KR101951010B1 - Composition for enhancing innate immunity and antivirus comprising Isatidis Folium extract as effective component - Google Patents
Composition for enhancing innate immunity and antivirus comprising Isatidis Folium extract as effective component Download PDFInfo
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- KR101951010B1 KR101951010B1 KR1020150042938A KR20150042938A KR101951010B1 KR 101951010 B1 KR101951010 B1 KR 101951010B1 KR 1020150042938 A KR1020150042938 A KR 1020150042938A KR 20150042938 A KR20150042938 A KR 20150042938A KR 101951010 B1 KR101951010 B1 KR 101951010B1
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Abstract
본 발명은 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 약학 조성물, 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 건강기능식품, 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 사료 조성물 및 대청엽 추출물을 인간을 제외한 선천면역 증진이 필요한 동물에 투여하는 것을 포함하는 동물의 선천면역 증진 및 항바이러스 활성 증진 방법에 관한 것이다. 본 발명에 따른 선천면역 증진 및 항바이러스용 대청엽 조성물은 항바이러스 활성 및 선천면역 증진 효능을 나타내어, 박테리아 및 바이러스 감염성 질환의 예방 및 치료용으로 적용될 수 있고, 면역이 저하되거나 면역이 억제된 환자의 면역력 증진 및 조절을 위한 의약이나 건강기능식품, 및 동물의 항 질병 강화를 목적으로 하는 선천면역 증진 및 항바이러스용 사료 등으로 광범위하게 이용될 수 있다. 또한, 본 발명에 따른 선천면역 증진 및 항바이러스용 조성물은 독성이나 부작용을 거의 일으키지 않으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있다.The present invention relates to a concomitant immunization enhancing and antiviral pharmaceutical composition comprising an extract of Daechung Leaf as an active ingredient, a congenital immunity enhancing and antiviral health functional food containing Daechung Leaf extract as an active ingredient, And a method for promoting innate immune enhancement and antiviral activity in an animal, which comprises administering to a mammal in need of enhancement of innate immunity other than humans, an animal feed composition and an antiviral feed composition. The composition for congenital immunity enhancement and antiviral composition according to the present invention exhibits antiviral activity and innate immune enhancing effect and can be applied for prevention and treatment of bacterial and viral infectious diseases, , A medicinal or health functional food for enhancing and controlling immunity, and a condom immunization enhancer and an antiviral feed for enhancing an anti-disease of an animal. In addition, since the composition for congenital immunity enhancement and antiviral according to the present invention hardly causes toxicity or side effects, it can be safely used for prolonged use even for prophylactic purposes.
Description
본 발명은 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스 조성물에 관한 것으로, 더욱 상세하게는 약학, 건강기능식품 또는 사료 조성물로 이용될 수 있는 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스 조성물에 관한 것이다.The present invention relates to a congenital immunity enhancing and antiviral composition containing an extract of the broadleaf leaf extract as an active ingredient, and more particularly to a connexin immunoenhancement and antiviral composition containing an extract of a broadleaf leaf which can be used as a pharmaceutical, a health functional food or a feed composition, Immunogenic and antiviral compositions.
면역은 태어날 때부터 지니고 있는 선천 면역(innate immunity)과 후천적으로 생활하면서 적응되어 얻어지는 획득 면역(acquired immunity)으로 구분될 수 있다. 선천 면역은 일명 '자연 면역'이라고도 하며, 항원에 대해 비특이적으로 반응하고, 특별한 기억작용은 없는 것을 특징으로 한다.Immunity can be divided into innate immunity, which is born from birth, and acquired immunity, which is obtained by adapting to life. Congenital immunity is also called "natural immunity" and is characterized by nonspecific responses to antigens and no special memory effect.
최근 들어, 생체 내 선천적 방어 면역시스템 중에서 인터페론(interferon)에 의해 유도되는 선천성 면역분야가 주목을 받고 있는데, 인터페론 매개 면역의 활성화는 다양한 전염병 병원체에 대한 근본적인 예방 방법이 될 수 있기 때문이다. 이에, 인터페론 활성화 기전 연구 및 인터페론을 유도시킬 수 있는 면역조절제제의 개발 연구가 활발하게 진행되고 있다. Recently, interferon-derived congenital immunity has been attracting attention in the in vivo innate defense immune system because activation of interferon-mediated immunity can be a fundamental preventive measure against various infectious pathogens. Therefore, studies on the interferon activation mechanism and development of immunoregulatory agents capable of inducing interferon have been actively conducted.
한편, 병원체의 감염에 따른 선천 면역의 방어 기작의 하나로 사이토카인 같은 면역 인자가 분비되는데, 이들에 의해 면역 반응이 일어나고 병원체에 대한 방어가 이루어진다. 따라서 선천 면역 반응을 유도함으로써 다양한 전염병 병원체 대한 예방 및 치료 방법이 될 수 있으며, 이를 유도시킬 수 있는 선천면역의 증진 제제에 대한 연구가 필요하다.On the other hand, immune factors such as cytokines are secreted as a defense mechanism against congenital immune due to infection of pathogens. Immune response occurs and defense against pathogens occurs. Therefore, it can be a preventive and therapeutic method for various infectious disease pathogens by inducing innate immune response, and it is necessary to study the enhancing agent for congenital immune which can induce it.
바이러스(virus)는 라틴어로 독성물질을 의미하며, 세균여과지(0.22㎛)를 통과하는 일군의 감염형 병원성 입자이다. 바이러스는 숙주세포의 종류에 따라 박테리오파지, 식물 바이러스, 동물 바이러스로 분류하기도 하며, 핵산의 종류에 따라 DNA 바이러스, RNA 바이러스로 분류할 수 있다. 최근 신종 플루, AI 및 구제역 등 다양한 바이러스 질병이 사회적으로 큰 문제를 일으켰으며, 이에 따라 바이러스 질병의 효과적인 대책에 대한 고민이 사회적으로 큰 관심을 불러일으키고 있다. Virus is a Latin word for toxic substances, a group of infectious pathogenic particles that pass through bacterial filter paper (0.22 μm). Viruses can be classified into bacteriophages, plant viruses, and animal viruses according to the type of host cell. DNA viruses and RNA viruses can be classified according to the type of nucleic acid. Recently, various virus diseases such as H1N1, AI, and foot-and-mouth disease have caused a great social problem, and the concern about effective measures for viral diseases has raised a great interest in society.
현재 바이러스성 질병을 예방하기 위한 가장 좋은 방법은 백신접종이지만, 바이러스에 의한 질병의 경우, 대체로 많은 바이러스 혈청형(아형) 생성 등에 따른 백신의 효율성 문제가 중요하게 제기되고 있다. 이러한 백신의 문제점을 보완해 줄 수 있는 바이러스 예방용 억제제의 개발 및 보급은 중요한 사항이며, 이를 위해 특히 바이러스에 대한 초기 방어시스템인 생체 내 선천적 면역시스템을 자극하여 개체 동물의 면역력을 높여주는 예방제제의 발굴 및 개발은 중요한 제제 개발 방법이 될 수 있다.Currently, vaccination is the best way to prevent viral diseases. However, in case of viral diseases, vaccine efficiency due to the generation of many viral serotypes (subtypes) is important. The development and dissemination of antiviral inhibitors that can overcome the problems of these vaccines is an important issue. For this purpose, a preventive agent that enhances immunity of an individual animal by stimulating the in vivo innate immune system, Can be an important method of drug development.
인플루엔자바이러스의 증식을 억제하는 대표적인 항바이러스 제제로는 아만타딘(amantadine)과 리만타딘(rimantadine)이 있으나, 이들 두 가지 항바이러스 제제들은 혈청형 A형 인플루엔자바이러스에만 효과적이며, M2 단백질이 없는 혈청형 B형 인플루엔자바이러스에는 효과가 없는 것으로 확인되었다. 또한, 아만타딘과 리만타딘은 사용 시 인플루엔자바이러스 M2 단백질의 이온채널기능에 영향을 미치지 못하는 변이 바이러스의 출현이 매우 쉽게 일어나는 단점이 있는 것으로 확인되고 있다. 이러한 단점을 보완하기 위하여 16종의 모든 혈청형 A형 인플루엔자바이러스와 혈청형 B형 인플루엔자바이러스에 효과적인 항바이러스 제제로 자나미비르(zanamivir)와 오셀타미비르(oseltamivir)가 개발되었다. 그러나 자나미비르는 흡입 및 정맥 투여해야 하는 단점이 있으며, 오셀타미비르는 경구투여가 가능하나 최근 내성 바이러스의 출현 보고와 경구투여 시 구토와 현기증 등의 부작용이 있어 단점으로 지적되고 있다.Amantadine and rimantadine are two typical antiviral agents that inhibit the proliferation of influenza virus. However, these two antiviral agents are effective only for the serotype A influenza virus and the serotype B It has been confirmed that it is not effective against influenza virus. In addition, amantadine and rimantadine have been found to have a disadvantage in that the mutant virus, which does not affect the ion channel function of the influenza virus M2 protein, appears very easily when used. To overcome this drawback, zanamivir and oseltamivir have been developed as antiviral agents effective against all 16 serotype A influenza viruses and serotype B influenza viruses. However, there is a disadvantage that Zanamivir should be administered by inhalation and intravenous injection. Ocelaminivir can be administered orally, but it is pointed out as a disadvantage due to recent reports of the emergence of resistant virus and side effects such as vomiting and dizziness when administered orally.
또한, 수포성구내염바이러스(Vesicular stomatitis virus)의 주된 통제 방법은 질병의 완전 치료가 불가능하기 때문에, 구제역과 마찬가지로 예방 및 차단 방역과 발생 지역 내 감수성 가축의 박멸이 최선의 방법이다.In addition, as the main control method of vesicular stomatitis virus is the inability to completely cure diseases, prevention and blocking prevention as well as foot-and-mouth disease and eradication of susceptible domestic livestock are the best methods.
또한, 뉴캐슬병 바이러스에 대한 백신은 크게 생독 백신과 사독 백신으로 구분되는데, 가장 널리 이용되어온 대표적인 뉴캐슬병 생독 백신주인 B1주와 La Sota주(Clone주 포함)는 백신 접종 반응을 유발하는 것으로 알려져 있으며, 전 세계적으로 사용되고 있는 뉴캐슬병 사독 백신인 다가 혼합 사독 오일 백신은 1회 백신 접종으로 3종 이상의 질병이 동시에 예방될 수 있으나, 산란계 농장의 경우 면역력 저하로 인한 뉴캐슬병 발생 피해 사례가 늘어나고 있다. In addition, vaccines against Newcastle Disease virus are classified into virulence vaccine and Sadox vaccine. It is known that the most widely used Newcastle disease virulence vaccine, B1 strain and La Sota strain (including Clone strain) The multivitamins combined vaccine oil vaccine, a Newcastle Disease vaccine that is used globally, can prevent three or more diseases at the same time by a single vaccination. However, in the case of laying hens farms, cases of Newcastle disease caused by immunity reduction are increasing.
또한, RNA 바이러스 가운데에서도 비교적 크기가 작은 바이러스들이 있다. 이들은 작다는 뜻의 'pico'라는 말과 'RNA'를 합쳐 피코나바이러스라고 부르며, 여기에 속한 바이러스들을 통틀어 피코나바이러스과 (Picona viridae)라고 부른다. 피코나바이러스 과에 속하는 엔테로바이러스는 무균성 수막염, 수족구병, 포진성 구협염, 확장성 심근염, 급성 출혈성 결막염 등의 다양한 임상증상을 일으키는 약 70가지 혈청형을 포함하고 있으며, 폴리오바이러스(Poliovirus), 콕사키바이러스(Cossackie virus) 및 에코바이러스(Echo virus)와 기타 엔테로바이러스로 분류된다. 엔테로바이러스의 직경이 20~30nm 정도이며, 단일 가닥의 RNA를 유전자로 가지고 있다. 대부분이 등뼈동물의 소화기관을 비롯하여 호흡기관 및 중추신경계까지 감염되지만 뚜렷한 증상을 나타내지 않는 경우가 많다. 콕사키바이러스(Coxsackie virus, CXV)는 피코나바이러스 과에 속하는 인간 엔테로바이러스(human enterovirus)로서, 크게 A형과 B형으로 구분된다(Pallansch MA and Roos RP, Fields Virology, 4th edi, pp723-775, 2001). There are also relatively small viruses among RNA viruses. These are called 'pico', which means small, and 'RNA', which are called picornaviruses. These viruses are called picona viridae. Enteroviruses belonging to the family of picornaviruses contain about 70 serotypes that cause various clinical symptoms such as aseptic meningitis, hand-foot disease, herpetic myelitis, acute hemorrhagic conjunctivitis and poliovirus. , Cossackie virus and echo virus, and other enteroviruses. The diameter of the enterovirus is about 20 to 30 nm, and it has a single strand RNA as a gene. Most are infected to the respiratory organs and central nervous system as well as the digestive organs of the vertebrates, but often do not show any obvious symptoms. Coxsackie virus (CXV) is a human enterovirus belonging to the picornaviridae, and is largely divided into A type and B type (Pallansch MA and Roos RP, Fields Virology, 4th edi, pp723-775 , 2001).
또한, 최근 세계 곳곳에서 고 위험성 엔테로바이러스 및 변종 바이러스(엔테로바이러스 71형(EV-71), 콕사키바이러스 A24 변이주)가 새롭게 발견되어 유행되고 있어 이를 조기에 탐지하기 위한 국가 간 공동감시체계 구축이 요구되고 있다. In recent years, high-risk enteroviruses and mutant viruses (enterovirus type 71 (EV-71) and coxsackievirus A24 mutant strains) have been newly discovered and become popular all over the world. Is required.
콕사키바이러스를 포함하는 엔테로바이러스는 척추동물의 소화기관을 비롯하여 호흡기관 및 중추신경계까지 감염되어 다양한 임상증상을 유발하기 때문에 국가 차원의 대책 마련이 시급히 요구되고 있으나 바이러스의 종류와 혈청형이 매우 다양하여 효과적인 상용화 백신이나 치료제가 개발되어 있지 못한 실정이다.Enteroviruses including Coxsackie virus are infected to respiratory organs and central nervous system including vertebrate animal digestive organs and cause various clinical symptoms. Therefore, it is urgently required to prepare countermeasures at the national level. However, the types and serotypes of viruses are very various Thus, effective commercialized vaccines and therapeutic agents have not been developed.
또한, 단순포진바이러스(Herpes Simplex Virus; 이하 HSV)는 헤르페스 바이러스 속에 속하는 DNA 바이러스로서 약 175nm의 크기를 가지는 비교적 큰 바이러스이며, 인간에게 널리 퍼져 있는 감염체이다. 단순포진 감염은 HSV 1형(이하, 'HSV-1'라 함) 및 HSV 2형(이하, 'HSV-2'라 함)의 감염으로써, 점막이나 피부를 침범하는 급성 수포성 질환이며 아토피가 있는 사람에게서 흔히 볼 수 있는 감염질환이다. HSV-1은 주로 입과 안구 주위에, HSV-2는 성기주위에 수포를 형성시킨다. 이러한 HSV의 감염은 면역기능이 저하되어 있는 환자들에게는 그 정도가 심하게 진행되며 경부암의 원인으로도 작용하는 것으로 보고되어 있다(Melnick, J. L., Adam, E. and Rawls, W., Concer, 34, 1355-1385, 1974). 또한, 신생아나 태아의 경우에는 스스로 HSV 항체를 생성하지 못할 뿐 아니라 모체의 항체가 태아에게 넘어가지 못하기 때문에, 일단 감염되면 대부분 치명적인 결과를 초래하게 된다고 알려져 있다. 현재, HSV-1에 의한 안구질환의 경우는 미국 내에서만 1년에 약 50만 명의 환자가 발생하며 이중 1000명 이상은 안구 이식수술을 받고 있는 것으로 알려져 있다. HSV-2의 경우 약 95%는 활동성 병변이 있는 상대방과의 성적 접촉을 통해 감염되는 것으로 알려져 있는데, 미국 성인 중 약 20~30%가 HSV-2에 감염되어 있으며, 이중 20 내지 50%는 재발성 성기 포진을 갖는 것으로 보고되어 있다(Johnson R. E. et al., N. Engl. J. Med., 321, 7,1989). Herpes Simplex Virus (hereinafter referred to as HSV) is a relatively large virus having a size of about 175 nm as a DNA virus belonging to the genus Herpesvirus, and is an infectious agent widely spread to humans. Herpes simplex infection is an infectious disease of HSV type 1 (hereinafter referred to as 'HSV-1') and HSV type 2 (hereinafter referred to as 'HSV-2'), It is a common infectious disease in people with HSV-1 mainly forms around the mouth and eye, and HSV-2 forms blisters around the penis. In addition, it has been reported that HSV infection is a serious cause of cervical cancer in immunocompromised patients (Melnick, JL, Adam, E. and Rawls, W., Concer, 1355-1385, 1974). In addition, it is known that newborns and fetuses can not produce HSV antibodies themselves, and that antibodies to maternal antibodies can not be passed on to the fetus. Currently, about 500,000 cases of HSV-1-associated ocular disease occur in the United States alone in a year, of which more than 1,000 are known to undergo eye transplant surgery. Approximately 95% of HSV-2 infections are known to be transmitted through sexual contact with opponents with active lesions. About 20% to 30% of US adults are infected with HSV-2, of which 20 to 50% (Johnson RE et al., N. Engl. J. Med., 321, 7, 1989).
HSV는 초기 감염 후에 피부나 점막 표피세포에서 복제를 시작하여 신경조직을 따라 이동한 후, 전 생애에 걸쳐 척추 신경절에 잠복하고 있다가 면역기능이 저하될 경우 재활성화되어 감염부위로부터 여러 가지 질환을 발생시키는 것으로 알려져 있다. HSV에 의한 감염질환에 대한 치료제로는 뉴클레오사이드(nucleoside) 유도체인 아시클로비르(Acyclovir)가 초기감염에 매우 효과적인 것으로 알려져 있으나(Bryson, Y. J. et al., N. Engl. J. Med., 308, 916, 1983), 이 약제의 효과를 유지하기 위해서는 약제를 계속해서 투여해야 하며, 투여가 중단될 경우 HSV에 의한 감염질환이 재발하는 것으로 알려져 있다(Mindel A. et al., Lancet i: 926-928(1988); Straus S.E. et al., N. Engl. J. Med., 310, 1545(1984)). 또한 HSV의 최초 감염에 대한 치료 후 재발하는 경우에는 치사율이 높다고 보고되어 있다(Nahmias, A. J. and Coleman, R. M., Immunobiology of Herpes Simplex Virus Infection CRC Press, Boca Raton, 92-102,1984).After HSV infection, HSV begins to replicate in skin or mucosal epidermal cells and travels along the nervous system. After HSV infection, HSV is latent in the spinal ganglia throughout the entire life span, and reactivates when the immune function deteriorates. ≪ / RTI > Acyclovir, a nucleoside derivative, is known to be highly effective for early infections (Bryson, YJ et al., N. Engl. J. Med. 308, 916, 1983). In order to maintain the efficacy of this drug, it is necessary to continuously administer the drug, and when the administration is discontinued, the infectious disease caused by HSV recurs (Mindel A. et al., Lancet i: Straus SE et al., N. Engl. J. Med., 310, 1545 (1984)). It has also been reported that recurrence after treatment for the first infection of HSV has a high mortality (Nahmias, A.J. and Coleman, R.M., Immunobiology of Herpes Simplex Virus Infection CRC Press, Boca Raton, 92-102,1984).
지금까지 이러한 HSV의 감염을 예방하기 위한 백신이 개발되어 왔으나, 바이러스 자체를 약화시켜 사용하는 생백신(live attenuated vaccine)은 HSV의 게놈이 종양발생(oncogenesis)에 직접 관여한다는 사실이 밝혀짐으로 인해서 그 사용이 금지되어 왔다(Cappel, R., Sprecher, S., De Cuyper, F. and De Braekeleer, J., J. Med. Virol., 16, 137-145,1985).Although a vaccine has been developed to prevent the infection of HSV, live attenuated vaccine, which weakens the virus itself, has been shown to be directly involved in the oncogenesis of the HSV genome. (Cappel, R., Sprecher, S., De Cuyper, F. and De Braekeleer, J., J. Med. Virol., 16, 137-145, 1985).
이러한 상황하에서, 최근 해외 및 국내에서 기존의 항바이러스 제제의 단점을 극복하고자 하는 많은 노력이 이루어지고 있으며, 그 중의 하나로 국내에서는 생약 추출물 및 식물 추출물을 대상으로 항바이러스 효능에 대한 연구가 진행중이기는 하지만, 아직까지는 미비한 실정이므로, 기존의 항바이러스 제제의 단점을 극복하여 독성 및 부작용이 거의 없이 우수한 선천면역 증진 효과 및 항바이러스 활성을 발휘할 수 있는 생약 추출물을 유효성분으로 하는 조성물의 개발이 필요한 실정이다.Under these circumstances, much effort has recently been made to overcome the disadvantages of existing antiviral agents both at home and abroad. One of them is research on antiviral efficacy against herbal medicine extracts and plant extracts in Korea , It is necessary to develop a composition containing the herbal medicine extract as an active ingredient which can overcome the disadvantages of the existing antiviral agent and exhibit the innate immune enhancing effect and the antiviral activity with little toxicity and side effects .
한편, 대청엽(Isatidis Folium)은 십자화과에 속한 이년생 초본인 송남의 잎(엽)으로, 전초는 숭람, 뿌리는 판람근이라 하며, 청열, 해독, 양혈소반, 유행성감기, 간염, 하리, 급성폐렴, 토혈, 구갈, 황달, 이질 등에 효능이 있는 것으로 알려져 있으며(강병화, 한국생약자원생태도감, 2008, 379쪽), 한국등록특허 제1191724호에 대청엽 추출물을 함유하는 미백 및 주름개선용 조성물에 관해 개시되어 있다. 하지만 아직까지는 대청엽 추출물을 이용한 선천면역 증진 효과 및 항바이러스 활성에 관해 보고된 바 없다. Isatidis Folium is a leaf of Cygnus, a perennial herb that belongs to the cruciferous family. The outbreak is called shirring, and the roots of it are called shiram root. (Kang, Byung-Hwa, Korean Journal of Pharmacopoeia, Ecology, 2008, p. 379), Korean Patent No. 1191724 discloses a composition for improving whitening and wrinkles, . However, there has been no report on the effect of the extract of Daechung Leaf on the enhancement of innate immunity and antiviral activity.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 대청엽 추출물이 선천 면역의 주요 세포인 대식 세포를 활성화시켜 다양한 바이러스의 증식을 억제할 수 있음을 확인함으로써 본 발명을 완성하였다. 또한, 본 발명은 독성 및 부작용이 거의 없어 장기간 안전성을 확보하면서, 각종 바이러스 또는 세균에 의한 감염성 질환의 예방 또는 치료용 약학 조성물, 건강기능식품 또는 가축 사료 조성물 등에 효과적으로 이용될 수 있는 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스 조성물 및 동물의 선천면역 증진 및 항바이러스 활성 증진 방법을 제공하는 것을 그 목적으로 한다.SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and it is an object of the present invention to accomplish the present invention by confirming that the extract of Daicuspora japonica can inhibit the proliferation of various viruses by activating macrophages which are the main cells of innate immunity. In addition, the present invention relates to a composition for preventing or treating infectious diseases caused by various viruses or germs, which can be effectively used for health functional foods or animal feed compositions, It is an object of the present invention to provide a congenital immunity enhancement and antiviral composition containing as an active ingredient, a method for enhancing innate immunity and an antiviral activity of an animal.
상기 목적을 달성하기 위하여, 본 발명은 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 약학 조성물을 제공한다.In order to achieve the above-mentioned object, the present invention provides a concomitant immunity enhancing and antiviral pharmaceutical composition comprising an extract of Daicuspora Leaf as an active ingredient.
또한, 본 발명은 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 건강기능식품을 제공한다.In addition, the present invention provides a congenital immunity-promoting and antiviral health functional food containing an extract of Daechungchon leaf as an active ingredient.
또한, 본 발명은 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 사료 조성물을 제공한다.In addition, the present invention provides a congenital immunity enhancing and antiviral feed composition containing an extract of Daechungpoek as an active ingredient.
또한, 본 발명은 인간을 제외한 선천면역 증진이 필요한 동물에 대청엽 추출물을 투여하는 것을 특징으로 하는 동물의 선천면역 증진 및 항바이러스 활성의 증진 방법을 제공한다. In addition, the present invention provides a method for enhancing innate immunity and antiviral activity of an animal, which comprises administering an extract of Miscanthidae to an animal requiring convalescent immunity enhancement except for a human.
본 발명에 따르면 우수한 선천면역 증진 효능을 발휘하는 대청엽 추출물을 이용함으로써 각종 바이러스 및 세균 감염성 질병의 예방 및 치료에 효과적으로 적용될 수 있으며, 면역력 증진에 기여할 수 있다.According to the present invention, it is possible to effectively apply to the prevention and treatment of various viruses and bacterial infectious diseases by using the extract of Daechung leaf which exerts excellent innate immune enhancing effect, and can contribute to enhancement of immunity.
또한, 본 발명에 따른 대청엽 추출물을 포함하는 선천면역 증진 및 항바이러스용 조성물은 독성이나 부작용을 거의 일으키지 않으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있다. 따라서 본 발명에 따른 조성물은 박테리아 및 바이러스에 의한 감염성 질환의 예방 및 치료용으로 적용될 수 있고, 면역이 저하되거나 면역이 억제된 환자의 면역력 증진 및 조절을 위한 약학 조성물이나 건강기능식품, 및 동물의 항 질병 강화를 목적으로 하는 면역증진용 사료 조성물 등으로 광범위하게 이용될 수 있다.In addition, the composition for congenital immunity enhancement and antiviral composition containing the extract of Miscanthus multiflora according to the present invention scarcely causes toxicity or side effects, so that it can be safely used for prophylactic purposes even for a long period of time. Accordingly, the composition according to the present invention can be applied for the prevention and treatment of infectious diseases caused by bacteria and viruses, and can be applied to pharmaceutical compositions and health functional foods for immunity enhancement and control of immunocompromised patients, And an immunity enhancing feed composition aimed at enhancing anti-disease.
또한, 본 발명에 따르면 대청엽 추출물을 선천면역 증진에 필요한 동물에 투여함으로써 동물의 면역을 효과적으로 안전하게 증진시킬 수 있다.In addition, according to the present invention, it is possible to effectively and safely enhance the immunity of an animal by administering the extract of Daicuspora japonica to an animal necessary for congenital immunity enhancement.
도 1은 본 발명의 일 실시예에 따른 대청엽 추출물의 인플루엔자바이러스(PR8-GFP virus)에 대한 항바이러스 활성분석 결과이다. Medium은 음성 대조군으로 아무것도 처리하지 않은 세포군; PR8-GFP는 바이러스 감염군; IFN-β/PR8-GFP는 양성대조군으로, PR8-GFP 바이러스 감염 및 1,000Units/㎖의 IFN-β 처리군; 대청엽/PR8-GFP는 PR8-GFP 바이러스 감염 및 대청엽 추출물 처리군이다.
도 2는 본 발명의 일 실시예에 따른 대청엽 추출물의 수포성구내염바이러스(VSV-GFP virus)에 대한 항바이러스 활성분석 결과이다. Medium은 음성 대조군으로 아무것도 처리하지 않은 세포군; VSV-GFP는 바이러스 감염군; IFN-β/VSV-GFP는 양성대조군으로, VSV-GFP 바이러스 감염 및 1,000Units/㎖의 IFN-β 처리군; 대청엽/VSV-GFP는 VSV-GFP 바이러스 감염 및 대청엽 추출물 처리군이다.
도 3은 본 발명의 일 실시예에 따른 대청엽 추출물의 단순포진바이러스(HSV-GFP virus)에 대한 항바이러스 활성분석 결과이다. Medium은 음성 대조군으로 아무것도 처리하지 않은 세포군; HSV-GFP는 바이러스 감염군; IFN-β/HSV-GFP는 양성대조군으로, HSV-GFP 바이러스 감염 및 1,000Units/㎖의 IFN-β 처리군; 대청엽/HSV-GFP는 HSV-GFP 바이러스 감염 및 대청엽 추출물 처리군이다.
도 4는 본 발명의 일 실시예에 따른 대청엽 추출물의 뉴캐슬병바이러스(NDV-GFP virus)에 대한 항바이러스 활성분석 결과이다. Medium은 음성 대조군으로 아무것도 처리하지 않은 세포군; NDV-GFP는 바이러스 감염군; IFN-β/NDV-GFP는 양성대조군으로, NDV-GFP 바이러스 감염 및 1,000Units/㎖의 IFN-β 처리군; 대청엽/NDV-GFP는 NDV-GFP 바이러스 감염 및 대청엽 추출물 처리군이다.
도 5는 본 발명의 일 실시예에 따른 대청엽 추출물의 엔테로바이러스(EV-71 virus)에 대한 항바이러스 활성분석 결과이다. Medium은 음성 대조군으로 아무것도 처리하지 않은 세포군; EV-71는 바이러스 감염군; IFN-β/EV-71은 양성대조군으로, EV-71 바이러스 감염 및 1,000Units/㎖의 IFN-β 처리군; 대청엽/EV-71는 EV-71 바이러스 감염 및 대청엽 추출물 처리군이다.
도 6은 본 발명의 실시예에 따른 대청엽 추출물에 의한 전 염증성 사이토카인 유도 분석결과를 나타낸다.
도 7은 본 발명의 실시예에 따른 대청엽 추출물에 의한 인플루엔자바이러스(H1N1 및 H5N2)의 감염 억제를 확인한 결과이다. H1N1 및 H5N2는 인플루엔자바이러스만 처리(1.0 MOI)된 군이고, Medium은 음성대조군으로 아무것도 처리되지 않은 MDCK 세포군이며, 대청엽/H1N1 및 대청엽/H5N2는 1㎍/㎖의 대청엽 추출물과 인플루엔자바이러스(1.0 MOI)를 동시에 처리한 군이다.FIG. 1 shows the results of analysis of antiviral activity against influenza virus (PR8-GFP virus) of the extract of Daicuspora leaf according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; PR8-GFP is a group of virus infections; IFN-? / PR8-GFP as a positive control, PR8-GFP virus infection and 1,000 units / ml IFN-? Treatment group; Daechung Leaf / PR8-GFP is a group treated with PR8-GFP virus infection and extract of Daechung leaf.
FIG. 2 is a graph showing the results of antiviral activity assay of VSV-GFP virus of the extract of Daicuspora japonica extract according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; VSV-GFP is a virus-infected group; IFN-β / VSV-GFP was a positive control group, with VSV-GFP virus infection and 1,000 units / ml IFN-β treatment group; Daechungpo leaf / VSV-GFP is a group treated with VSV-GFP virus infection and extracts of Daechung leaf.
FIG. 3 shows the results of analysis of antiviral activity against herpes zoster virus (HSV-GFP virus) of the extract of Daicuspora leaf according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; HSV-GFP is a virus-infected group; IFN-β / HSV-GFP as a positive control, HSV-GFP virus infection and 1,000 units / ml IFN-β treatment group; HSC-GFP is a group treated with HSV-GFP viral infection and extracts of macrochannel leaves.
FIG. 4 shows the results of analysis of antiviral activity against Newcastle Disease Virus (NDV-GFP virus) according to one embodiment of the present invention. Medium is a negative control group of cells that have not been treated; NDV-GFP is a virus-infected group; IFN-? / NDV-GFP is a positive control group, NDV-GFP virus infection and 1,000 units / ml IFN-? Treatment group; Daechung leaf / NDV-GFP is a group treated with NDV-GFP virus infection and extracts of Daechung leaf.
FIG. 5 is a graph showing the results of analysis of antiviral activity against the enterovirus (EV-71 virus) of the extract of Daechungchalafloff according to an embodiment of the present invention. Medium is a negative control group of cells that have not been treated; EV-71 is a virus-infected group; IFN-beta / EV-71 was a positive control group, with EV-71 viral infection and 1,000 Units / ml IFN-? Treatment group; Daechung Leaf / EV-71 is a group treated with EV-71 virus infection and Daechung leaf extract.
FIG. 6 shows the results of proinflammatory cytokine induction assay by the extract of Daechung Lob, according to an embodiment of the present invention.
FIG. 7 shows the results of confirming the inhibition of influenza virus (H1N1 and H5N2) infection by the extract of Daechung leaf according to an embodiment of the present invention. H1N1 and H5N2 were treated only with influenza virus (1.0 MOI). Medium was a negative control MDCK cell group, and Daechungpo leaf / H1N1 and Daechung leaf / H5N2 were treated with 1 ㎍ / (1.0 MOI) at the same time.
본 발명은 대청엽 추출물을 유효성분으로 함유하는 선천면역 증진 및 항바이러스용 약학 조성물에 관한 것이다. The present invention relates to a concomitant immunity enhancing and antiviral pharmaceutical composition containing an extract of Miscanthidae as an active ingredient.
상기 대청엽 추출물은 물, 탄소수 1 내지 4의 알코올 중에서 선택된 1종 이상의 용매로 추출하는 것이 바람직하고, 더 바람직하게는 물, 메탄올, 에탄올, 또는 부탄올의 용매를 이용하여 추출하는 것이며, 더욱더 바람직하게는 물을 용매로 이용하여 열수 추출한 것이다. 상기 열수 추출은 1) 대청엽 중량을 기준으로, 5 내지 30배의 증류수를 첨가하는 단계; 2) 100~130℃의 온도에서 2~5시간 동안 열수 추출하는 단계; 및 3) 상기 열수 추출물을 여과하는 단계;를 거쳐 수행하는 것이 바람직하지만 이에 한정하지 않는다.Preferably, the extract is extracted with one or more solvents selected from water and alcohols having 1 to 4 carbon atoms, more preferably extracted with a solvent of water, methanol, ethanol, or butanol, Is a hot-water extraction using water as a solvent. The hot water extraction is performed by 1) adding 5 to 30 times of distilled water on the basis of the large leaf weight; 2) hot water extraction at a temperature of 100 to 130 ° C for 2 to 5 hours; And 3) filtering the hot-water extract. However, the present invention is not limited thereto.
또한, 상기 대청엽 추출물은 추출처리에 의해 얻어지는 추출액, 추출액의 희석액 또는 농축액, 추출액을 건조하여 얻어지는 건조물, 또는 조정제물 또는 정제물 중 어느 하나를 포함하는 것으로 한다.In addition, the above-mentioned extracts of Daicusporae leaf include any of the extract obtained by the extraction treatment, the diluted or concentrated liquid of the extract, the dried product obtained by drying the extract, or the adjusted product or the purified product.
본 발명의 대청엽 추출물이 항바이러스 활성을 갖는 바이러스의 일례는 오르토믹소비리대(Orthomixoviridae), 랍도비리대(Rhabdoviridae), 파라믹소비리대(Paramixoviridae), 허피스비리대(Herpesviridae) 및 피코나비리대(Picornaviridae) 중에서 선택된 하나 이상의 바이러스이며, 바람직하게는 인플루엔자바이러스, 뉴캐슬병바이러스, 수포성구내염바이러스(Vesicular stomatitis virus), 콕사키바이러스(Cossackie virus), 엔테로바이러스 71형(Enterovirus-71), 단순포진바이러스(Herpes Simplex Virus), 리노바이러스(Rhinovirus), 호흡기세포융합바이러스(respiratory syncytial virus; RSV), 구제역바이러스(Foot and mouth disease virus), 콜로라도참진드기열바이러스, 레오바이러스, 인간면역 결핍 바이러스, B형 간염바이러스, C형 간염바이러스, 돼지열병바이러스, 소바이러스성설사바이러스(Bovine Viral Diarrhea Virus), 돼지생식기호흡기증후군바이러스(Porcine reproductive and respiratory syndrome virus), 돼지오제스키병바이러스, 로타바이러스, 파보바이러스, 돼지유행성설사바이러스(Porcine epidemic diarrhea virus) 등이 있으며, 더 바람직한 바이러스의 일례는 인플루엔자바이러스, 뉴캐슬병바이러스, 수포성구내염바이러스, 단순포진바이러스, 엔테로바이러스 71형, 리노바이러스(Rhinovirus), 호흡기세포융합바이러스(respiratory syncytial virus; RSV)이지만 이에 제한하지 않는다.Examples of the virus having the antiviral activity of the extract of the present invention include Orthomixoviridae, Rhabdoviridae, Paramixoviridae, Herpesviridae, (Vesicular stomatitis virus, Cossackie virus, Enterovirus-71, Enterococcus faecalis, Enterococcus faecalis, Enterovirus-71, Viruses such as Herpes Simplex Virus, Rhinovirus, respiratory syncytial virus (RSV), foot and mouth disease virus, Colorado tick fever virus, reovirus, human immunodeficiency virus, B Hepatitis virus, hepatitis C virus, swine fever virus, bovine viral diarrhea virus, Porcine reproductive and respiratory syndrome virus, porcine Ozeki disease virus, rotavirus, parvovirus and porcine epidemic diarrhea virus. Examples of the more preferable virus include influenza virus, Newcastle disease virus , Vesicular stomatitis virus, herpes simplex virus,
본 발명의 대청엽 추출물은 면역 인자 유도능을 강하게 나타내며, 개체의 선천면역을 증가시켜 바이러스의 감염 및 증식을 억제하는 효과가 있을 뿐만 아니라, 세포 독성이나 부작용을 거의 나타내지 않으므로 장기간 복용시에도 안심하고 사용할 수 있는 것을 특징으로 한다. The extract of the present invention shows a strong immunity-inducing ability and increases the innate immunity of the individual to inhibit the infection and proliferation of the virus. Moreover, since it shows almost no cytotoxicity or side effects, Can be used.
본 발명의 선천면역 증진 및 항바이러스용 약학 조성물은 약학적으로 허용가능한 담체, 부형제 또는 희석제를 더 포함할 수 있다. 본 발명에 이용될 수 있는 약학적으로 허용가능한 담체, 부형제 또는 희석제는 본 발명의 효과를 해하지 않는 한 특별히 제한되지 않으며, 예를 들어 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제, 윤활제, 감미제, 방향제, 보존제 등을 포함할 수 있다. 약학적으로 허용 가능한 담체, 부형제 또는 희석제의 대표적인 예로는, 락토즈, 덱스트로스, 슈크로스, 솔비톨, 만니톨, 자일리톨, 말티톨, 전분, 젤라틴, 글리세린, 아카시아 고무, 알지네이트, 칼슘포스페이트, 칼슘카보네이트, 칼슘실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트, 광물유, 프로필렌글리콜, 폴리에틸렌글리콜, 식물성 오일, 주사가능한 에스테르, 위텝솔, 마크로골, 트윈 61, 카카오지, 라우리지 등을 들 수 있다. 본 발명의 선천면역 증진 및 항바이러스용 약학 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁액, 에멀젼, 시럽제, 에어로졸, 외용제, 좌제 및 주사제로 이루어진 군으로부터 선택되는 형태일 수 있다. 약학 조성물의 제제화 방법은 기술분야에 공지된 통상의 방법에 따라 수행될 수 있으며, 특별히 제한되지 않는다. The pharmaceutical composition for congenital immunity enhancement and antiviral of the present invention may further comprise a pharmaceutically acceptable carrier, excipient or diluent. The pharmaceutically acceptable carrier, excipient or diluent which can be used in the present invention is not particularly limited so long as the effect of the present invention is not impaired. For example, a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, Flavoring agents, fragrances, preservatives, and the like. Representative examples of pharmaceutically acceptable carriers, excipients or diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, maltitol, starch, gelatin, glycerin, acacia rubber, alginate, calcium phosphate, calcium carbonate, calcium Methylcellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, propylene glycol, polyethylene glycol, vegetable oil, injectable Ester, witepsol, macrogol, tween 61, cacao paper, and laurie paper. The pharmaceutical composition for congenital immunity enhancement and antiviral of the present invention may be in the form of a tablet, a pill, a powder, a granule, a capsule, a suspension, an emulsion, a syrup, an aerosol, a external preparation, a suppository and an injection. The method of preparing the pharmaceutical composition may be performed according to a conventional method known in the art, and is not particularly limited.
본 발명의 선천면역 증진 및 항바이러스용 약학 조성물은 경구 또는 비경구 투여될 수 있으며, 투여량은 투여 대상의 연령, 성별, 체중, 상태, 질병의 정도, 약물의 형태, 투여 경로 및 기간에 따라 적절히 선택될 수 있으나, 일반적으로 약 5~500㎎/㎏, 바람직하게는 약 100~250㎎/㎏을 1일 1~3회 투여할 수 있다.The pharmaceutical composition for congenital immunity enhancement and antiviral use according to the present invention may be administered orally or parenterally. The dosage may be adjusted according to the age, sex, weight, condition, degree of disease, drug form, May be appropriately selected, but generally about 5 to 500 mg / kg, preferably about 100 to 250 mg / kg, can be administered one to three times a day.
본 발명의 선천면역 증진 및 항바이러스용 약학 조성물의 제제화 방법, 투여량, 투여 경로, 구성성분 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다. It will be apparent to those skilled in the art that the method of concomitant immunization enhancement and formulation of pharmaceutical compositions for antiviral use of the present invention, dosage, route of administration, constituents, etc., can be appropriately selected from conventional techniques known in the art.
본 발명의 선천면역 증진 및 항바이러스용 약학 조성물은 박테리아 감염성 질병 또는 바이러스 감염성 질병의 예방 및 치료에 이용될 수 있다. 본 발명의 선천면역 증진 및 항바이러스용 약학 조성물은 유효 성분으로 대청엽 추출물 외에 다른 약학적 활성 성분을 함께 포함하거나, 또는 다른 유효 성분을 포함하는 약학 조성물과 혼합되어 이용될 수 있다. The congenital immunity enhancing and antiviral pharmaceutical compositions of the present invention can be used for the prevention and treatment of bacterial infectious diseases or viral infectious diseases. The pharmaceutical composition for congenital immunity enhancement and antiviral use of the present invention may be used as an active ingredient in combination with a pharmaceutical composition containing other active ingredients besides the extract of Miscanthidae or other active ingredients.
또한, 본 발명은 대청엽 추출물을 포함하는 선천면역 증진 및 항바이러스용 건강기능식품에 관한 것이다. 본 발명의 선천면역 증진 및 항바이러스용 건강기능식품은 식품학적으로 허용가능한 식품 보조 첨가제를 더 포함할 수 있다. 본 발명에 이용될 수 있는 식품학적으로 허용가능한 식품 보조 첨가제는 포도당, 과당, 말토오스, 슈크로스, 덱스트린, 시클로덱스트린과 같은 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당 알코올과 같은 천연 탄수화물, 타우마틴, 스테비아 추출물 등의 천연 향미제, 사카린, 아스파르탐산 등의 합성 향미제, 착색제, 펙트산 또는 그의 염, 알긴산 또는 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산화제 등을 포함하나, 이에 제한되는 것은 아니다. 본 발명의 선천면역 증진 및 항바이러스용 건강기능식품은 분말, 과립, 정제, 캡슐, 캔디, 츄잉검, 젤리 및 음료로 이루어진 군으로부터 선택되는 형태일 수 있다. 선천면역 증진 및 항바이러스용 건강기능식품 중의 대청엽 추출물의 함량은 식품의 형태, 풍미, 맛 등을 고려하여 적절하게 선택될 수 있으며, 예를 들어 건강기능식품 전체 중량에 대하여 0.01~30 중량%의 범위일 수 있다. 본 발명의 선천면역 증진 및 항바이러스용 건강기능식품의 형태, 조성 및 제조방법 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다.In addition, the present invention relates to a healthy functional food for congenital immunity enhancement and antiviral treatment including an extract of Miscanthidae. The congenital immunity enhancing and antiviral health functional food of the present invention may further include a food acceptable food supplementary additive. Food-acceptable food supplementary additives that may be used in the present invention include sugars such as glucose, fructose, maltose, sucrose, dextrin, cyclodextrins, natural carbohydrates such as sugar alcohols such as xylitol, sorbitol and erythritol, , A natural flavor such as stevia extract, a synthetic flavor such as saccharin and aspartame, a coloring agent, a pectic acid or a salt thereof, an alginic acid or a salt thereof, an organic acid, a protective colloid thickener, a pH adjusting agent, a stabilizer, Alcohols, carbonates, and the like. The congenital immunity enhancing and antiviral health functional food of the present invention may be in a form selected from the group consisting of powder, granule, tablet, capsule, candy, chewing gum, jelly and beverage. The content of the extract of Miscanthus sinensis L. in the health functional foods for congenital immunity enhancement and antiviral treatment can be appropriately selected in consideration of the shape, flavor, taste, etc. of the food, and for example, 0.01 to 30 wt% Lt; / RTI > It is apparent to those of ordinary skill in the art that the form, composition and manufacturing method of the health functional food for congenital immunity enhancement and antiviral of the present invention can be appropriately selected from conventional techniques known in the art.
또한, 본 발명은 대청엽 추출물을 포함하는 선천면역 증진 및 항바이러스용 사료 조성물에 관한 것이다. 선천면역 증진 및 항바이러스용 사료 조성물 중의 대청엽 추출물의 함량은 급여 가축의 종, 주령, 체중, 및 사육 조건 등에 따라 적절히 선택될 수 있으며, 사료 조성물 전체 중량에 대하여 0.01~95중량%, 바람직하게는 0.1~80중량%의 비율일 수 있다. 본 발명의 선천면역 증진 및 항바이러스용 사료 조성물은 기술분야에 공지된 사료 제조방법에 따라 제조될 수 있으며, 예를 들어, 각종 사료 원료 또는 배합사료와 본 발명의 대청엽 추출물을 혼합한 후, 추가적인 가공 공정, 예를 들어 펠렛 형태로의 성형 또는 과립 등의 형태로의 절단 단계 등을 더 수행함으로써 제조될 수 있다. 본 발명의 선천면역 증진 및 항바이러스용 사료 조성물의 구성성분, 조성, 제조방법, 급여방법 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다. In addition, the present invention relates to an innate immunity enhancing and anti-viral feed composition comprising an extract of Daechungchon. The content of the extract of Miscarriageae L. in the feed composition for congenital immunity enhancement and antiviral treatment can be appropriately selected according to species, age, body weight, rearing condition and the like of the feed livestock, and is 0.01 to 95% by weight, May be in a proportion of 0.1 to 80% by weight. The composition for congenital immunity enhancement and antiviral composition according to the present invention can be prepared according to a method of manufacturing feeds known in the art. For example, after mixing various feed ingredients or compound feed with the extract of Daechung leaf of the present invention, For example, by further performing a further processing step, for example, a step in the form of pellets or a step in the form of granules or the like. It is apparent to those skilled in the art that the composition, composition, manufacturing method, feeding method, and the like of the congenital immunity enhancing and antiviral feed composition of the present invention can be appropriately selected from conventional techniques known in the art.
또한, 본 발명은 인간을 제외한 선천면역 증진이 필요한 동물에 대청엽 추출물을 투여하는 것을 특징으로 하는 동물의 선천면역 증진 및 항바이러스 활성의 증진 방법에 관한 것이다. 본 발명의 동물의 선천면역 증진 및 항바이러스 활성 증진에 있어서, 대청엽 추출물의 투여량, 투여경로, 투여 시기 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다.
The present invention also relates to a method for enhancing innate immune function and antiviral activity of an animal, which comprises administering an extract of Miscanthidae to an animal in need of innate immune enhancement except for a human. It should be apparent to those skilled in the art that the dose, route of administration, administration time, etc. of the extract of Daechung leaf extract may be suitably selected from conventional techniques known in the art in enhancing innate immunity and antiviral activity of the animal of the present invention Do.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.
실시예Example 1. 대청엽 추출물의 제조 1. Preparation of extracts of Daechung Leaf
대청엽 추출물은 대청엽의 중량을 기준으로, 20배의 증류수를 대청엽에 첨가하여 115℃에서 180분간 열수 추출하고, 0.45㎛로 1차 여과한 후, 0.22㎛로 2차 여과하여 침전물을 제거하였다. 이어서, pH를 7.0으로 조정하여 1.5㎖ Ep-튜브에 1㎖씩 분주하고 -20℃에서 저장하여 모든 분석에 사용하였다.
The extracts of Daechung Leaf were extracted by hot water extraction at 115 ℃ for 180 min, and then primary filtration with 0.45 ㎛ and secondary filtration with 0.22 ㎛ to remove precipitate Respectively. Subsequently, the pH was adjusted to 7.0, and 1 ml of each was dispensed into a 1.5 ml Ep-tube and stored at -20 캜 for use in all analyzes.
실시예Example 2. 대청엽 추출물의 항바이러스 활성 분석 2. Antiviral activity analysis of extracts of Daechung Leaf
인플루엔자바이러스(Influenza virus; PR8), 수포성구내염바이러스(Vesicular stomatitis virus), 단순포진바이러스(Herpes simplex virus; HSV), 뉴캐슬병바이러스(Newcastle disease virus) 및 엔테로바이러스 71형(Enterovirus-71; EV-71)에 대한 대청엽 추출물의 항바이러스 활성 분석을 수행하였다.
Influenza virus (PR8), Vesicular stomatitis virus, Herpes simplex virus (HSV), Newcastle disease virus and enterovirus type 71 (Enterovirus-71; EV-71 ) Were tested for their antiviral activity.
(1) 항바이러스 활성 분석 방법(1) Antiviral activity assay method
인플루엔자바이러스(Influenza virus; PR8), 수포성구내염바이러스(Vesicular stomatitis virus; VSV), 단순포진바이러스(HSV-GFP) 및 뉴캐슬병바이러스(Newcastle disease virus) 를 마우스 대식세포주인 Raw 264.7 세포(8×105cell/well)에 감염시켜 분석하였고, 엔테로바이러스 71형(Enterovirus-71; EV-71)은 Hela 세포에 감염시켜 분석하였다.Influenza virus (Influenza virus; PR8), vesicular stomatitis virus (Vesicular stomatitis virus; VSV), a Raw 264.7 cells (8 × 10 5 to herpes simplex virus (HSV-GFP) and Newcastle disease virus (Newcastle disease virus) mouse macrophage cell line (Enterovirus-71; EV-71) was infected with Hela cells and analyzed.
12-웰 TC 플레이트에 세포를 배양한 후, 1% FBS가 첨가된 DMEM에 상기 실시예 1에서 제조한 1㎍/㎖의 대청엽 추출물(pH 조정)을 각각 처리하였다. 음성 대조군은 1% FBS가 첨가된 DMEM만을 처리하였고, 양성 대조군(Positive control)은 마우스 IFN-β(500units/㎖)를 처리하였다. 대청엽 추출물의 처리 12시간 후, PR8-GFP(MOI:1.0), VSV-GFP(MOI:1.0), HSV-GFP(MOI:3.0), NDV-GFP(MOI:3.0) 및 EV-71(MOI:1.0)를 각각 접종하였다. 접종하고 2시간 후 접종액을 제거하고, PBS로 3회 세척하고, 12 및 24시간 후, 바이러스 감염 정도를 확인하였다.
Cells were cultured on a 12-well TC plate and treated with 1 μg / ml of the extract prepared in Example 1 (pH adjustment), respectively, in DMEM supplemented with 1% FBS. Negative controls were treated with DMEM supplemented with 1% FBS, and positive controls were treated with mouse IFN-β (500 units / ml). GFP (MOI: 1.0), HSV-GFP (MOI: 3.0), NDV-GFP (MOI: 3.0) and EV-71 : 1.0), respectively. After 2 hours of inoculation, the inoculum was removed, washed three times with PBS, and after 12 and 24 hours, the degree of virus infection was confirmed.
(2) 항바이러스 활성 분석결과(2) Antiviral activity assay results
1) 인플루엔자바이러스(1) Influenza virus ( PR8PR8 -- GFPGFP virusvirus )의 분석결과) Analysis result
인플루엔자바이러스에 대한 항바이러스 활성 분석결과를 도 1에 개시하였다. 도 1(a)는 감염 12시간 후의 GFP(green fluorescent protein) 형광이미지를 나타내고, 도 1(b)는 감염 24시간 후의 GFP(green fluorescent protein) 형광이미지를 나타낸 것이다. 대청엽 추출물로 처리한 결과, 인플루엔자바이러스의 감염률이 현저하게 떨어진 것을 확인할 수 있었다(도 1).
The results of analysis of antiviral activity against influenza virus are shown in Fig. Fig. 1 (a) shows GFP (green fluorescent protein) fluorescence image after 12 hours of infection, and Fig. 1 (b) shows GFP (green fluorescent protein) fluorescence image after 24 hours of infection. As a result, the infectious rate of influenza virus was remarkably lowered (Fig. 1).
2) 2) 수포성구내염바이러스(VSV-GFP virus)의Of vesicular stomatitis virus (VSV-GFP virus) 분석결과 Analysis
수포성구내염바이러스에 대한 항바이러스 활성 분석 결과를 도 2에 개시하였다. 도 2(a)는 감염 12시간 후의 GFP(green fluorescent protein) 형광이미지를 나타내고, 도 2(b)는 감염 24시간 후의 GFP(green fluorescent protein) 형광이미지를 나타낸 것이다. 도 2로부터 확인할 수 있는 바와 같이, 대청엽 추출물로 처리한 결과 수포성구내염바이러스의 감염률이 현저하게 떨어진 것을 확인하였다.
The results of antiviral activity assay for vesicular stomatitis virus are shown in Fig. FIG. 2 (a) shows GFP (green fluorescent protein) fluorescence image after 12 hours of infection, and FIG. 2 (b) shows GFP (green fluorescent protein) fluorescence image after 24 hours of infection. As can be seen from FIG. 2, it was confirmed that the infection rate of the vesicular stomatitis virus was remarkably decreased when treated with the extract of Daechungpo leaf.
3) 단순포진바이러스(3) Herpes simplex virus ( HSVHSV -- GFPGFP virusvirus )의 분석결과) Analysis result
단순포진바이러스에 대한 항바이러스 활성 분석결과를 도 3에 개시하였다. 도 3(a)는 감염 12시간 후의 GFP 형광이미지를 나타내고, 도 3(b)는 감염 24시간 후의 GFP 형광이미지를 나타낸 것이다. 도 3으로부터 확인할 수 있는 바와 같이, 대청엽 추출물로 처리한 결과 단순포진바이러스의 감염률이 현저하게 떨어진 것을 확인하였다.
The results of analysis of antiviral activity against herpes simplex virus are shown in Fig. Fig. 3 (a) shows GFP fluorescence image after 12 hours of infection, and Fig. 3 (b) shows GFP fluorescence image after 24 hours of infection. As can be seen from FIG. 3, it was confirmed that the infection rate of herpes simplex virus was significantly lowered by treating with the extract of Daechunggol.
4) 뉴캐슬병바이러스(NDV-GFP virus)의 분석결과 4 ) Analysis of Newcastle disease virus (NDV-GFP virus)
뉴캐슬병바이러스에 대한 항바이러스 활성 분석결과를 도 4에 개시하였다. 도 4(a)는 감염 24시간 후의 GFP 형광이미지를 나타내며, 도 4(b)는 상대적인 GFP 형광량을 나타낸 것이다. 도 4에 개시한 바와 같이, 뉴캐슬병바이러스가 감염된 세포에 대청엽 추출물을 처리한 결과, 뉴캐슬병바이러스가 현저하게 감소한 것을 확인하였다.
The results of antiviral activity assay for Newcastle disease virus are shown in FIG. Fig. 4 (a) shows GFP fluorescence image after 24 hours of infection, and Fig. 4 (b) shows the relative GFP type light amount. As shown in FIG. 4, when the cells infected with the Newcastle disease virus were treated with the extract of the great blueberry leaf, it was confirmed that the Newcastle disease virus was markedly decreased.
5) 엔테로바이러스 ( EV -71 virus ) 71형의 분석결과 5 ) Results of analysis of enterovirus ( EV- 71 virus )
엔테로바이러스 71형에 대한 항바이러스 활성 분석결과를 도 5에 개시하였다. 도 5(a)는 감염 12시간 후의 광학이미지를 나타내고, 도 5(b)는 감염 24시간 후의 광학이미지를 나타낸 것이다. 도 5에 개시한 바와 같이, 엔테로바이러스 71형이 감염된 세포에 대청엽 추출물을 처리한 결과, 엔테로바이러스 71형이 현저하게 감소한 것을 확인하였다.
The results of antiviral activity assay for
실시예Example 3. 마우스 대식 세포주를 이용한 대청엽 3. Macrophage from mouse macrophage line 의of 전 염증성 사이토카인 유도( Proinflammatory cytokine induction ( propro -- inflammatoryinflammatory CytokineCytokine InductionInduction ) 분석) analysis
대청엽 추출물의 면역인자 유도효과를 확인하기 위하여 전 염증성 사이토카인 유도 분석을 수행하였다.The proinflammatory cytokine induction assay was performed to confirm the induction of the immune factor induction effect of the extracts of Daechungchung.
(1) 전 염증성 사이토카인 유도 분석방법(1) proinflammatory cytokine induction assay method
마우스 대식세포주인 Raw 264.7을 키워 분석에 사용하였다. 6-웰 TC 플레이트에 세포를 배양한 후, 1% FBS가 첨가된 DMEM에 상기에서 제조된 대청엽 추출물(pH 조정)을 첨가하여 처리하였다. 음성 대조군은 1% FBS가 첨가된 DMEM만을 처리하였고, 양성 대조군은 바이러스, 암세포 등의 외부 물질에 반응하여 분비되는 사이토카인으로, 세포 내 항암 및 항바이러스 작용을 일으켜 면역반응을 유도하는 물질인 인터페론-β(IFN-β)를 1000Units/㎖ 처리하였다. 시료는 1㎍/㎖의 대청엽 추출물을 처리하고 12시간 후와 24시간 후의 세포에 대하여 TNF-α, IL-6 및 IFN-β의 생성량(pg/㎖)을 ELISA로 측정하였다.
Mouse macrophage line Raw 264.7 was used for the analysis. Cells were cultured on a 6-well TC plate, and then treated with the above-prepared macrophyll extract (pH adjusted) in DMEM supplemented with 1% FBS. The negative control group was treated with DMEM supplemented with 1% FBS, and the positive control group was a cytokine secreted in response to external substances such as viruses and cancer cells, and the interferon, which is an immune response inducing substance, -β (IFN-β) was treated at 1000 units / ml. The amount of TNF-α, IL-6 and IFN-β produced (pg / ml) was measured by ELISA on the cells treated with 1 μg /
(2) 전 염증성 사이토카인 유도 분석결과(2) Results of proinflammatory cytokine induction assay
대청엽 추출물에 의한 전 염증성 사이토카인 유도 분석결과를 도 6에 개시하였다. 종양괴사인자 알파(tumor necrosis factor-α; TNF-α)는 주로 활성화된 대식세포에 의해 분비되며, 가장 중요한 역할은 면역세포의 조절이다. 또한 바이러스 복제를 억제하는 능력이 있는 것으로 알려져 있다. 인터루킨 6(Interleukin 6; IL-6는 B-세포를 활성화시켜 항체생산을 증가시켜 항원특이적 면역반응을 촉진하는 중요한 사이토카인이다. 인터페론(Interferon; IFN)은 세포의 조절물질로서 그 기능이 아주 다양하다. 예를 들면, 바이러스로부터의 세포보호, 조직배양에서나 골수에서의 세포분열 억제, T세포의 작용 조절, 자연면역세포(NK세포)의 기능 항진을 유도하여 식균작용을 상승시키고, 특수 암세포의 분열 억제하는 것으로 알려져 있다. 도 6에 개시한 바와 같이, Raw 264.7 세포에서 대청엽 추출물에 의해 전 염증성 사이토카인인 IL-6 및 IFN-β가 강하게 유도되는 것을 확인할 수 있다.
The results of the proinflammatory cytokine induction assay by the extract of Daechungchon were shown in Fig. Tumor necrosis factor-α (TNF-α) is mainly secreted by activated macrophages, the most important role being the regulation of immune cells. It is also known to have the ability to inhibit viral replication. Interleukin 6 (IL-6) is an important cytokine that stimulates B-cell activation and stimulates antigen-specific immune responses by increasing antibody production. Interferon (IFN) For example, cell protection from viruses, inhibition of cell division in bone marrow, suppression of T cell function, and hyperactivity of naturally-occurring immune cells (NK cells), thereby increasing phagocytosis, As shown in Fig. 6, it can be confirmed that IL-6 and IFN- [beta], which are proinflammatory cytokines, are strongly induced by extracts of Miscanthidae from Raw 264.7 cells.
실시예Example 4. 4. 대청엽Big shrine leaf 추출물에 의한 By extract H1N1H1N1 및 And H5N2H5N2 바이러스 감염 억제 분석 Virus Infection Inhibition Analysis
(1) 분석방법(1) Analysis method
MDCK 세포주에 1㎍/㎖의 대청엽 추출물과 1.0 MOI(multiplicity of infection) 바이러스를 동시에 처리하고 24시간 후, 세포독성을 측정하기 위하여 10㎕의 Ez-Cytox 시약을 처리하고 12시간 동안 배양하고 450nm에서 흡광도를 측정하였다.
MDCK cell line was treated with 1 쨉 g / ml of extract of Daechung leaf and 1.0 MOI (multiplicity of infection) at the same time. After 24 hours, 10 쨉 l of Ez-Cytox reagent was treated for 12 hours to measure cytotoxicity, The absorbance was measured.
(2) 분석결과(2) Analysis results
대청엽 추출물에 의한 H1N1 및 H5N2 바이러스 감염에 의한 세포 생존도를 분석한 결과, H1N1 및 H5N2 바이러스 감염에 의해 세포 생존률이 약 20%로 감소하였으며, 1㎍/㎖의 대청엽 추출물과 1 MOI 바이러스를 동시에 처리한 군의 경우, 세포 생존률이 약 50%로 나타나, 바이러스 감염에 의한 세포 생존률의 감소를 막아주는 것을 확인하였다(도 7).
Analysis of cell viability by H1N1 and H5N2 virus infection by extracts of Daechungpo leaves revealed that cell viability was reduced to about 20% by H1N1 and H5N2 virus infection and 1 μg / In the group treated at the same time, the cell survival rate was about 50%, and it was confirmed that the decrease in the cell survival rate due to the virus infection was prevented (FIG. 7).
제조예Manufacturing example 1. 주사제 1. Injection
상기 실시예 1에서 제조한 대청엽 추출물: 100㎎The extract of the macrocarpa extract prepared in Example 1: 100 mg
소듐 메타비설파이트: 3.0㎎Sodium metabisulfite: 3.0 mg
메틸파라벤: 0.8㎎Methylparaben: 0.8 mg
프로필파라벤: 0.1㎎Propyl paraben: 0.1 mg
주사용 멸균 증류수: 적량Sterile sterilized distilled water for injection:
상기 성분을 혼합하고 통상의 방법으로 최종 부피가 2 ㎖이 되도록 제조하여, 앰플에 충전하고 멸균하여 주사제를 제조하였다.
The above ingredients were mixed and made into a final volume of 2 ml by a conventional method, filled in an ampoule and sterilized to prepare an injection.
제조예Manufacturing example 2. 정제 2. Refining
상기 실시예 1에서 제조한 대청엽 추출물: 200㎎The extract of the macrocarpal extract prepared in Example 1: 200 mg
감자 전분: 100㎎Potato starch: 100 mg
락토오스: 100㎎Lactose: 100 mg
콜로이드성 규산: 16㎎Colloidal silicic acid: 16 mg
스테아린산 마그네슘: 적량Magnesium stearate:
통상의 정제 제조방법에 따라 상기 성분을 혼합하고 타정하고 정제를 제조하였다.
The ingredients were mixed and tableted according to a conventional tablet preparation method to prepare tablets.
제조예Manufacturing example 3. 3. 캡슐제Capsule
상기 실시예 1에서 제조한 대청엽 추출물: 100㎎The extract of the macrocarpa extract prepared in Example 1: 100 mg
유당: 50㎎Lactose: 50 mg
전분: 50㎎Starch: 50 mg
탈크: 2㎎Talc: 2 mg
스테아린산 마그네슘: 적량Magnesium stearate:
통상의 캡슐 제조방법에 따라 상기 성분을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.
The above components were mixed according to a conventional capsule manufacturing method and filled in gelatin capsules to prepare capsules.
제조예Manufacturing example 4. 4. 환제Pill
상기 실시예 1에서 제조한 대청엽 추출물: 120㎎The extract of the macrocarpal extract prepared in Example 1: 120 mg
옥수수 전분: 100㎎Corn starch: 100 mg
멸균 증류수: 적량Sterilized distilled water: suitable amount
상기 성분을 혼합하고, 통상의 환제 제조방법에 따라 적절한 크기를 갖는 구형으로 제환하여 환제를 제조하였다.
The above ingredients were mixed and pelletized to spheres of appropriate size according to conventional pellet manufacturing methods to produce pellets.
제조예Manufacturing example 5. 건강 기능 식품 5. Health functional foods
1) 건강 음료1) Health drinks
올리고당(2%), 액상과당(0.5%), 설탕(2%), 식염(0.5%), 물(75%) 등의 음료 재료에 상기 실시예 1에서 제조된 대청엽 추출물을 적량 혼합하여, 살균함으로써 음료를 제조하였다. The extracts prepared in Example 1 were mixed in an appropriate amount to drink materials such as oligosaccharide (2%), liquid fructose (0.5%), sugar (2%), salt (0.5% The beverage was prepared by sterilization.
2) 기능성 식품2) Functional food
상기 실시예 1에서 제조한 대청엽 추출물을 각종 비타민 및 미네랄 함유 기능성 식품에 적량 혼합하여 대청엽 추출물이 함유된 기능성 식품을 제조하였다.
The functional food containing the extract of Miscanthus multiflora was prepared by mixing the extract of the Miscanthus sinensis extract prepared in Example 1 with various vitamins and minerals in a proper amount.
제조예Manufacturing example 6. 사료 조성물 6. Feed composition
동물용 배합 사료에 상기 실시예 1에서 제조된 대청엽 추출물을 적량 혼합하여, 사료 조성물을 제조한 후, 펠렛화 및 과립화하였다.The animal feed composition was mixed with the appropriate amount of the extract prepared in Example 1 to prepare a feed composition, which was then pelletized and granulated.
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| KR102034313B1 (en) * | 2018-02-20 | 2019-10-18 | 한국식품연구원 | Composition for Preventing, Improving or Treating of muscular disease containing Isatis indigotica Fortune extract |
| KR102304498B1 (en) * | 2020-03-05 | 2021-09-23 | 남종현 | Composition for anti-transmissible gastroenteritis virus (tgev) and pharmaceutical composition including the same |
| KR102467386B1 (en) * | 2021-05-14 | 2022-11-16 | 엠테라파마 주식회사 | Composition for preventing, improving or treating inflammatory bowel disease comprising herbal extracts of isatidis folium |
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| CN104206678A (en) * | 2014-08-25 | 2014-12-17 | 青岛众泰禽业专业合作社 | Manufacture method of feed additive of Isatis indigotica Fortune extract |
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Non-Patent Citations (2)
| Title |
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| The American Journal of Chinese Medicine, vol.41, no.4, pp.957-969 (2013.)* |
| 중국특허공개공보 104206678(2014.12.17.)* |
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