress syndrome, multiple organ failure, chronic obstructive pulmonary disease, rheumatoid arthritis, acute lung injury, broncho-pulmonary dysplasia ), Type 2 diabetes, arteriosclerosis, Alzheimer ' s dementia, familial cold autoinflammatory syndrome, Muckle-W syndrome neurological and neurological diseases, adult-onset Still's disease, contact dermatitis, reperfusion injury, neonatal inflammatory disease, hydatidiform mole, PAPA syndrome, pyoderma gangrenosum, and acne, hyperimmunoglobulin d syndrome, and cryopyrin-associated periodic syndromes.
본 발명의 일실시예에서는 대파 추출물의 항염증 효과를 알아보기 위해 염증조절복합체인 인플라마좀(inflammasome) 활성 억제시험을 하였다. 그 결과, 본 발명의 대파 추출물은 농도가 0.625mg/mL 이상 일때 인플라마좀을 효과적으로 억제하는 것으로 나타났다(도 3 , 도 4 및 도 5 참조).
In one embodiment of the present invention, inflammation-suppressing complex inflammation test was conducted to examine the anti-inflammatory effect of the extract of Lycopersicon esculentum. As a result, the L. extract of the present invention effectively inhibited inflammase when the concentration was 0.625 mg / mL or more (see FIGS. 3, 4 and 5).
인플라마좀은 골수성세포 (Myeloid cells)의 세포질에 존재하며 선천적면역수용체 (NOD-like receptor), 어뎁터 단백질 (ASC) 및 caspase-1로 이루어진 단백질복합체로써 염증반응에 관여하는 사이토카인의 전구체인 pro-Interleukin-1β (pro-IL-1β)를 활성형인 IL-1β (p17)으로 전환 시키는데 관여한다. 본 발명의 대파 추출물은 이러한 인플라마좀의 활성을 억제함으로써, 염증성 사이토카인 IL-1β의 분비 양을 효과적으로 조절할 수 있어 염증성 질환의 예방 및 치료에 탁월한 효과를 갖는다.
Inflammase is present in the cytoplasm of myeloid cells and is a protein complex composed of innate immune receptors (NOD-like receptor), adapter protein (ASC) and caspase-1. It is a precursor of cytokine -Interleukin-1? (Pro-IL-1?) Into the active form IL-1? (P17). The large-leaved extract of the present invention can effectively control the secretion amount of the inflammatory cytokine IL-1 beta by inhibiting the activity of this inflammase, and thus has an excellent effect for the prevention and treatment of inflammatory diseases.
따라서 대파 추출물을 유효성분으로 포함하는 본 발명의 약학적 조성물은 인플루엔자 바이러스 감염으로 인한 질환의 예방 및 치료에 효과적이며, 염증으로 인한 염증성 질환의 예방 및 치료에도 효과적이다.
Therefore, the pharmaceutical composition of the present invention comprising the extract of Robinia pseudo-acacia L. as an active ingredient is effective for prevention and treatment of diseases caused by influenza virus infection, and is also effective for prevention and treatment of inflammatory diseases caused by inflammation.
본 발명에 따른 약학적 조성물은 약학적으로 유효한 양의 대파 추출물을 단독으로 포함하거나 하나 이상의 약학적으로 허용되는 담체, 부형제 또는 희석제를 추가로 포함할 수 있다. 상기에서 "약학적으로 허용되는" 이란 생리학적으로 허용되고 인간에게 투여될 때, 활성성분의 작용을 저해하지 않으며 통상적으로 위장 장애, 현기증과 같은 알레르기 반응 또는 이와 유사한 반응을 일으키지 않는 비독성의 조성물을 말한다.
The pharmaceutical compositions according to the present invention may further comprise a pharmaceutically effective amount of the lice extract or may further comprise one or more pharmaceutically acceptable carriers, excipients or diluents. The term "pharmaceutically acceptable" as used herein means a non-toxic composition which is physiologically acceptable and which, when administered to humans, does not inhibit the action of the active ingredient and does not normally cause an allergic reaction such as gastrointestinal disorder, dizziness, .
상기 담체, 부형제 및 희석제의 예로는, 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸하이드록시벤조에이트, 프로필하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 또한 상기 약학 조성물은 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다.
Examples of the carrier, excipient and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition may further include a filler, an anti-coagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifier, and an antiseptic agent.
상기에서 "약학적으로 유효한 양"이란 음성 대조군에 비해 그 이상의 반응을 나타내는 양을 말하며 바람직하게는 인플루엔자 바이러스에 의해 유발되는 질환 또는 증상 예방 및 치료의 효과를 나타내기에 충분한 양을 말한다. 본 발명에 따른 대파 추출물의 약학적으로 유효한 양으로는 0.01 내지 100 mg/day/kg 체중이다. 그러나 상기 약학적으로 유효한 양은 질환의 종류 및 이의 중증정도, 환자의 연령, 체중, 건강상태, 성별, 투여 경로 및 치료기간 등과 같은 여러 인자에 따라 적절히 변화될 수 있다.
The term "pharmaceutically effective amount " as used herein refers to an amount that exhibits a reaction higher than that of the negative control, and preferably refers to an amount sufficient to exhibit the effect of preventing or treating a disease or symptom caused by influenza virus. The pharmacologically effective amount of the L. extract according to the present invention is 0.01 to 100 mg / day / kg body weight. However, the pharmaceutically effective amount may be appropriately changed depending on various factors such as the type and severity of the disease, the age, body weight, health condition, sex, administration route and treatment period of the patient.
또한, 본 발명의 약학적 조성물은 포유동물에 투여된 후 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 공지된 방법을 사용하여 제형화될 수 있다. 제형은 분말, 과립, 정제, 에멀젼, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캡슐, 멸균 주사용액, 멸균 분말의 형태일 수 있다.
In addition, the pharmaceutical composition of the present invention may be formulated using methods known in the art so as to provide rapid, sustained or delayed release of the active ingredient after administration to the mammal. The formulations may be in the form of powders, granules, tablets, emulsions, syrups, aerosols, soft or hard gelatine capsules, sterile injectable solutions, sterile powders.
본 발명 조성물의 투여 경로로는 이에 한정되지는 않으나 경구적 또는 비경구적으로 투여 될 수 있다. 비경구적 투여 경로로는 예를 들면, 경피, 비강, 복강, 근육, 피하 또는 정맥 등의 여러 경로가 포함된다.
The administration route of the composition of the present invention is not limited thereto, but it can be administered orally or parenterally. Parenteral routes of administration include, for example, various routes such as transdermal, nasal, peritoneal, muscular, subcutaneous or intravenous.
본 발명의 약학적 조성물은 인플루엔자 바이러스 감염으로 인한 질환 및 염증성 질환의 예방 및 치료의 효과를 가지는 공지의 화합물과 병행하여 투여할 수 있다.
The pharmaceutical composition of the present invention can be administered in combination with known compounds having the effect of preventing and treating diseases caused by influenza virus infection and inflammatory diseases.
한편 본 발명은 대파 추출물을 유효성분으로 포함하는 인플루엔자 바이러스 감염 예방 및 개선용 건강 기능성 식품 조성물을 제공한다.
The present invention also provides a health functional food composition for preventing and ameliorating influenza virus infection comprising an extract of Lollandia as an active ingredient.
본 발명의 건강 기능성 식품 조성물은 대파 추출물을 유효성분으로 포함하여 인플루엔자 바이러스 감염 예방 및 개선에 효과적이다.
The health functional food composition of the present invention is effective for prevention and improvement of influenza virus infection by containing a lily extract as an active ingredient.
또한, 본 발명은 대파 추출물을 유효성분으로 포함하는 염증성 질환 예방 및 개선용 건강 기능성 식품 조성물을 제공한다.
In addition, the present invention provides a health functional food composition for preventing and improving inflammatory diseases, which comprises as an active ingredient a lupine extract.
본 발명의 건강 기능성 식품 조성물은 대파 추출물을 유효성분으로 포함하여 염증성 질환 예방 및 개선에 효과적이다.
The health functional food composition of the present invention is effective for prevention and improvement of inflammatory diseases by containing an extract of Daechi wave as an active ingredient.
본 발명의 "건강기능식품"은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.
The term " health functional food "of the present invention means a food prepared and processed by using a raw material or ingredient having useful functionality in the human body according to Law No. 6727 on health functional foods, and" And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.
본 발명의 식품조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food) 및 식품 첨가제 (food additives) 등의 모든 형태를 포함한다. 상기 유형의 식품 조성물은 당 업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다.
The food composition of the present invention includes all forms such as functional food, nutritional supplement, health food and food additives. Food compositions of this type may be prepared in a variety of forms according to conventional methods known in the art.
예를 들면, 건강식품으로는 대파 추출물 자체를 차,주스 및 드링크의 형태로 제조하여 음용하도록 하거나, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다. 또한, 본 발명의 대파 추출물과 염증 및 인플루엔자 바이러스 감염에 의해 유발되는 질환 또는 증상의 예방 또는 개선의 효과가 있다고 알려진 공지의 활성 성분과 함께 혼합하여 조성물의 형태로 제조할 수 있다.
For example, the health food may be prepared by ingesting the lobster extract itself in the form of tea, juice and drink, or ingested by granulating, encapsulating and pulverizing. In addition, it may be prepared in the form of a composition by mixing together the active ingredient of the present invention with a known active ingredient known to be effective for preventing or ameliorating a disease or symptom caused by inflammatory and influenza virus infection.
또한, 기능성 식품으로는 음료(알콜성 음료 포함), 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마아말레이드 등), 어류, 육류 및 그 가공식품(예: 햄, 소시지 콘비이프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게티, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품(예: 버터, 치즈 등), 식용식물유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등) 등에 대파 추출물을 첨가하여 제조할 수 있다. 또한, 본 발명의 대파 추출물을 식품 첨가제의 형태로 사용하기 위해서는 분말 또는 농축액 형태로 제조하여 사용할 수 있다.
Functional foods also include beverages (including alcoholic beverages), fruits and their processed foods (e.g., canned fruits, bottled, jam, maalmalade, etc.), fish, meat and processed foods such as ham, Etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), fruit juice, various drinks, cookies, Retort foods, frozen foods, various seasonings (eg, soybean paste, soy sauce, sauces, etc.), etc. In addition, in order to use the L. extract of the present invention in the form of a food additive, it may be prepared in the form of powder or concentrate.
본 발명의 식품 조성물 중 대파 추출물의 바람직한 함량은 식품 조성물 총 중량에 대하여 0.001 내지 50 중량%일 수 있으며, 바람직하게는 0.1 내지 30 중량% 범위로 함유될 수 있다.The preferred content of the Greater Linear Extract in the food composition of the present invention may be 0.001 to 50% by weight, preferably 0.1 to 30% by weight, based on the total weight of the food composition.
본 발명은 대파 추출물의 항인플루엔자 바이러스 및 항염증 효과에 대한 것으로, 더욱 상세하게는 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 및 염증성 질환 예방 및 치료용 조성물에 관한 것이다. 본 발명은 대파 추출물의 항바이러스 효과를 이용하여 인플루엔자 바이러스 감염을 예방, 개선 및 치료에 효과적이며 또한 항염증 효과를 이용하여 염증성 질환을 예방, 개선 및 치료하는데 효과적이다.The present invention relates to an anti-influenza virus and anti-inflammatory effect of a lupine extract, and more particularly, to a composition for preventing and treating an influenza virus infection and an inflammatory disease, which comprises a lupine extract as an active ingredient. The present invention is effective in preventing, ameliorating, and treating influenza virus infection by using the antiviral effect of the lobster extract, and is effective in preventing, ameliorating and treating an inflammatory disease by using an anti-inflammatory effect.
도 1은 대파 추출물의 농도별 인플루엔자 억제효능을 타나낸 것이다. (가로: 대파 추출물의 고형물의 농도(mg/ml) , 세로: 세포활성도(O.D), con - : 인플루엔자 바이러스 없음, con + : 인플루엔자 바이러스 포함, cytotoxic : 대파 추출물의 세포독성, antiviral activity : 인플루엔자 억제효능)
도 2는 타미플루의 농도별 인플루엔자 억제효능을 나타낸 것이다. (가로축: 타미플루 농도(mg/mL), 세로축: 세포활성도(O.D) , con - : 인플루엔자 바이러스 없음, con + : 인플루엔자 바이러스 포함, cytotoxic : 타미플루의 세포독성, antiviral activity : 인플루엔자 억제효능))
도 3은 대파 추출물의 인플라마좀 억제 효능을 나타낸 것으로, 인플라마좀 억제 효과는 세포 배양으로 배출된 활성형 IL-1β의 양을 western blot으로 검출하였다. (① Sup. : 세포 배양액 , ② Lys. :세포내 단백질 추출물 , ③ IL-1β : 활성형 사이토카인, ④ pro-IL-1β : 전구체 사이토카인.) 이때, LPS->Non은 ATP를 처리하지 않은 negative control을 의미한다.
도 4는 대파 추출물의 농도(0mg/mL 내지 10mg/mL)에 따른, 도3의 세포배양액(Sup.)에서 검출된 IL-1β (활성형)의 양을 수치화한 그래프이다. 그래프의 검은색 막대는 ATP단독처리를 의미하며, 흰색 막대는 ATP와 대파 추출물을 복합투여했다는 의미이다. (가로: 대파 추출물의 농도(mg/ml), 세로: relative intensity(%))
도 5는 대파 추출물의 농도(0mg/mL 내지 2.5mg/mL)에 따른, 도3의 세포배양액(Sup.)에서 검출된 IL-1β (활성형)의 양을 수치화한 그래프이다. 그래프의 검은색 막대는 ATP단독처리를 의미하며, 흰색 막대는 ATP와 대파 추출물을 복합투여했다는 의미이다. (가로: 대파 추출물의 농도(mg/ml), 세로: relative intensity(%))Fig. 1 shows influenza inhibitory activity by the concentration of lobster extract. Con: Influenza virus free, con +: Influenza virus included, cytotoxic: Cytotoxic effect of lavash extract: Antiviral activity: Influenza inhibition efficacy)
Fig. 2 shows the effect of inhibiting influenza by the concentration of Tamiflu. (Influenza-inhibiting effect), (cytotoxic: cytotoxic effect of Tamiflu), (concentration of influenza virus)
FIG. 3 shows the effect of inhibiting the inflammation of the lobule extract. The effect of inhibiting the inflammosis was detected by western blotting the amount of active IL-1β released into the cell culture. (1) Sup.: Cell culture, (2) Lys.: Intracellular protein extract, (3) IL-1β: active cytokine, (4) pro-IL-1β: precursor cytokine. Negative control.
4 is a graph showing the quantification of the amount of IL-1? (Active) detected in the cell culture supernatant (Sup.) Of Fig. 3 according to the concentration (0 mg / mL to 10 mg / mL) The black bars in the graph mean ATP treatment alone, while the white bars indicate the combination of ATP and L. extract. (Width: concentration of lobster extract (mg / ml), height: relative intensity (%))
5 is a graph showing the quantification of the amount of IL-1? (Active) detected in the cell culture supernatant (Sup.) Of Fig. 3 according to the concentration (0 mg / mL to 2.5 mg / mL) The black bars in the graph mean ATP treatment alone, while the white bars indicate the combination of ATP and L. extract. (Width: concentration of lobster extract (mg / ml), height: relative intensity (%))
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
단, 하기 실시예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예에 한정되는 것은 아니다.
However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.
<실시예 1>≪ Example 1 >
대파 추출물 제조Manufacture of lobster extract
신선한 대파(Allium fistulosum, 영문 Leek)를 구입하여 뿌리를 절단하고 물로 세척하여 흙을 제거하고, 줄기를 사용하였다. 대파 줄기 1 kg에 에탄올(99.99% v/v)를 1,000 L 가하여 잘 혼합하고 정치하여, 실온에서 하룻밤 추출하였다. 거친 여과지(Advantec No. 1)을 사용하여 여과하여, 여액을 모아서 55~65 ℃에서 감압농축하였다. 에탄올이 완전히 제거되면 농축액을 모아서 미세 여과지 (Advantec No. 2)로 여과하였다. 여액을 모아 pH를 측정한 후, 1.0 또는 0.1 mol/L의 수산화나트륨 용액(NaOH)을 사용하여 pH 7.4로 조절하였다. pH가 조정된 여액의 일부를 취하여 105℃ 증발건조법을 사용하여 고형물함량을 측정한 후, 증류수를 추가하여 대파추출물의 최종 고형물 함량이 100 mg/mL로 조정하였다.
Fresh roe (Allium fistulosum, Leek) was purchased, the roots were cut, washed with water to remove soil, and stems were used. 1,000 L of ethanol (99.99% v / v) was added to 1 kg of Streptomyces stalks, mixed well and allowed to stand overnight at room temperature. The filtrate was collected using a coarse filter paper (Advantec No. 1), and the filtrate was collected and concentrated under reduced pressure at 55 to 65 ° C. When the ethanol was completely removed, the concentrate was collected and filtered through a micro filter paper (Advantec No. 2). The pH of the filtrate was collected and adjusted to pH 7.4 using 1.0 or 0.1 mol / L sodium hydroxide solution (NaOH). A portion of the pH-adjusted filtrate was taken and the solids content was measured using the evaporative drying method at 105 ° C., and the final solids content of the larvae extract was adjusted to 100 mg / mL by addition of distilled water.
<실시예 2>≪ Example 2 >
대파 추출물의 세포독성 (cytotoxicity) 및 인플루엔자 바이러스 억제효과Cytotoxicity and Influenza Virus Inhibitory Effect of Larval Extracts
<실시예 2-1> 세포의 분주≪ Example 2-1 >
세포주로서 Madin-Darby Canine kidney epithelial cell (MDCK Line)을 사용하였다. 96-well plate의 각 well에 배양액(cell growth media; MEM ＋ 10% FSB ＋ 100 U/mL penicillin ＋ 0.1 mg/mL streptomycin) 100 μL를 넣고, 세포주(MDCK cell)을 1.5 x 105 cells/ml 농도로 96-well plate에 분주한 후 세포밀도(cell confluence)가 70% - 80% 되면 사용하였다.
Madin-Darby canine kidney epithelial cell (MDCK Line) was used as a cell line. 96-well plate culture medium to each well of a (cell growth media; MEM + 10 % FSB + 100 U / mL penicillin + 0.1 mg / mL streptomycin) into the 100 μL, a cell line (MDCK cell) 1.5 x 10 5 cells / ml And the cell density (cell confluence) was 70% -80%.
<< 실시예Example 2-1> 세포독성 평가 2-1> Evaluation of cytotoxicity
세포독성을 평가하기 위해 대파 추출물의 고형물함량을 10.000 mg/mL 내지 0.0015mg/mL 사이로 하여 처리한 후, 37℃에서 48시간 동안 5% CO2 배양기로 세포주를 배양하고, 생존하는 세포주를 MTT 법으로 염색하고 흡광도를 측정하였다.
To evaluate cytotoxicity, the solids content of L. elp extract was adjusted to between 10.000 mg / mL and 0.0015 mg / mL, and then incubated at 37 ° C for 48 hours in 5% CO 2 The cell line was cultured with an incubator, and the surviving cell line was stained with MTT method and absorbance was measured.
대파 추출물의 세포독성 평가 결과는 [도 1]에서 나타내고 있다. 세포주에 대파 추출물을 첨가한 경우, 대파 추출물의 고형물함량이 희석되어 감소할수록 흡광도는 완만하게 상승하였다. 따라서 대파 추출물은 고형물 0.3704 mg/mL 이하에서는 세포독성이 낮은 것으로 나타났다.
The results of the cytotoxicity evaluation of the lobster extract are shown in [Fig. 1]. When the extract was added to the cell line, the absorbance gradually increased with decreasing solids content of the extract. Therefore, the extracts of Corynebacterium leprae showed low cytotoxicity at a concentration of 0.3704 mg / mL or less.
<실시예 2-2> 인플루엔자 바이러스 억제 효과Example 2-2 Influenza Virus Inhibitory Effect
항인플루엔자 효과를 보기 위해, [도 1]과 같이 대파 추출물의 고형물함량을 10.000 mg/mL 내지 0.0015mg/mL 사이로 하여 처리하고, H1N1(PR8) 인플루엔자 바이러스를 접종하였다. 이에 대한 대조구 실험으로 [도 2]와 같이 타미플루 (TAMIFLU, oseltamivir phosphate) 를 농도별로 처리하고 H1N1(PR8) 인플루엔자 바이러스를 접종하였다. 이들을 37℃에서 48시간 동안 5% CO2 배양기로 세포주를 배양하고, 생존하는 세포주를 MTT 법으로 염색하고 흡광도를 측정하였다.
In order to examine the anti-influenza effect, the solid content of the larvae extract was adjusted to be between 10.000 mg / mL and 0.0015 mg / mL as in [Fig. 1], and H1N1 (PR8) influenza virus was inoculated. As a control experiment, tamiflu (TAMIFLU, oseltamivir phosphate) was treated at different concentrations as shown in Fig. 2 and H1N1 (PR8) influenza virus was inoculated. The cells were cultured in a 5% CO 2 incubator at 37 ° C for 48 hours, and the surviving cell line was stained with MTT method and absorbance was measured.
대파 추출물의 항인플루엔자 효과는 [도 1]에서 나타내고 있다. 대파 추출물의 농도 10.000 mg/mL 에서는 흡광도가 낮아서 살아있는 세포주가 적은 것으로 나타났다. 반면 대파 추출물의 고형물이 희석된 농도에서는 살아있는 세포주에 의한 흡광도가 상승하였으며, 대파 추출물의 고형물 0.0046 mg/mL에서 흡광도는 최대치 0.9에 도달하여 인플루엔자 바이러스를 최대로 억제하는 효능을 보였다. 또한 [도 2]의 타미플루를 처리한 대조구 실험과 비교했을때, 타미플루 농도 0.0046 mg/mL에서 살아있는 세포주에 의한 흡광도는 1.1 를 보임으로서, 이 농도에서 대파 추출물은 타미플루 효능의 약 81% (대파 추출물의 흡광도 0.9 /타미플루의 흡광도 1.1)를 보였다.
The anti-influenza effect of the lobster extract is shown in Fig. At a concentration of 10.000 mg / mL, the absorbance was lower and the number of living cell lines was smaller. On the other hand, the absorbance of live cell line was increased at the diluted concentration of the red ginseng extract, and the maximum absorbance at the concentration of 0.0046 mg / mL of the red ginseng extract was 0.9, which showed the maximum inhibitory effect against influenza virus. Also, when compared with the control experiment in which Tamiflu treated in [Fig. 2] At a concentration of 0.0046 mg / mL, the absorbance of the live cell line was 1.1, which showed that about 81% of the Tamiflu efficacy (the absorbance of the Larval extract 0.9 / the absorbance of the Tamiflu 1.1) was greater at this concentration.
<< 실시예Example 3> 3>
대파 추출물의 항염증 효과Antiinflammatory effect of lobster extract
<<
실시예Example
3-1> 마우스 3-1> Mouse
골수유래Bone marrow origin
대식세포 분리 및 배양 Macrophage isolation and culture
마우스(C57BL6, orient bio)를 안락사 시킨 후, 대퇴골과 경골을 분리한다. 분리된 뼈를 구멍 내어 주사기로 골수를 추출한다. 추출된 골수는 RBC lysis buffer (intron biotech, Seoul, Korea) 5ml을 넣어 적혈구를 파괴시켰다. 적혈구가 제거된 골수 세포를 L929 (한국세포주은행) 배양약 30%가 첨가된 DMEM (10% FBS)에서 1주일간 대식세포로 분화시켰다. 대식세포 (106 cells)를 12-well-plate에 후 배양후, 인플라마좀 활성 실험을 수행하였다.
After euthanating the mouse (C57BL6, orient bio), the femur and tibia are separated. Separate bone is pierced and bone marrow is extracted with a syringe. RBC lysis buffer (intron biotech, Seoul, Korea) was added to the extracted bone marrow to destroy red blood cells. Bone marrow cells from which erythrocytes were removed were differentiated into macrophages for 1 week in DMEM (10% FBS) supplemented with approximately 30% of L929 (Korea Cell Line Bank) culture. Macrophages (10 6 cells) were post-cultured on a 12-well plate and inflammation activity experiments were performed.
<< 실시예Example 3-2> 3-2> 인플라마좀Inflammase 활성 평가 Activity evaluation
대표적 염증 매개 단백질인 인플라마좀 활성 억제 효과를 관찰하기 위해 인위적인 인플라마좀 (NLRP3 inflammasome) 활성제인 Adenosine triphosphate (ATP, invivogen, Sandiego, CA, USA) 2mM를 골수유래대식세포 (Bone Marrow Derived Macrophages, BMDM) 에 처리한 후, 대파 추출물에 의한 인플라마좀 활성체인 IL-1β의 세포 배출량을 측정하여 평가하였다.
In order to observe the inhibitory effect of the inflammatory mediator protein, 2mM of an artificial influenza (NLRP3 inflammatory) activator, adenosine triphosphate (ATP, invitogen, Sandiego, CA, USA) was injected into bone marrow derived macrophages BMDM), and then the amount of IL-1β produced by the infantile extract was measured and evaluated.
골수유래 대식세포에 LPS (10ug/ml, Sigma-aldrich)를 3시간 처리하여 IL-1β 전구체 (pro-IL-1β)를 생성시켰다. 그 후 RMPI (Wellgene, Seoul, Korea)에 인플라마좀 활성 약물인 ATP (2mM) 및 대파 추출물을 동시에 1시간 처리한 후, 그 배양액을 수집한다. 또한 세포내 단백질을 추출하였다.
IL-1β precursor (pro-IL-1β) was produced by treating LPS (10 ug / ml, Sigma-aldrich) for 3 hours. Thereafter, RMPI (Wellgene, Seoul, Korea) is treated with the active drug, ATP (2 mM) and lupine extract, for 1 hour at the same time, and the culture solution is collected. In addition, intracellular proteins were extracted.
수집된 배양액과 세포내 단백질을 PAGE-gel (16% or 10%)에서 전기 영동하였다. 그 후, PVDF membrane (Pall Corporation, USA)로 단백질을 옮긴다. Membrane을 3% 탈지분유에 항체가 (IL-1β [R&D system] 또는 Actin [Santa cruz biotech], 1:1000)들어 있는 용액에서 하루 저녁 교반 배양하였다. 다음 날, PBS를 이용하여 세척한 후, HRP가 붙어 있는 이차항체 (Santa cruz biotech, 1: 10,000) 용액에서 3시간 배양하였다. 반응이 끝난 후, membrane은 PBS를 이용하여 세척하고 ECL solution (Philekore)과 반응시켜 발광분석장기 (Ez-Capture, Atto)를 이용하여 분석하였다.
The collected cultures and intracellular proteins were electrophoresed on PAGE-gel (16% or 10%). The protein is then transferred to a PVDF membrane (Pall Corporation, USA). Membranes were incubated with 3% skimmed milk powder overnight in the solution containing antibody (IL-1β [R & D system] or Actin [Santa Cruz biotech], 1: 1000) overnight. The next day, the cells were washed with PBS and cultured for 3 hours in a secondary antibody (Santa Cruz biotech, 1: 10,000) with HRP. After completion of the reaction, the membrane was washed with PBS and reacted with an ECL solution (Philekore), and analyzed using a luminescence assay (Ez-Capture, Atto).
그 결과, 대파 추출물은 자체적인 인플라마좀 활성이 관찰되지 않았으며, 실험에 사용된 대파 추출물의 전체 농도에서는 대식세포 독성이 관찰되지 않았다.[도 3],[도 4]및[도 5]에서 보는 바와 같이, 본 발명의 대파 추출물은 0.625mg/mL 이상 농도에서 효과적인 인플라마좀 억제 능력이 관찰되었다.
As a result, it was found that the intestinal extract did not exhibit its own in fl ammase activity, and macrophage toxicity was not observed in the total concentration of the intestinal extract used in the experiments. [Figure 3], [Figure 4] and [Figure 5] , The L. extract of the present invention showed an effective ability to inhibit the inflammase at a concentration of 0.625 mg / mL or more.
이에 따라 본 발명은 대파 추출물의 항인플루엔자 바이러스 및 항염증 효과에 대한 것으로, 더욱 상세하게는 대파 추출물을 유효 성분으로 포함하는 인플루엔자 바이러스 감염 및 염증성 질환 예방 및 치료용 약학적 조성물 및 건강 기능성 식품 조성물을 제공한다. 본 발명은 대파 추출물의 항바이러스 및 항염증 효과를 이용하여 인플루엔자 바이러스 감염 및 염증성 질환을 예방, 개선 및 치료하는데 탁월한 효과가 있어 산업상 이용 가능성이 높다.Accordingly, the present invention relates to an anti-influenza virus and anti-inflammatory effect of a Robinia pseudo-acacia var. Umbraculifera extract, and more particularly to a pharmaceutical composition and a health functional food composition for influenza virus infection and inflammatory disease prevention and treatment, to provide. INDUSTRIAL APPLICABILITY The present invention has an excellent effect for prevention, improvement and treatment of influenza virus infection and inflammatory diseases by using the antiviral and anti-inflammatory effects of the lily extract, and thus it is highly industrially applicable.
Claims (7)
The present invention relates to a method for treating osteoarthritis, gout, ankylosing spondylitis, psoriatic arthropathy, caustic gout, familial Mediterranean fever, Behcet's disease, multiple sclerosis, rheumatoid arthritis, Which is selected from the group consisting of Muckle-Wells syndrome, adult-onset Still's disease, hyperimmunoglobulin d syndrome, and cryopyrin-associated periodic syndromes. A pharmaceutical composition for the prophylaxis and treatment of one inflammatory related inflammatory disease.
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| JP2011088881A (en) * | 2009-10-26 | 2011-05-06 | Sasaki Shokuhin Kogyo Kk | Anti-influenza viral agent and food and drink or drug including the same substance |
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| KR20180032765A (en) | 2016-09-23 | 2018-04-02 | 한국식품연구원 | A composition for improving, preventing and treating of fatty liver diseases comprising leek extract |
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