KR20140037899A - Immune enhancing and antiviral composition comprising extract of salvia miltiorrhiza bunge - Google Patents
Immune enhancing and antiviral composition comprising extract of salvia miltiorrhiza bunge Download PDFInfo
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- KR20140037899A KR20140037899A KR1020140008947A KR20140008947A KR20140037899A KR 20140037899 A KR20140037899 A KR 20140037899A KR 1020140008947 A KR1020140008947 A KR 1020140008947A KR 20140008947 A KR20140008947 A KR 20140008947A KR 20140037899 A KR20140037899 A KR 20140037899A
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Abstract
Description
본 발명은 면역 증강 및 항바이러스 조성물에 관한 것으로, 더욱 상세하게는 의약, 건강기능식품 및 사료 조성물 등으로 이용될 수 있는 단삼 추출물을 포함하는 면역 증강 및 항바이러스 조성물에 관한 것이다.
The present invention relates to an immunostimulatory and antiviral composition, and more particularly, to an immunostimulatory and antiviral composition comprising a ginseng extract which can be used as a medicament, a health functional food, a feed composition or the like.
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.
생체내 선천적 방어 면역시스템 중 최근 가장 중요시되는 연구 분야는 인터페론(interferon)에 의해 유도되는 선천성 면역분야이며, 이러한 인터페론 매개 면역의 활성화는 다양한 전염병 병원체에 대한 근본적인 예방 방법이 될 수 있다. 따라서 인터페론 활성화 기전 연구 및 인터페론을 유도시킬 수 있는 면역조절제제의 개발 연구가 활발하다. 또한, 병원체의 감염에 따른 선천 면역의 방어기작의 하나로 면역 인자(염증 사이토카인)가 분비되고, 이들 인자들에 의해 염증반응이 유발되어 병원체에 대한 방어가 이루어진다. 따라서 적정한 수준의 염증반응 유도 역시 다양한 전염병 병원체에 대한 예방 및 치료 방법이 될 수 있으며, 이를 유도시킬 수 있는 면역증강제제의 개발 연구가 필요하다.Among the in vivo innate defense immune systems, the most important research field is the field of congenital immunity induced by interferon. Such 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. In addition, immune factors (inflammatory cytokines) are secreted as a defense mechanism against congenital immune diseases caused by pathogenic infections, and inflammatory responses are induced by these factors to protect against pathogens. Therefore, induction of inflammatory response at an appropriate level may be a preventive and therapeutic method for various infectious disease pathogens, and development of an immunity enhancer that can induce the inflammatory response is needed.
바이러스(virus)란 라틴어로 독성물질을 의미하며, 세균여과지(0.22㎛)를 통과하는 일군의 감염형 병원성 입자를 의미한다. 바이러스는 숙주세포의 종류에 따라 박테리오 파지, 식물 바이러스, 동물 바이러스로 분류하기도 하며, 핵산의 종류에 따라 DNA 바이러스, RNA 바이러스로 분류할 수 있다. 최근 신종플루, 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 means a toxic substance in Latin and refers to a group of infectious pathogenic particles that pass through bacterial filter paper (0.22 μm). Viruses can be classified as bacteriophages, plant viruses, and animal viruses according to the type of host cells. 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 위해서 가장 좋은 방법은 백신접종이다. 그럼에도 불구하고 RNA 바이러스에 의한 질병의 경우, 대체적으로 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 생체내 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. Nevertheless, in the case of diseases caused by RNA viruses, the efficiency of vaccines due to the generation of many virus 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.
인플루엔자 바이러스의 증식을 억제하는 대표적인 항바이러스 제제로는 아만타딘(amantadine)과 리만타딘(rimantadine)이 있으나, 이들 두 가지 항바이러스 제제들은 혈청형 A형 인플루엔자 바이러스에만 효과적이며, M2 단백질이 없는 혈청형 B형 인플루엔자 바이러스에는 효과가 없는 것으로 확인되었다. 또한 아만타딘과 리만타딘은 사용 시 인플루엔자 바이러스 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 인플루엔자 바이러스와 혈청형 B형 인플루엔자 바이러스에 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. 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, 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 다가혼합 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, 수포성 구내염 바이러스(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, the main control method of vesicular stomatitis virus is the complete treatment of the disease, so as with foot-and-mouth disease, prevention and blocking prevention and eradication of susceptible domestic livestock are the best methods.
이러한 상황하에서, 최근 해외 및 국내에서 기존의 항바이러스 제제의 단점을 극복하고자 많은 노력들이 이루어지고 있으며, 그 노력 중의 하나로 국내에서는 생약 추출물 및 식물 추출물을 대상으로 항바이러스 효능에 대한 연구가 활발히 진행중이다.Under these circumstances, many efforts have recently been made to overcome the disadvantages of existing antiviral agents both in foreign countries and domestic ones. As one of the efforts, researches on antiviral efficacy of herbal medicine extracts and plant extracts are actively underway in Korea .
한편, 단삼(Salvia miltiorrhiza BUNGE)은 꿀풀과(Labiatae)에 속하는 다년초로, 뿌리에 함유된 것으로 알려진 성분은 바이칼린(baicalin) 등의 플라보노이드 성분(flavonoids), 이소탄신온(isotanshione) I, 탄신온(tanshinone) I, 탄신온 IIA, 탄신온 IIB, 크립토탄신온(cryptotanshinone) 등의 수종의 디테르펜 성분(diterpenes), 올레안산(oleanoic acid) 등의 트리테르펜 성분(triterpenes), 로즈마린산(rosmarinic acid) 등의 페닐프로판노이드 성분(phenylpropanoids), 살비아놀산(salvianolic acid), 리소스퍼믹산(lithospermic acid) B 등의 리그난 성분(lignans) 등이 있다. 이중 디테르펜계 성분이 가장 많이 함유되어 있는데 이소탄신온 I, 탄신온 I, 탄신온 IIA, 탄신온 IIB, 크립토탄신온 등은 단삼에만 함유되어 있는 성분이다. 단삼을 이용한 종래기술은 간질환 치료 및 예방을 위한 조성물, 간섬유화 또는 간경화증 예방 및 치료용 조성물과 같이 간세포 등에 미치는 영향에 대한 기술이 주를 이루고 있다.Salvia miltiorrhiza BUNGE belongs to the perennial plant belonging to the family Lamiaceae. The components known to be contained in the roots include flavonoids such as baicalin, isotanshione I, triterpenes such as diterpenes, oleanolic acid and triterpenes such as tanshinone I, cinnamon IIA, cinnamon IIB and cryptotanshinone, rosmarinic acid, And lignans such as phenylpropanoids, salvianolic acid, and lithospermic acid B, and the like. Among the diterpene compounds, isotansinone I, carcinogen I, carcinogen IIA, carcinogen IIB, and cryptotancinone are the components contained only in radish. Background Art [0002] The prior art using Dansamp is mainly directed to a composition for the treatment and prevention of liver diseases, and to an effect on hepatocytes such as a composition for prevention and treatment of liver fibrosis or liver cirrhosis.
이에, 기존의 항바이러스 제제의 단점을 극복하여 독성 및 부작용이 거의 없이 우수한 면역 증강 효과 및 항바이러스 활성을 발휘할 수 있는 단삼 추출물을 이용한 조성물에 대한 요구가 여전히 존재하는 실정이다.
Accordingly, there is still a need for a composition using the extract of Dansamp extract which can overcome the disadvantages of existing antiviral agents and exert excellent immune enhancing effect and antiviral activity with little toxicity and side effects.
본 발명자들은 단삼의 추출물이 선천 면역의 주요 세포인 대식 세포를 활성화시켜 다양한 RNA 바이러스의 증식을 억제시킬 수 있음을 확인하여 본 발명을 완성하였다. 본 발명은 독성 및 부작용이 거의 없어 장기간 안정성을 확보하면서, 각종 바이러스 또는 세균 감염성 질병의 예방 및 치료, 약학 조성물, 건강기능식품 또는 가축 사료 조성물 등에 효과적으로 이용될 수 있는 단삼 추출물을 포함하는 면역 증강 및 항바이러스 조성물 및 동물의 면역 증강 및 항바이러스 활성 증강 방법을 제공하는 것을 그 목적으로 한다.
The inventors of the present invention completed the present invention by confirming that the extract of Dansamp was able to inhibit the proliferation of various RNA viruses by activating macrophages which are the main cells of innate immunity. The present invention relates to a pharmaceutical composition for preventing and treating various viruses or bacterial infectious diseases while securing long-term stability with little toxicity and side effects, It is an object of the present invention to provide an antiviral composition and a method for enhancing immunity and antiviral activity of an animal.
상기 과제를 해결하기 위한 본 발명의 일 측면은 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 약학 조성물을 제공한다.One aspect of the present invention for solving the above-mentioned problems provides a pharmaceutical composition for immune enhancement and antiviral treatment, which comprises extracts of Panax ginseng.
또한, 본 발명의 다른 일 측면은 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 건강기능식품을 제공한다.In addition, another aspect of the present invention provides a health functional food for immune enhancement and antiviral activity, which comprises extracts of Panax ginseng.
또한, 본 발명의 또 다른 일 측면은 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 사료 조성물을 제공한다.Yet another aspect of the present invention provides an immunostimulating and antiviral feed composition comprising a root extract.
또한, 본 발명의 또 다른 일 측면은 단삼 추출물을, 인간을 제외한 면역 증강이 필요한 동물에 투여하는 것을 포함하는 동물의 면역 증강 및 항바이러스 활성 증강을 제공한다.
Yet another aspect of the present invention provides immune enhancement and antiviral activity enhancement of an animal, comprising administering the extract to a subject in need thereof.
본 발명에 따르면 우수한 면역 증강 효능을 발휘하는 단삼 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 Radix Salviae which exerts excellent immunity enhancing effect, and can contribute to the enhancement of immunity.
또한, 본 발명에 따른 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 조성물은 독성이나 부작용을 거의 일으키지 않으므로 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있다.In addition, the immunosuppressive and antiviral composition comprising the extract of Dansamp extract according to the present invention scarcely causes toxicity or side effects, so that it can be safely used for prophylactic purposes for a long period of time.
따라서, 본 발명에 따른 조성물은 박테리아 및 바이러스 감염성 질환의 예방 및 치료용으로 적용될 수 있고, 면역이 저하되거나 면역이 억제된 환자의 면역력 증강 및 조절을 위한 약학 조성물이나 건강기능식품, 및 동물의 항질병 강화를 목적으로 하는 면역증강용 사료 조성물 등으로 광범위하게 이용될 수 있다.Therefore, the composition according to the present invention can be applied for the prevention and treatment of bacterial and viral infectious diseases, and can be applied to pharmaceutical compositions and health functional foods for immunity enhancement and control of immunocompromised or immunosuppressed patients, An animal feed composition for immunity enhancement aimed at enhancing disease, and the like.
또한, 본 발명에 따르면 단삼 추출물을 면역 증강에 필요한 동물에 투여함으로써 동물의 면역을 효과적으로 안전하게 증강시킬 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 Dansampus to an animal necessary for immunity enhancement.
도 1은 본 발명에 실시예에 따른 단삼 추출물의 세포 독성 실험결과를 나타낸다.
도 2는 본 발명의 실시예에 따른 단삼 추출물의 PR8-gfp(Swine influenza virus)에 대한 항바이러스 활성 실험결과를 나타낸다.
도 3은 본 발명의 실시예에 따른 단삼 추출물의 VSV-gfp(Vesicular stomatitis virus)에 대한 항바이러스 활성 실험결과를 나타낸다.
도 4는 본 발명의 실시예에 따른 단삼 추출물의 NDV-gfp(Newcastle disease virus)에 대한 항바이러스 활성 실험결과를 나타낸다.
도 5는 본 발명의 실시예에 따른 단삼 추출물의 농도에 따른 항바이러스 활성 실험결과를 나타낸다.
도 6은 본 발명의 실시예에 따른 단삼 추출물에 의한 전염증성 사이토카인 유도 실험결과를 나타낸다.
도 7은 본 발명의 실시예에 따른 단삼 추출물에 의한 인터페론-β 분비 유도 실험결과를 나타낸다.
도 8은 본 발명의 실시예에 따른 in - vivo 동물 실험에 있어서 단삼 추출물에 의한 인플루엔자 바이러스 감염 억제 실험결과를 나타낸다.FIG. 1 shows the cytotoxicity test results of the extract of Radix Salviae Radix according to the example of the present invention.
FIG. 2 shows the results of the antiviral activity test on the PR8-gfp (Swine influenza virus) of the extracts of Rana ginseng according to the embodiment of the present invention.
FIG. 3 shows the results of an antiviral activity test for VSV-gfp (Vesicular stomatitis virus) of the extract of Rana ginseng according to an embodiment of the present invention.
FIG. 4 shows the results of an antiviral activity test for the NDV-gfp (Newcastle disease virus) of the extract of Rana ginseng according to an embodiment of the present invention.
FIG. 5 shows the results of the antiviral activity test according to the concentration of the ginseng extract according to an embodiment of the present invention.
FIG. 6 shows the results of induction of proinflammatory cytokines by the extracts of Panax ginseng according to an embodiment of the present invention.
FIG. 7 shows experimental results of induction of interferon-beta secretion by the extract of Panax ginseng according to the example of the present invention.
Figure 8 in accordance with an embodiment of the present invention shows an inhibition of influenza virus infection experiment result by the Salvia miltiorrhiza extract in vivo animal experiments.
이하, 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 본 발명의 기술적 사상을 용이하게 실시할 수 있을 정도로 상세히 설명하기 위하여, 본 발명의 바람직한 실시예를 설명하기로 한다.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings.
본 발명은 선천 면역의 주요 세포인 대식 세포를 활성화시켜 다양한 RNA 바이러스의 증식을 억제시킬 수 있는 단삼 추출물을 이용함으로써 면역 증강 효과를 나타내는 것을 특징으로 한다.The present invention is characterized by exhibiting an immunostimulating effect by using a ginseng extract which can activate macrophages, the main cells of innate immunity, and inhibit the proliferation of various RNA viruses.
본 발명에 이용되는 단삼 추출물은 일반적으로 천연물 추출에 사용되는 다양한 용매를 이용한 추출에 의하여 형성될 수 있으며, 예를 들어, 열수 추출, 알코올 추출 및 주정 추출로 이루어진 군으로부터 선택되는 일 이상의 방법에 의하여 형성될 수 있다.The extract of Dansamp for use in the present invention can be generally formed by extraction using various solvents used for extracting natural products, for example, by one or more methods selected from the group consisting of hot water extraction, alcohol extraction and alcohol extraction .
일 실시예에서, 단삼 추출물은, 증류수를 첨가하는 단계; 60~130℃, 바람직하게는 약 105℃의 온도에서 30~240분 동안, 바람직하게는 약 150분 동안 열수 추출하는 단계; 및 여과하는 단계를 거쳐 형성될 수 있다.In one embodiment, the extract is prepared by adding distilled water; Hydrothermal extraction at a temperature of 60 to 130 캜, preferably about 105 캜, for 30 to 240 minutes, preferably about 150 minutes; And it may be formed through a filtration step.
단삼 추출물은 각종 바이러스, 예를 들면 인플루엔자 바이러스, 뉴캐슬병 바이러스, 수포성 구내염 바이러스 등에 대하여 우수한 항바이러스 활성을 나타내고, 면역 인자 유도능을 강하게 나타내며, 개체의 선천 면역을 증가시켜 인플루엔자 바이러스의 감염 및 증식을 억제하는 효과를 발휘한다.Dansamp extract has excellent antiviral activity against various viruses such as influenza virus, Newcastle disease virus and vesicular stomatitis virus. It strongly shows the immunity factor induction ability and increases the innate immune status of the individual, thereby increasing the infection and proliferation of influenza virus Suppressing effect.
또한, 단삼 추출물은 세포 독성이나 부작용을 거의 나타내지 않으므로 장기간 복용시에도 안심하고 사용할 수 있다.In addition, the extract of Dansamp shows little cytotoxicity or side effects, so it can be safely used even when taken for a long time.
본 발명에 있어서는 이와 같은 효능을 발휘하는 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 약학 조성물, 면역 증강 및 항바이러스용 건강기능식품 또는 면역 증강 및 항바이러스용 사료 조성물 등에 적용할 수 있다.The present invention can be applied to a pharmaceutical composition for immunity enhancement and antiviral activity, a health functional food for immunity enhancement and antivirus, or a feed composition for immunity enhancement and antivirus, which comprises the extract of Dansamp extract exhibiting such efficacy.
본 발명의 일 측면은 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 약학 조성물에 관한 것이다.One aspect of the present invention relates to a pharmaceutical composition for immunostimulating and antivirals that comprises extracts of Panax ginseng.
일 실시예에서, 본 발명의 면역 증강 및 항바이러스용 약학 조성물은 약학적으로 허용가능한 담체, 부형제 또는 희석제를 더 포함한다.In one embodiment, the pharmaceutical compositions for the immunostimulant and antiviral of the present invention further comprise a pharmaceutically acceptable carrier, excipient or diluent.
본 발명에 이용될 수 있는 약학적으로 허용가능한 담체, 부형제 또는 희석제는 본 발명의 효과를 해하지 않는 한 특히 제한되지 않으며, 예를 들어 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제, 윤활제, 감미제, 방향제, 보존제 등을 포함할 수 있다.Pharmaceutically acceptable carriers, excipients or diluents which can be used in the present invention are not particularly limited so long as they do not impair the effects of the present invention, for example fillers, extenders, binders, wetting agents, disintegrants, surfactants, lubricants, Sweeteners, fragrances, preservatives and the like.
약학적으로 허용가능한 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.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 Silicates, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, propylene glycol, polyethylene glycol, vegetable oil, injectable Ester, utopsol, macrogol, tween 61, cacao butter, lauridge, etc. are mentioned.
본 발명의 면역 증강 및 항바이러스용 약학 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁액, 에멀젼, 시럽제, 에어로졸, 외용제, 좌제 및 주사제로 이루어진 군으로부터 선택되는 형태일 수 있다.The pharmaceutical composition for immunostaining and antiviral of the present invention may be in the form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and injections.
약학 조성물의 제제화 방법은 기술분야에 공지된 통상의 방법에 따라 수행될 수 있으며, 특히 제한되지 않는다.The method of formulating the pharmaceutical composition may be carried out according to conventional methods known in the art, and is not particularly limited.
본 발명의 면역 증강 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 compositions for immunological enhancement and antiviral therapy of the present invention can be administered orally or parenterally. The dosage may be appropriately adjusted according to the age, sex, weight, condition, degree of disease, drug form, 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 면역 증강 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 methods of administration, dosages, route of administration, constituents, etc. of the pharmaceutical composition for immunoenhancing and antiviral of the present invention can be suitably selected from conventional techniques known in the art.
본 발명의 면역 증강 및 항바이러스용 약학 조성물은 박테리아 감염성 질병 또는 바이러스 감염성 질병의 예방 및 치료에 이용될 수 있다.The pharmaceutical composition for immunostaining and antiviral of the present invention can be used for the prevention and treatment of bacterial infectious diseases or viral infectious diseases.
특히, 본 발명의 면역 증강 및 항바이러스용 약학 조성물은 독성이나 부작용을 거의 갖지 않는 단삼 추출물을 유효성분으로 함유하므로, 예방 목적으로 장기간 복용시에도 안심하고 사용할 수 있다.In particular, the pharmaceutical composition for immune enhancement and antiviral treatment of the present invention contains the extract of Rana ginseng, which has little or no toxicity or side effects, as an active ingredient, so that it can be safely used for prophylactic purposes for a long period of time.
일 실시예에서, 본 발명의 면역 증강 및 항바이러스용 약학 조성물은 유효 성분으로 단삼 추출물 외에 다른 약학적 활성 성분을 함께 포함하거나, 또는 다른 유효 성분을 포함하는 약학 조성물과 혼합되어 이용될 수 있다.In one embodiment, the pharmaceutical composition for immuno-enhancement and antiviral use of the present invention may be used as an active ingredient in combination with a pharmacologically active ingredient in addition to the extract, or may be mixed with a pharmaceutical composition containing another active ingredient.
예를 들어, 본 발명의 면역 증강 및 항바이러스용 약학 조성물은 암, 특히 전립선, 결장, 폐, 유방, 난소, 두경부, 외음부, 방광 및 뇌암, 신경교종뿐 아니라 비전염성 만성 질병의 예방 또는 치료용 약학 조성물, 바이러스 및 세균 감염성 질병의 예방 또는 치료용 약학 조성물, 또는 자가면역 질병의 치료용 약학 조성물 등과 함께 사용될 수 있다.For example, the pharmaceutical compositions for immuno-enhancement and antiviral use of the present invention can be used for prevention or treatment of cancer, particularly prostate, colon, lung, breast, ovary, head and neck, vulvar and bladder cancer, glioma as well as non-communicable chronic diseases A pharmaceutical composition for the prevention or treatment of viral and bacterial infectious diseases, or a pharmaceutical composition for the treatment of autoimmune diseases and the like.
본 발명의 다른 일 측면은 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 건강기능식품에 관한 것이다.Another aspect of the present invention relates to a health functional food for immune-enhancing and antiviral treatment comprising extracts of Panax ginseng.
일 실시예에서, 본 발명의 면역 증강 및 항바이러스용 건강기능식품은 식품학적으로 허용가능한 식품 보조 첨가제를 더 포함한다.In one embodiment, the immunomodulatory and antiviral health functional food of the present invention further comprises a pharmaceutically acceptable food-aid additive.
본 발명에 이용될 수 있는 식품학적으로 허용가능한 식품 보조 첨가제는 포도당, 과당, 말토스, 슈크로스, 덱스트린, 시클로덱스트린과 같은 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알코올과 같은 천연 탄수화물, 타우마틴, 스테비아 추출물 등의 천연 향미제, 사카린, 아스파르탐 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.Food acceptable acceptable food supplement additives which can be used in the present invention include sugars such as glucose, fructose, maltose, sucrose, dextrin, cyclodextrin and natural carbohydrates such as sugar alcohols such as xylitol, sorbitol, erythritol, tau Natural flavors such as martin and stevia extract, synthetic flavors such as saccharin, aspartame, colorants, pectic acid or salts thereof, alginic acid or salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin , Alcohols, carbonating agents, and the like, but are not limited thereto.
본 발명의 면역 증강 및 항바이러스용 건강기능식품은 분말, 과립, 정제, 캡슐, 캔디, 츄잉검, 젤리 및 음료로 이루어진 군으로부터 선택되는 형태일 수 있다.The immunosuppressive 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.
면역 증강 및 항바이러스용 건강기능식품 중의 단삼 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.The content of the ginseng extract in the immunoenhancing and antiviral health functional food may be appropriately selected in consideration of the form, flavor, taste, etc. of the food, and may be, for example, in the range of 0.01 to 30 wt% Lt; / RTI >
본 발명의 면역 증강 및 항바이러스용 건강기능식품의 형태, 조성 및 제조방법 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다. It is apparent to those skilled in the art that the form, composition and manufacturing method of the immunoenhancing and antiviral health functional food of the present invention can be suitably selected from conventional techniques known in the art.
본 발명의 또 다른 일 측면은 단삼 추출물을 포함하는 면역 증강 및 항바이러스용 사료 조성물에 관한 것이다.Another aspect of the present invention relates to a feed composition for immunostimulating and antivirals comprising the extract of Panax ginseng.
면역 증강 및 항바이러스용 사료 조성물 중의 단삼 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.The content of the ginseng extract in the feed composition for immunity enhancement and antiviral treatment may 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, preferably 0.1 To 80% by weight.
본 발명의 면역 증강 및 항바이러스용 사료 조성물은 기술분야에 공지된 사료 제조방법에 따라 제조될 수 있으며, 예를 들어, 각종 사료 원료 또는 배합사료와 본 발명의 단삼 추출물을 혼합한 후, 추가적인 가공 공정, 예를 들어 펠렛 형태로의 성형 또는 과립 등의 형태로의 절단 단계 등을 더 수행함으로써 제조될 수 있다.The feed composition for immune enhancement and antiviral of the present invention may be prepared according to a feed preparation method known in the art, for example, after mixing various feed ingredients or blended feed with the extract of the present invention, and further processing It can be produced by further performing a process, for example, molding into pellets or cutting into granules and the like.
본 발명의 면역 증강 및 항바이러스용 사료 조성물의 구성성분, 조성, 제조방법, 급여방법 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다.It will be apparent to those skilled in the art that the composition, composition, manufacturing method, feeding method, etc. of the immunostimulatory and antiviral feed composition of the present invention can be suitably selected from conventional techniques known in the art.
본 발명의 또 다른 일 측면은 단삼 추출물을, 인간을 제외한 면역 증강이 필요한 동물에 투여하는 것을 포함하는 동물의 면역 증강 및 항바이러스 활성 증강에 관한 것이다.Another aspect of the present invention relates to the enhancement of immunity and antiviral activity of an animal, which comprises administering a root extract to an animal in need of immune enhancement other than human.
본 발명의 동물의 면역 증강 및 항바이러스 활성 증강에 있어서, 단삼 추출물의 투여량, 투여경로, 투여시기 등은 기술분야에 공지된 통상의 기술로부터 적절히 선택될 수 있음이 통상의 기술자에게 명백하다. It is apparent to those skilled in the art that the dose, route of administration, administration time, etc. of the extract of Panax notoginseng can be suitably selected from conventional techniques known in the art in enhancing the immunity and antiviral activity of the animal of the present invention.
이와 같이 본 발명에 있어서는 대식 세포를 활성화시켜 다양한 RNA 바이러스의 증식을 억제하고, 독성이나 부작용이 거의 없는 단삼 추출물을 약학 조성물, 건강기능식품 또는 사료 조성물 등에 이용함으로써, 면역 증강 및 조절, 동물의 항질병 강화 효과를 발휘할 수 있으며, 나아가 동물의 면역 증강 및 항바이러스 활성 증강을 제공할 수 있다.As described above, in the present invention, it is possible to prevent the growth of various RNA viruses by activating macrophages, and by using the extract of Ganoderma lucidum, which has little toxicity or side effects, in a pharmaceutical composition, a health functional food or a feed composition, Can exert a disease-strengthening effect, and can further provide an animal with enhanced immunity and antiviral activity.
이하, 본 발명을 실시예에 의하여 더욱 상세하게 설명한다. 그러나 하기 실시예는 본 발명을 예시하는 것일 뿐이며, 본 발명의 범위가 하기 실시예에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail by way of examples. However, the following examples are illustrative of the present invention, and the scope of the present invention is not limited to the following examples.
[[ 실시예Example ]]
1. 약재의 선정 1. Selection of medicinal materials
(1) 약재의 수집(1) Collection of medicinal materials
상한론, 금궤요략, 방약합편 등에 기재된 처방 및 그 처방에 포함된 단미제 등을 대상으로 항균 활성 및 면역 증강 효능이 있다고 보고된 약재들을 수집하여 실험에 사용하였다.The antimicrobial activity and antimicrobial activity of the antimicrobial activity and antimicrobial activity of antimicrobial agents contained in the antimicrobial agent, antimicrobial activity, and antimicrobial activity were investigated.
(2) 생약 추출물의 제조(2) Preparation of herbal medicine extract
선발된 생약 추출물은 10배수의 증류수를 첨가하여 105℃에서 150분간 열수 추출하고, 0.45㎛로 1차 여과한 후, 0.22㎛로 최종 여과하여 침전물을 제거하였다. 이어서, pH를 7.0으로 조정하여 1.5㎖ Ep-튜브에 1㎖씩 분주하고 -20℃에서 저장하여 모든 실험에 사용하였다.
The extracted herbal extracts were subjected to hot water extraction at 105 ° C. for 150 minutes by adding 10 times of distilled water, followed by primary filtration at 0.45 μm and final filtration at 0.22 μm to remove precipitates. 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 all experiments.
2. 마우스 2. Mouse 대식세포주를Macrophage cell line 이용한 Used 단삼Sansam 추출물의 세포 독성 Cytotoxicity of extract
: 단삼 추출물의 세포 독성을 확인하기 위하여 안전성 실험을 하기와 같이 수행하였다.
: In order to confirm the cytotoxicity of the extract, the safety experiment was carried out as follows.
(1) 세포 독성 실험방법(1) Cytotoxicity test method
1) 마우스 대식세포주인 Raw 264.7을 배양하여 실험에 사용하였다.1) Mouse macrophage cell line Raw 264.7 was cultured and used in the experiment.
2) 12-웰 TC 플레이트에 세포를 배양한 후, 1% FBS가 첨가된 DMEM에 상기에서 제조된 단삼 추출물(pH 조정)을 각각 0.3%, 0.5%, 1% 및 2%의 농도로 첨가하여 처리하였다.2) Cells were cultured on a 12-well TC plate, and then added with the concentrations of 0.3%, 0.5%, 1% and 2%, respectively, of the radish extract (pH adjusted) prepared above in DMEM supplemented with 1% FBS Respectively.
3) 음성 대조군(Negative control)은 1% FBS가 첨가된 DMEM만을 처리하였다.3) Negative control was treated with DMEM supplemented with 1% FBS.
4) 22~24시간 후 0.4% 트립판 블루(Tryphan Blue)로 염색하여 세포 사멸을 확인하였다.
4) After 22-24 hours, the cells were stained with 0.4% tryphan blue to confirm apoptosis.
(2) 세포 독성 실험결과(2) Cytotoxicity test results
도 1에 세포 독성 실험 결과를 단삼 추출물의 농도별 세포 생존율로 나타낸다. 도 1로부터 확인할 수 있는 바와 같이, 단삼 추출물은 2% 이상의 농도에서도 Raw cell에 대해 세포 독성을 나타내지 않았다.
The results of the cytotoxicity test are shown in Fig. 1 as the cell survival rate by the concentration of Radix Salviae Radix extract. As can be seen from FIG. 1, the extracts of Dansamp showed no cytotoxicity against raw cells even at a concentration of 2% or more.
3. 마우스 3. Mouse 대식세포주를Macrophage cell line 이용한 Used 단삼Sansam 추출물의 항바이러스 활성 실험 Antiviral Activity Test of Extracts
: 다양한 RNA 바이러스, 즉 인플루엔자 바이러스(Influenza virus), 뉴캐슬병 바이러스(Newcastle disease virus), 수포성 구내염 바이러스(Vesicular stomatitis virus)에 대한 단삼 추출물의 항바이러스 활성 실험을 하기와 같이 수행하였다.
: Antiviral activity of various RNA viruses, namely Influenza virus, Newcastle disease virus and Vesicular stomatitis virus, was tested as follows.
(1) 항바이러스 활성 실험 방법(1) Antiviral activity test method
1) 마우스 대식세포주인 Raw 264.7을 키워 실험에 사용하였다(8 × 105 cell/well)1) Mouse macrophage cell line Raw 264.7 was used for the experiment (8 × 10 5 cells / well)
2) 12-웰 TC 플레이트에 세포를 배양한 후, 1% FBS가 첨가된 DMEM에 상기에서 제조된 단삼 추출물(pH 조정)을 1%의 농도로 첨가하여 처리하였다.2) Cells were cultured on a 12-well TC plate and treated with DMEM supplemented with 1% FBS at a concentration of 1%.
3) 음성 대조군은 1% FBS가 첨가된 DMEM만을 처리하였고, 양성 대조군(Positive control)은 Mouse IFN-B(500 units/㎖)를 처리하였다.3) Negative control group was treated with DMEM supplemented with 1% FBS, and positive control was treated with Mouse IFN-B (500 units / ml).
4) 단삼 처리 12시간 후, PR8-gfp(MOI:0.5), VSV-gfp(MOI:0.5), NDV-gfp(MOI:0.5)를 각각 접종하였다.4) After 12 hours of treatment, PR8-gfp (MOI: 0.5), VSV-gfp (MOI: 0.5) and NDV-gfp (MOI: 0.5) were inoculated respectively.
5) 접종하고 2시간 후 접종액을 제거하고, PBS로 3회 세척하고, 8~10시간 후 바이러스 감염 정도를 측정하였다.
5) After 2 hours of inoculation, the inoculum was removed, washed three times with PBS, and the degree of virus infection was measured after 8-10 hours.
(2) 항바이러스 활성 실험결과(2) Results of antiviral activity test
1) PR8-gfp(swine influenza virus) 실험결과1) PR8-gfp (swine influenza virus) test results
도 2에 PR8-gfp에 대한 항바이러스 활성 실험결과를 나타낸다. 도 2(a)는 감염 12시간 후의 GFP(green fluorescent protein) 형광이미지를 나타내고, 도 2(b)는 GFP 흡수도(absorbance)를 나타내며, 도 2(c)는 세포 생존율을 나타낸다(NC: 음성 대조군, PR8-gfp: 바이러스 감염군, 단삼: 단삼 추출물 처리군).Fig. 2 shows the results of the antiviral activity test for PR8-gfp. Figure 2 (a) shows GFP (green fluorescent protein) fluorescence image after 12 hours of infection, Figure 2 (b) shows GFP absorbance and Figure 2 (c) shows cell viability Control group, PR8-gfp: virus-infected group, and ginseng: ginseng extract-treated group).
도 2로부터, 단삼 추출물로 전처리한 결과 인플루엔자 바이러스 감염율이 현저하게 떨어졌으며, 세포 생존율도 바이러스 감염군에 비해 상승한 것을 확인할 수 있다.
From FIG. 2, it can be seen that the infection rate of influenza virus was remarkably lowered and the cell survival rate was also increased compared with the virus infection group as a result of pretreatment with the extract.
2) VSV-gfp(Vesicular stomatitis virus) 실험결과2) Results of VSV-gfp (Vesicular stomatitis virus)
도 3에 VSV-gfp에 대한 항바이러스 활성 실험결과를 나타낸다. 도 3(a)는 감염 12시간 후의 GFP 형광이미지를 나타내고, 도 3(b)는 감염 24시간 후의 바이러스 역가(viral titer) 그래프를 나타내며, 도 3(c)는 감염 24시간 후의 GFP 흡수도를 나타내고, 도 3(d)는 감염 24시간 후의 세포 생존율을 나타낸다(PC: 양성 대조군, VSV-gfp: 바이러스 감염군, 단삼: 단삼 추출물 처리군).FIG. 3 shows the results of the antiviral activity test for VSV-gfp. Figure 3 (a) shows the GFP fluorescence image after 12 hours of infection, Figure 3 (b) shows the viral titer graph after 24 hours of infection, and Figure 3 (c) (PC: positive control, VSV-gfp: virus-infected group, Dansamp: treatment with Panax ginseng extract). Fig. 3 (d) shows the cell survival rate after 24 hours of infection.
도 3으로부터 확인할 수 있는 바와 같이, 단삼 추출물로 전처리한 결과 수포성 구내염 바이러스 감염율이 현저하게 떨어졌으며, 바이러스 역가도 감소하였고, 세포 생존율도 바이러스 감염군에 비해 상승하였다.
As can be seen from FIG. 3, as a result of pretreatment with Radix Salviae Radix extract, the rate of vesicular stomatitis virus infection was remarkably decreased, the virus level was decreased, and the cell survival rate was also higher than the virus infection group.
3) NDV-gfp(newcastle disease virus) 실험결과3) NDV-gfp (newcastle disease virus)
도 4에 NDV-gfp에 대한 항바이러스 활성 실험결과를 나타낸다. 도 4(a)는 감염 12시간 후의 GFP 형광이미지를 나타내고, 도 4(b)는 감염 24시간 후의 GFP 흡수도를 나타내며, 도 4(c)는 감염 24시간 후의 세포 생존율을 나타낸다(NC: 음성 대조군, NDV-gfp: 바이러스 감염군, 단삼: 단삼 추출물 처리군).Figure 4 shows the results of the antiviral activity test for NDV-gfp. Fig. 4 (a) shows the GFP fluorescence image after 12 hours of infection, Fig. 4 (b) shows the GFP absorbance after 24 hours of infection, and Fig. 4 (c) Control group, NDV-gfp: virus-infected group, and ginseng: radish extract treatment group).
도 4로부터 확인할 수 있는 바와 같이, 단삼 추출물로 전처리한 결과 뉴캐슬병 바이러스 감염율이 현저하게 떨어졌으며, 세포 생존율도 바이러스 감염군에 비해 상승하였다.
As can be seen from FIG. 4, as a result of pretreatment with Radix Salviae Radix extract, the infection rate of Newcastle disease virus was remarkably decreased and the cell survival rate was also higher than that of virus infection group.
4. 4. 단삼Sansam 추출물의 농도에 따른 항바이러스 활성 실험 Antiviral activity test according to concentration of extract
: 단삼 추출물의 농도에 따른 항바이러스 활성을 확인하기 위하여 하기 실험을 수행하였다.
: The following experiment was carried out in order to confirm the antiviral activity according to the concentration of Radix Salviae Radix extract.
(1) 항바이러스 활성 실험방법(1) Antiviral activity test method
1) 마우스 대식세포주인 Raw 264.7을 키워 실험에 사용하였다(8 × 105 cell/well)1) Mouse macrophage cell line Raw 264.7 was used for the experiment (8 × 10 5 cells / well)
2) 12-웰 TC 플레이트에 세포를 배양한 후 1% FBS가 첨가된 DMEM에 상기에서 제조된 단삼 추출물(pH 조정)을 각각 0.1%, 0.2%, 0.5%, 1% 및 2%의 농도로 첨가하여 처리하였다.2) Cells were cultured on a 12-well TC plate and then the extracts (pH adjusted) prepared above were added to DMEM supplemented with 1% FBS at concentrations of 0.1%, 0.2%, 0.5%, 1% and 2% .
3) 음성 대조군은 1% FBS가 첨가된 DMEM만을 처리하였고, 양성 대조군은 Mouse IFN-B(500 units/ml)를 처리하였다.3) Negative control group was treated with DMEM supplemented with 1% FBS, and positive control group was treated with Mouse IFN-B (500 units / ml).
4) 단삼 처리 12시간 후 PR8-gfp(swine influenza virus)를 MOI 0.5 수준으로 접종하였다.4) PR8-gfp (swine influenza virus) was inoculated at a MOI level of 0.5 after 12 hours of treatment.
5) 접종 후 2시간 후 접종액을 제거하고, PBS로 3회 세척하고, 8~10시간 후 바이러스 감염 정도를 측정하였다.
5) After 2 hours of inoculation, the inoculum was removed, washed 3 times with PBS, and the virus infection was measured 8 ~ 10 hours later.
(2) 항바이러스 활성 실험결과(2) Results of antiviral activity test
도 5에 단삼 추출물의 농도에 따른 항바이러스 활성을 GFP 형광이미지로 나타낸다. 도 5로부터 확인할 수 있는 바와 같이, 단삼 추출물은 농도 의존적으로 양성 대조군과 유사한 정도의 항바이러스 활성을 나타내었다.
FIG. 5 shows the antiviral activity according to the concentration of the extract of Ganoderma lucidum as a GFP fluorescence image. As can be seen from FIG. 5, the extracts of Panax ginseng showed antiviral activity similar to that of the positive control in a concentration-dependent manner.
5. 마우스 5. Mouse 대식세포주를Macrophage cell line 이용한 Used 단삼의Sweet ginseng 전염증성 사이토카인 유도( Proinflammatory cytokine induction ( propro -inflammatory -inflammatory CytokineCytokine InductionInduction ) 실험) Experiment
: 단삼 추출물의 면역인자 유도효과를 확인하기 위하기 하기와 같이 실험을 수행하였다.
: Experiments were carried out as follows to confirm the induction effect of the extract of Dansamp.
(1) 전염증성 사이토카인 유도 실험방법(1) Proinflammatory cytokine induction experiment method
1) 마우스 대식세포주인 Raw 264.7을 키워 실험에 사용하였다(8 × 105 cell/well)1) Mouse macrophage cell line Raw 264.7 was used for the experiment (8 × 10 5 cells / well)
2) 6-웰 TC 플레이트에 세포를 배양한 후, 1% FBS가 첨가된 DMEM에 상기에서 제조된 단삼 추출물(pH 조정)을 1% 농도로 첨가하여 처리하였다.2) Cells were cultured on a 6-well TC plate and treated with DMEM supplemented with 1% FBS at a concentration of 1% of the extract prepared above (pH adjusted).
3) 음성 대조군은 1% FBS가 첨가된 DMEM만을 처리하였고, 양성 대조군은 리포폴리사카라이드(LPS) 분말(100ng/㎖)을 처리하였다.3) Negative control group was treated with DMEM supplemented with 1% FBS, and positive control group treated with lipopolysaccharide (LPS) powder (100ng / ml).
4) 처리 후, 6시간, 12시간, 24시간, 36시간의 결과물에 대하여 TNF-α, IL-6 및 IL-12의 값을 ELISA를 이용하여 측정하였다.4) After treatment, the values of TNF-α, IL-6 and IL-12 were measured by ELISA for the results of 6 hours, 12 hours, 24 hours and 36 hours.
(2) 전염증성 사이토카인 유도 실험결과(2) Results of induction of proinflammatory cytokine
도 6에 단삼 추출물에 의한 전염증성 사이토카인 유도 실험결과를 나타낸다(NC: 음성 대조군, LPS: 양성 대조군, 단삼: 단삼 추출물 처리군). TNF-α는 면역 및 염증반응의 매개물질로 대식세포주를 활성화시키며 다양한 사이토카인 분비를 유도하는 물질로 B-세포를 증식시키고, IL-12는 T-세포 분화를 촉진시키는 등 세포성 면역반응에 영향을 미치는 중요한 사이토카인이며, IL-6는 B-세포를 활성화시켜 항체생산을 증가시켜 항원특이적 면역반응을 촉진하는 중요한 사이토카인이다.FIG. 6 shows the results of induction of proinflammatory cytokines by the extract of Panax notoginseng (NC: negative control group, LPS: positive control group; TNF-α is an immune and inflammatory mediator that activates macrophages and induces a variety of cytokine secretion. It stimulates B-cells and IL-12 promotes T-cell differentiation, And IL-6 is an important cytokine that activates B-cells to increase antibody production and promote antigen-specific immune responses.
도 6으로부터, 단삼 추출물에 의하여 Raw cell로부터 전염증성 사이토카인 유도능이 강하게 나타나는 것을 확인할 수 있다.
From FIG. 6, it can be confirmed that the induction ability of proinflammatory cytokine from raw cells is strongly exhibited by the extract of Panax ginseng.
6. 마우스 6. Mouse 대식세포주를Macrophage cell line 이용한 Used 단삼의Sweet ginseng 인터페론-β 분비 유도 실험 Interferon-beta secretion induction experiment
: 단삼 추출물의 면역인자 유도효과를 확인하기 위하여 하기와 같이 실험을 수행하였다.
: Experiments were carried out as follows to confirm the induction effect of the extract of Radix Salviae Radix.
(1) 인터페론-β 분비 유도((1) induction of interferon-beta secretion ( InterferonInterferon -β -β secretionsecretion InductionInduction ) 실험방법) Experimental method
1) 마우스 대식세포주인 Raw 264.7을 키워 실험에 사용하였다(8 × 105 cell/well)1) Mouse macrophage cell line Raw 264.7 was used for the experiment (8 × 10 5 cells / well)
2) 6-웰 TC 플레이트에 세포를 배양한 후, 1% FBS가 첨가된 DMEM에 상기에서 제조된 단삼 추출물(pH 조정)을 1% 농도로 첨가하여 처리하였다.2) Cells were cultured on a 6-well TC plate and treated with DMEM supplemented with 1% FBS at a concentration of 1% of the extract prepared above (pH adjusted).
3) 음성 대조군은 1% FBS가 첨가된 DMEM만을 처리하였고, 양성 대조군은 리포폴리사카라이드(LPS) 분말(100ng/㎖)을 처리하였다.3) Negative control group was treated with DMEM supplemented with 1% FBS, and positive control group treated with lipopolysaccharide (LPS) powder (100ng / ml).
4) 처리 후 12시간, 24시간의 결과물에 대하여 인터페론-β의 값을 ELISA를 이용하여 측정하였다.
4) Interferon-β values were measured by ELISA for the results of 12 hours and 24 hours after the treatment.
(2) 인터페론-β 분비 유도 실험결과(2) Results of induction of interferon-beta secretion
도 7에 단삼 추출물에 의한 인터페론-β 분비 유도 실험결과를 나타낸다(NC: 음성 대조군, LPS: 양성 대조군, 단삼: 단삼 추출물 처리군). 인터페론-β는 바이러스, 암세포 등의 외부 물질에 반응하여 분비되는 사이토카인으로, 세포내 항암 및 항바이러스 작용을 일으켜 면역반응을 유도하는 물질이다.FIG. 7 shows the results of induction of interferon-beta secretion by the extract of Panax ginseng (NC: negative control group, LPS: positive control group; Interferon-beta is a cytokine secreted in response to external substances such as viruses and cancer cells, and induces an immune response by causing an anti-cancer and anti-viral action.
도 7로부터, 단삼 추출물에 의하여 Raw cell로부터 인터페론-β 유도능이 나타나는 것을 확인할 수 있다.
From FIG. 7, it can be confirmed that interferon-beta induction ability is shown from raw cells by the extract of Rawan extract.
7. 마우스에서의 7. In the mouse 단삼Sansam 추출물에 의한 인플루엔자 바이러스 감염 억제 실험 Inhibition of Influenza Virus Infection by Extracts
(1) 실험방법(1) Experimental method
1) 4-5주령의 Female Balb/c 마우스를 대조군(5마리)과 단삼 추출물 투여군(5마리)으로 각각 그룹을 나눈다.1) Divide the female Balb / c mice, 4-5 weeks old, into control group (5 animals) and ginseng extract-treated group (5 animals), respectively.
2) 단삼 투여군(5마리)의 경우 5일 동안 단삼 추출물을 약 300㎕씩 매일 경구로 투여하고, 치사량의 PR8 바이러스(10LD50)를 대조군과 단삼 추출물 투여군의 비강으로 투여하여 공격 접종을 한 후, 단삼 추출물 투여군의 경우 바이러스 공격 이후에도 단삼 추출물을 약 300㎕씩 매일 경구로 5일 동안 투여하였다.2) In the case of Dansamp group (5 animals), about 300 ㎕ of Dansamp extract was orally administered daily for 5 days, and the lethal dose of PR8 virus (10LD 50 ) was administered to the nasal passages of control group and ginseng extract group , And in the group treated with dansamp extract, about 300 μl of the extract was administered orally daily for 5 days after the virus attack.
2일마다 마우스의 체중을 측정하였고, 마우스가 치사되기까지 마우스의 생존실험을 수행하였다.
Mice were weighed every 2 days and survival experiments of mice were performed until the mice were sacrificed.
(2) 실험결과(2) Experimental results
도 8에 단삼 추출물에 의한 인플루엔자 바이러스 감염 억제 실험결과로, 마우스의 체중 변화(a) 및 생존율(b)을 나타낸다. 도 8에서와 같이, 바이러스 공격 후 대조군 마우스들이 7일 이내에 모두 치사된 것에 반해, 단삼 추출물을 투여한 그룹의 마우스 중에서는 1마리만이 치사되었고, 체중변화 역시 회복됨을 확인할 수 있다.FIG. 8 shows changes in weight (a) and survival rate (b) in mice as a result of inhibition of influenza virus infection by the extract of Panax ginseng. As shown in FIG. 8, only one mouse was killed and the weight change was recovered in the mouse of the group administered with the extract of Panax ginseng, while the control mice were all killed within 7 days after the virus attack.
이러한 결과는 단삼 추출물 내에 존재하는 면역증강성분이 마우스의 선천 면역을 증가시켜 인플루엔자 바이러스의 감염 및 증식을 억제한 것으로 고려된다.
These results suggest that the immune enhancing components in the extracts of Dansamp extract increase the innate immunity of mice and inhibit infection and proliferation of influenza virus.
8. 8. 제조예Manufacturing example
(1) 주사제(1) injections
상기 1.에서 제조된 단삼 추출물: 100 ㎎The extract of Panax ginseng prepared in the above step 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 ㎖가 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.
(2) 정제(2) tablets
상기 1.에서 제조된 단삼 추출물: 200 ㎎200 mg of the extract obtained from
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.
(3) 캡슐제(3) capsules
상기 1.에서 제조된 단삼 추출물: 100 ㎎The extract of Panax ginseng prepared in the above step 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.
(4) 환제(4) pills
상기 1.에서 제조된 단삼 추출물: 120 ㎎The dried ginseng extract prepared in the above step 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.
(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 1.에서 제조된 단삼 추출물을 적량 혼합하여, 살균함으로써 음료를 제조하였다.The extracts prepared in the
2) 기능성 식품2) Functional food
상기 1.에서 제조된 단삼 추출물을 각종 비타민 및 미네랄 함유 기능성 식품에 적량 혼합하여 단삼 추출물이 함유된 기능성 식품을 제조하였다.
The functional food containing the extract of Radix Salviae Radix was prepared by mixing the extract of Radix Salviae Radix prepared in 1 above with various vitamins and mineral-containing functional foods.
(6) 사료 조성물(6) Feed composition
동물용 배합 사료에 상기 1.에서 제조된 단삼 추출물을 적량 혼합하여, 사료 조성물을 제조한 후, 펠렛화 및 과립화하였다.
The animal feed diets were mixed with the dried fish extract prepared in the
상기 본 발명은 전술한 실시예 및 첨부된 도면에 의해 한정되는 것이 아니고, 본 발명의 기술적 사상을 벗어나지 않는 범위 내에서 여러 가지 치환, 변형 및 변경이 가능하다는 것이 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 명백할 것이다.While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments or constructions. Various changes, substitutions and alterations can be made hereto without departing from the spirit and scope of the invention. It will be clear to those who have knowledge.
Claims (13)
Immunity-enhancing and antiviral pharmaceutical composition comprising the extract of Salvia.
상기 단삼 추출물은 열수 추출, 알코올 추출 및 주정 추출로 이루어진 군으로부터 선택되는 일 이상의 방법에 의하여 형성된 것을 특징으로 하는
면역 증강 및 항바이러스용 약학 조성물.
The method of claim 1,
The Salvia extract is characterized in that formed by at least one method selected from the group consisting of hot water extraction, alcohol extraction and alcohol extraction.
A pharmaceutical composition for immune enhancement and antiviral activity.
약학적으로 허용가능한 담체, 부형제 또는 희석제를 더 포함하는 것을 특징으로 하는
면역 증강 및 항바이러스용 약학 조성물.
The method of claim 1,
It further comprises a pharmaceutically acceptable carrier, excipient or diluent
A pharmaceutical composition for immune enhancement and antiviral activity.
상기 약학적 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 현탁액, 에멀젼, 시럽제, 에어로졸, 외용제, 좌제 및 주사제로 이루어진 군으로부터 선택되는 형태인 것을 특징으로 하는
면역 증강 및 항바이러스용 약학 조성물.
The method of claim 1,
Wherein the pharmaceutical composition is in a form selected from the group consisting of tablets, pills, powders, granules, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories and injections
A pharmaceutical composition for immune enhancement and antiviral activity.
박테리아 감염성 질병 또는 바이러스 감염성 질병의 예방 또는 치료에 이용되는 것을 특징으로 하는
면역 증강 및 항바이러스용 약학 조성물.
The method of claim 1,
Used for the prevention or treatment of bacterial or viral infectious diseases
A pharmaceutical composition for immune enhancement and antiviral activity.
Health functional foods for immune enhancement and antiviral containing Salvia extract.
상기 단삼 추출물은 열수 추출, 알코올 추출 및 주정 추출로 이루어진 군으로부터 선택되는 일 이상의 방법에 의하여 형성된 것을 특징으로 하는
면역 증강 및 항바이러스용 건강기능식품.
The method according to claim 6,
The Salvia extract is characterized in that formed by at least one method selected from the group consisting of hot water extraction, alcohol extraction and alcohol extraction.
Immune enhancement and antiviral health functional food.
식품학적으로 허용가능한 식품 보조 첨가제를 더 포함하는 것을 특징으로 하는
면역 증강 및 항바이러스용 건강기능식품.
The method according to claim 6,
It further comprises a food acceptable food supplement additive
Immune enhancement and antiviral health functional food.
상기 건강기능식품은 분말, 과립, 정제, 캡슐, 캔디, 츄잉검, 젤리 및 음료로 이루어진 군으로부터 선택되는 형태인 것을 특징으로 하는
면역 증강 및 항바이러스용 건강기능식품.
The method according to claim 6,
Wherein the health functional food is a form selected from the group consisting of powder, granule, tablet, capsule, candy, chewing gum, jelly and beverage
Immune enhancement and antiviral health functional food.
Immunity-enhancing and antiviral feed composition comprising the extract of Salvia.
상기 단삼 추출물은 열수 추출, 알코올 추출 및 주정 추출로 이루어진 군으로부터 선택되는 일 이상의 방법에 의하여 형성된 것을 특징으로 하는
면역 증강 및 항바이러스용 사료 조성물.
11. The method of claim 10,
The Salvia extract is characterized in that formed by at least one method selected from the group consisting of hot water extraction, alcohol extraction and alcohol extraction.
Feed composition for immune enhancing and antiviral.
A method for enhancing immunity and antiviral activity of an animal comprising administering to the animal in need of immune boosting, extract of Salvia soybean.
상기 단삼 추출물은 열수 추출, 알코올 추출 및 주정 추출로 이루어진 군으로부터 선택되는 일 이상의 방법에 의하여 형성된 것을 특징으로 하는
동물의 면역 증강 및 항바이러스 활성 증강 방법.The method of claim 12,
The Salvia extract is characterized in that formed by at least one method selected from the group consisting of hot water extraction, alcohol extraction and alcohol extraction.
Methods of Enhancing Immunity and Antiviral Activity in Animals.
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