KR101400893B1 - A composition for the enhancement of immune system comprising extracts or fractions of eremochloa ophiuroides as an active ingredient - Google Patents
A composition for the enhancement of immune system comprising extracts or fractions of eremochloa ophiuroides as an active ingredient Download PDFInfo
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- KR101400893B1 KR101400893B1 KR1020120132290A KR20120132290A KR101400893B1 KR 101400893 B1 KR101400893 B1 KR 101400893B1 KR 1020120132290 A KR1020120132290 A KR 1020120132290A KR 20120132290 A KR20120132290 A KR 20120132290A KR 101400893 B1 KR101400893 B1 KR 101400893B1
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- extract
- fraction
- active ingredient
- centipede
- ethyl acetate
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/324—Foods, ingredients or supplements having a functional effect on health having an effect on the immune system
Abstract
Description
본 발명은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물 또는 이의 분획물을 유효성분으로 함유하는 면역 증강용 조성물에 관한 것이다.
The present invention relates to centipede grass ( Eremochloa ophiuroides extract or fractions thereof as an active ingredient.
면역은 크게, 태어날 때부터 지니고 있는 선천면역(innate immunity)과 후천적으로 생활 등에 적응되어 얻어지는 획득면역(acquired immunity)으로 구분된다. 선천면역은 일명 '자연 면역'이라고도 하며 항원에 대해 비특이적으로 반응하며 특별한 기억작용은 없다. 선천적인 면역체계로는 항원의 침입을 차단하는 피부, 점액조직, 상산성의 위산, 혈액에 존재하는 보체(complement) 등이 있다. 세포로는 식균작용을 담당하는 대식세포(macrophage)와 다형핵 백혈구(polymorphonuclear leukocyte), 감염세포를 죽일 수 있는 자연 살해(NK)세포 등이 있다. 실제로 대부분의 감염은 이 선천면역에 의해 방어된다. 또한, 획득면역은 일명 '후천 면역'이라고도 하며 처음 침입한 항원에 대해 기억할 수 있고 다시 침입할 때 특이적으로 반응하여 효과적으로 항원을 제거할 수 있는 특징이 있어 선천 면역을 보강하는 역할을 한다. 흔히 사용되는 면역의 정의는 이것을 말한다. 이 획득면역은 편의상 체액성 면역(humoral immunity)과 세포성 면역(cell-mediated immunity)으로 나누어 이해된다. 체액성 면역은 B 림프구가 항원을 인지한 후 분화되어 항체를 분비하고, 이 항체는 주로 감염된 세균을 제거하는 기능을 보여준다. 항체는 체액에 존재하며 면역글로불린(immunoglobulin: Ig)이라는 당단백질로 이루어져 있다. 여기에는 IgG, IgM, IgA, IgD, IgE 등이 있으며, 각각 독특한 기능을 수행하며 일부 기능이 중복되기도 한다. IgA 항체는 태반을 통해 태아에 전달되는 특징이 있다. 이와 같은 면역을 모성 면역(maternal immunity)이라 하며, 이 때문에 출생 후 수개월 동안 잘 감염되지 않는다. 세포성 면역은 흉선에서 유래한 T림프구가 항원을 인지하여 림포카인(lymphokine)을 분비하거나 직접 감염된 세포를 죽이는 역할을 한다. 분비된 림포카인은 대식세포를 활성화시켜 식작용(phagocytosis)을 돕기도 한다. 이와 같은 세포성 면역은 주로 바이러스 또는 세포내에서 자랄 수 있는 세균에 감염된 세포를 제거하는 기능을 수행한다. 획득면역은 병원체 또는 그 독소를 면역원으로 예방접종하여 얻을 수 있으며, 이와 같은 면역을 인공면역(artificial immunity)이라 한다. 면역기능이 저하되면, 과민반응(allergy, hypersensitivity)을 일으키는 경우가 있는데 알러지와 같은 과민반응은 발현시간과 반응형태에 기초하여 네가지 타입으로 나눌 수 있다.Immunity is largely divided into innate immunity, which is born from birth, and acquired immunity, which is obtained by adaptation to life and the like. Congenital immunity is also called 'natural immunity' and responds nonspecifically to antigens and has no special memory effect. Congenital immune systems include skin, mucus tissue, gastric acid that blocks antigen entry, and complement in blood. Cells include macrophages, polymorphonuclear leukocytes, and natural killer (NK) cells that can kill infected cells. In fact, most infections are defended by this congenital immune system. Acquired immunity is also known as 'acquired immune', and it can remember about the first invaded antigen, reacts specifically when invading again, and can effectively remove the antigen, thereby reinforcing innate immunity. The commonly used definition of immunity is this. This acquisition immunity is understood to be divided into humoral immunity and cell-mediated immunity for convenience. Humoral immunity recognizes B-lymphocytes and then differentiates and secretes antibodies. This antibody mainly functions to remove infected bacteria. The antibody is present in body fluids and consists of a glycoprotein called immunoglobulin (Ig). These include IgG, IgM, IgA, IgD, and IgE, each of which performs a unique function, and some functions may overlap. The IgA antibody is characterized by its transfer to the fetus through the placenta. This type of immunity is called maternal immunity, and it is not well infected for several months after birth. Cell-mediated immunity recognizes antigens from T-lymphocytes derived from the thymus and secretes lymphokines or kills directly infected cells. Secreted lymphocytes also activate phagocytosis by activating macrophages. Such cell-mediated immunity mainly functions to remove cells infected with a virus or bacteria that can grow in the cell. Acquired immunity can be obtained by immunizing a pathogen or its toxin with an immunogen, and such immunity is called artificial immunity. When immune function deteriorates, it causes allergy (hypersensitivity). Hypersensitivity such as allergies can be divided into four types based on the time of expression and the type of reaction.
제I형(즉시형)은 대부분의 알러지 반응들이 속하는 전형적인 것으로서, 그 증상은 천식, 계절성 혹은 통년성 비염, 알러지성 부비강염, 결막염, 식품과 약물에 의한 알러지, 아토피성 피부염, 두드러기, 벌침에 대한 알러지 반응 등이 있다. 제Ⅱ형(항체관여)은 체액성 면역의 세포용해 반응이나 염증반응을 일으키는데 증상으로는 부적합 수혈에 의한 적혈구의 용혈, 신생아 용혈성 질환이 있다. 제Ⅲ형은 항원-항체 결합에 의한 반응이나 작은 분자량의 항원에 의해 발생한다. 급성 사구체 신염, 과민성 폐렴을 일으킨다. 마지막으로, 제Ⅳ형(지연형)은 T 임파구와 대식세포에 의한 세포성 면역 반응의 한 형태로 결핵균의 투베르쿨린반응, 이식거부반응, 바이러스와 같은 미생물이 침입에 대한 반응 등의 중요한 반응을 일으킨다.Type I (immediate type) is typical of most allergic reactions, and its symptoms are allergic to seasonal or perennial rhinitis, allergic sinusitis, conjunctivitis, allergies due to food and drugs, atopic dermatitis, Reaction and the like. Type II (antibody involvement) causes cytolytic or inflammatory reactions of humoral immunity. Symptoms include red blood cell hemolysis due to an inadequate transfusion, and hemolytic disease of the newborn. Type III is caused by antigen-antibody binding or by small molecular weight antigens. Acute glomerulonephritis, hypersensitivity pneumonia. Finally, Type IV (delayed type) is a form of cellular immune response by T lymphocytes and macrophages, leading to important reactions such as tuberculin reaction of tubercle bacilli, transplant rejection, and response to microbes such as viruses .
상술한 바와 같은 원인을 이유로 하여, 면역기능에 장애가 발생하는 문제점을 극복하기 위하여, 다양한 면역기능 증강제 및 치료제가 개발되어 사용되고 있다. 그러나, 별도의 치료제를 장기간 복용할 경우, 개개인마다 면역력의 차이가 달라서 그 부작용의 문제점이 크다는 단점이 있다. 특히, 면역 질환은 치료보다 예방이 중요한데, 현재 시판되고 있는 치료제는 예방을 위하여 복용하기 어려운 것이 현실이다. 따라서 부작용의 가능성이 있는 별도의 면역기능 강화제를 복용하지 않고서도, 면역기능 장애를 예방 및 치료할 수 있는 방법이 요구된다.Various immune function-enhancing agents and therapeutic agents have been developed and used in order to overcome the problem that the immune function is impaired due to the above-described causes. However, when a separate therapeutic agent is taken for a long period of time, there is a disadvantage in that the immunity of each individual is different and the problems of the side effects are large. In particular, prevention of immune diseases is more important than treatment, and it is a reality that currently available therapeutic agents are difficult to take for prevention. Therefore, there is a need for a method that can prevent and treat immune dysfunction without taking additional immunity enhancers that may have side effects.
일반적으로 지금까지 알려진 면역증강 물질로는 크게 버섯 추출물과 식물 추출물이 많이 있으며 이러한 소재의 효과에 대한 보고가 계속되고 있다. 버섯추출물로는 크레스틴, 렌티난 및 AHCC가 대표적이며 식물추출물로는 미슬토가 대표적인 면역력을 증강시키는 물질이다.
In general, there are many mushroom extracts and plant extracts as immunopotentiating substances so far known. Mushroom extracts include crestin, lentinan, and AHCC. Plant extracts are a typical example of mushrooms that enhance immunity.
최근 국민 소득과 경제 수준 향상으로 건강에 대한 관심이 고조되면서 축산물의 양적 측면 보다 질적 측면이 강조되고 있으며, 축산물의 품질 향상 및 안정성 확보는 국내 축산업의 경쟁력 확보는 물론 국민 건강을 위해 더욱 중요시되고 있다. 가축의 생산성 극대화 및 질병 예방을 목적으로 성장촉진용 항생제가 사용되어 왔으나, 이는 소비자에게 부정적으로 인식되고 있으며 축산물 내 항생제 잔류, 내성균 출현은 물론 생태계 파괴 등의 문제를 유발시킬 수 있다. 이에 따라 국내에서는 2004년부터 항생제 등 항균 물질 사용 절감 방안이 추진되어 2012년에는 배합사료 내 성장촉진용 항생제의 사용이 전면 금지될 예정이다. 또한 무항생제 및 친환경 축산물 인증제가 추진되면서 무항생제 사육에 대한 관심 역시 높아지고 있으며 일반 축산물과의 품질 차별화가 가속화되고 있다. 그러나 무항생제 사육시 생산 비용 증가, 가축 생산성 감소, 질병 발생율 및 폐사율 증가 등의 문제가 발생되어 그 대응책이 시급한 실정이다. 현재 유기산, 생균제, prebiotics, 식물추출물 등 다양한 종류의 항생제 대체제가 개발, 사용되고 있으나 이용효과, 경제성 등 다양한 측면에서 항생제 대비 효율성이 낮고 여러 스트레스 요인이 산재하는 농장 단위에서 현재 시판, 사용되고 있는 항생제 대체제를 통한 효과적인 대체는 현실적으로 어렵기 때문에 성장촉진용 항생제 대체 및 스트레스 저감 효과가 우수한 생리 활성물질을 개발하기 위한 연구가 활발히 진행되고 있다.
In recent years, interest in health has been heightened by the improvement of national income and economic level, and the qualitative aspect of livestock products has been emphasized. Quality improvement and stability of livestock products have become more important for national health as well as competitiveness of domestic livestock industry . Antibiotics for growth promotion have been used for the purpose of maximizing the productivity of livestock and preventing disease. However, this has been recognized as a negative for consumers, and it can cause problems such as antibiotic residues in livestock products, emergence of resistant bacteria and destruction of ecosystem. As a result, domestic use of antibiotics such as antibiotics will be reduced in 2004, and the use of antibiotics for promoting growth in compound feeds will be prohibited in 2012. In addition, as non-antibiotic and eco-friendly livestock products certification system is promoted, interest in non-antibiotic production is increasing, and quality differentiation from general livestock products is accelerating. However, in the case of non-antibiotics, problems such as increase of production cost, decrease of livestock productivity, incidence of disease and increase of mortality rate are urgently needed. Currently, a variety of antibiotic substitutes such as organic acids, probiotics, prebiotics, and plant extracts have been developed and used. However, antibiotic substitutes are currently being marketed and used in farms with low efficiency compared to antibiotics and various stress factors. , It has been actively studied to develop physiologically active substances having excellent antibiotic substitution and stress reduction effect for growth promotion.
센티페드그라스(centipede grass, Eremochloa ophiuroides)는 생육이 늦은 난지형 잔디로서 뗏장을 형성하며 포복 생장을 하는 다년생 잔디이다. 벼과 식물에 속하며, 학명은 Eremochloa ophiuroides (Munro) Hack이다. 주로 중국과 동아시아, 인도차이나, 미국 북동부, 메소아메리카와 캐러비안 지역에 분포하고 있으며, 각종 토양조건에 폭 넓게 잘 적응되지만, 특히 습하며 산성토양 및 비옥도가 낮은 사질 토양에서 잘 견딘다.Centipede grass ( Eremochloa ophiuroides ) is a perennial turf that grows slowly and forms a turf as a turfgrass. It belongs to the genus Porphyra, and its scientific name is Eremochloa ophiuroides (Munro) Hack. It is mainly distributed in China, East Asia, Indochina, Northeastern USA, Mesoamerica and the Caribbean. It is well adapted to various soil conditions, but it is particularly resistant to moist and acid soil and low fertility soil.
형태학적 특징으로서, 잎은 길이 15~30 ㎜, 폭 2~4 ㎜이고, 흰 중심맥(mid vein)을 가지는 납작한 모양이며, 경령(collar) 부분을 제외하고 털이 없고, 잎 끝부분은 둥글다. 줄기는 아주 굳게 다문 잎집(sheath)을 가지고, 뿌리는 가는 분기형의 포복경(stolons)이다. 꽃 부분은 3~5 인치 정도의 하나의 총상화서(總狀花序, raceme, = 총상꽃차례)를 가지고, 상기 총상화서는 자줏빛색을 띠고 약간 평평하며 두 줄로 배열된 작은 이삭(小穗, spikelet)을 가진다. 주용도는 잔디 밭 또는 뗏장 잔디로 사용되고 높은 잎의 비율(leafiness)과 좋은 맛 때문에 조사료로 이용된다.
As a morphological feature, the leaf has a length of 15 ~ 30 ㎜, a width of 2 ~ 4 ㎜, a flat shape with a mid vein, no hairs except a collar part, and a leaf tip round. The stem is a branching type stolons with a very firmly sheath (sheath). The flower part has a three to five inches long fossil flower (raceme), which has a purple color and a slightly flat, spikelet arranged in two lines. I have. It is used as a lawn or turf grass and is used as a forage for its leafiness and good taste.
이에 본 발명자들은 면역 증강 효과를 갖는 천연 식물 추출물을 연구하던 중, 센티페드그라스에서 추출한 추출물, 이의 분획물 및 상기 분획물로부터 분리한 활성분획이 면역 반응과 관련된 사이토카인(IL-6, IL-10) 및 STAT3의 인산화를 억제함으로써 면역 증강용 조성물의 유효성분으로 유용하게 사용될 수 있음을 밝힘으로써 본 발명을 완성하였다.
Accordingly, the present inventors have found that when extracting natural plant extracts having an immunostimulating effect, extracts from Centipede grass, fractions thereof, and active fractions isolated from the fractions are useful as cytokines (IL-6, IL-10) And STAT3 phosphorylation, and thus the present invention has been completed.
본 발명의 목적은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물 또는 이의 분획물을 유효성분으로 함유하는 면역 증강용 조성물을 제공하는 것이다.
The object of the present invention is to provide a centipede grass ( Eremochloa ophiuroides extract or a fraction thereof as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 센티페드그라스스(centipede grass, Eremochloa ophiuroides) 추출물을 유효성분으로 함유하는 면역 증강용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for enhancing immunity, comprising centipede grass ( Eremochloa ophiuroides) extract as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출하여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for enhancing immunity, which comprises an organic solvent fraction prepared by further extracting a centipedragus extract with an organic solvent as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 유효성분으로 함유하는 면역 증강용 건강식품을 제공한다.In addition, the present invention provides a health food for immuno-enhancement comprising a centipedragus extract as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출하여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 건강식품을 제공한다. The present invention also provides a health food for immunity enhancement comprising an organic solvent fraction prepared by further extracting a centipedragus extract with an organic solvent as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 유효성분으로 함유하는 면역 증강용 사료 조성물을 제공한다.In addition, the present invention provides an immunostimulating feed composition comprising a centipede glass extract as an active ingredient.
아울러, 본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출하여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 사료 조성물을 제공한다.
In addition, the present invention provides an immunomodulating dietary composition comprising an organic solvent fraction prepared by further extracting a centiped glass extract with an organic solvent as an active ingredient.
본 발명은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물, 이의 분획물 및 상기 분획물로부터 분리한 활성분획이 면역 반응과 관련된 사이토카인(IL-6, IL-10) 및 STAT3의 인산화를 억제함으로써, 면역 증강용 조성물의 유효성분으로 유용하게 사용할 수 있다.
The present invention relates to centipede grass ( Eremochloa ophiuroides extract, its fractions and the active fractions isolated from the fractions inhibit the phosphorylation of cytokines (IL-6, IL-10) and STAT3 associated with the immune response, thereby being usefully used as an active ingredient of immunomodulating compositions have.
도 1은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물, 이의 분획물 또는 상기 분획물로부터 분리한 활성 분획을 제조하는 과정을 나타낸 도이다.
도 2는 대식세포 Raw 264.7 세포에 지다당류(lipopolysaccharide; LPS)에 의해 유도된 면역 반응에 대한 센티페드그라스 분획물이 사이토카인(인터루킨-6(interleukin-6, IL-6), 인터루킨-10(IL-10))에 미치는 영향을 보기 위해 효소 면역 측정법을 통해 확인한 도이다.
도 3은 7주령 ICR 마우스에서 LPS에 의해 유도된 면역 반응에 대한 센티페드그라스 분획물이 사이토카인(IL-6, IL-10)에 미치는 영향을 보기 위해 효소 면역 측정법을 통해 확인한 도이다.
도 4는 대식세포 Raw 264.7 세포에 LPD에 의해 유도된 면역에 관련된 단백질들의 변화를 웨스터블롯을 통해 확인한 도이다. Figure 1 is a schematic view of a centipede grass ( Eremochloa ophiuroides extract, fractions thereof, or active fractions isolated from the fractions.
Figure 2 shows that the centipede grass fraction for the immune response induced by lipopolysaccharide (LPS) on macrophage Raw 264.7 cells is a cytokine (interleukin-6, IL-6), interleukin-10 -10)) in order to examine the effect of the enzyme immunoassay method.
FIG. 3 is a graph showing the effect of LPS-induced immune responses on 7-week-old ICR mice on cytokines (IL-6, IL-10) by enzyme immunoassay.
FIG. 4 is a graph showing changes in proteins related to immunity induced by LPD in macrophage Raw 264.7 cells through Western blotting.
이하, 본 발명을 상세히 설명한다.
Hereinafter, the present invention will be described in detail.
본 발명은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물을 유효성분으로 함유하는 면역 증강용 약학적 조성물 제공한다.The present invention relates to centipede grass ( Eremochloa ophiuroides ) as an active ingredient.
상기 센티페드그라스 추출물은 하기의 단계들을 포함하는 제조방법에 의해 제조되는 것이 바람직하나 이에 한정하지 않는다:The centipedlass extract is preferably, but not exclusively, prepared by a process comprising the steps of:
1) 센티페드그라스에 추출용매를 가하여 추출하는 단계;1) extracting Sentipedgrass with an extraction solvent;
2) 단계 1)의 추출물을 식힌 후 여과하는 단계; 및2) cooling the extract of step 1) and filtering; And
3) 단계 2)의 여과한 추출물을 감압농축하는 단계.3) Concentrating the filtered extract of step 2) under reduced pressure.
상기 방법에 있어서, 단계 1)의 센티페드그라스 재배한 것 또는 시판되는 것 등 제한 없이 사용할 수 있다. 상기 센티페드그라스는 잎, 줄기 또는 뿌리를 모두 이용할 수 있으나, 잎을 이용하는 것이 바람직하다.In the above method, cultivated Centipede grains of step 1) or commercially available ones can be used without limitation. The centipedegrass may be a leaf, a stem, or a root, but leaves are preferably used.
상기 제조방법에 있어서, 단계 1)의 추출용매는 물, 알코올 또는 이들의 혼합물인 것이 바람직하고, 상기 알코올로는 C1 내지 C2 저급 알코올을 이용하는 것이 바람직하며, 저급 알코올로는 에탄올 또는 메탄올을 이용하는 것이 바람직하고, 메탄올을 이용하는 것이 더욱 바람직하나 이에 한정되지 않는다. 추출방법으로는 진탕추출 또는 환류추출을 이용하는 것이 바람직하나 이에 한정하지 않는다. 상기 추출용매를 건조된 센티페드그라스의 총 분량에 2 내지 10배 첨가하여 추출하는 것이 바람직하며, 10배 첨가하여 추출하는 것이 더욱 바람직하나 이에 한정되지 않는다. 또한, 추출온도는 50 내지 120℃인 것이 바람직하며, 40 내지 45℃인 것이 더욱 바람직하나 이에 한정하지 않는다. 또한, 추출시간은 2 내지 5일인 것이 바람직하나 이에 한정하지 않는다. 아울러, 추출 회수는 1 내지 3회인 것이 바람직하나 이에 한정되는 것은 아니다.In the above production method, the extraction solvent of step 1) is preferably water, alcohol or a mixture thereof, and the alcohol is preferably C 1 To C 2 lower alcohol is preferably used. As the lower alcohol, ethanol or methanol is preferably used, and methanol is more preferably used, but not limited thereto. As the extraction method, it is preferable to use shaking extraction or reflux extraction, but it is not limited thereto. The extraction solvent is preferably added by 2 to 10 times the total amount of the dried centipedgrass, more preferably by 10 times, but not limited thereto. Further, the extraction temperature is preferably 50 to 120 ° C, more preferably 40 to 45 ° C, but is not limited thereto. The extraction time is preferably 2 to 5 days, but is not limited thereto. In addition, the number of times of extraction is preferably 1 to 3 times, but is not limited thereto.
상기 방법에 있어서, 단계 3)의 감압농축은 진공감압농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 상기 감압농축은 20 내지 60℃에서 농축하는 것이 바람직하고 45 내지 55℃에서 농축하는 것이 바람직하나 이에 한정되지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무건조 또는 동결건조하는 것이 바람직하나 이에 한정하지 않는다. 상기 동결건조는 -50 내지 -100℃에서 건조하는 것이 바람직하고 -70℃에서 건조하는 것이 바람직하나 이에 한정되지 않는다.In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The concentration under reduced pressure is preferably 20 to 60 占 폚, and is preferably concentrated at 45 to 55 占 폚, but is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto. The lyophilization is preferably performed at -50 to -100 캜, but drying is preferably performed at -70 캜, but is not limited thereto.
본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출하여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for immunostaining comprising an organic solvent fraction prepared by further extracting a centipedragus extract with an organic solvent as an active ingredient.
본 발명은 센티페드그라스 추출물의 분획물을 추가적으로 컬럼크로마토그래피로 분리한 활성분획을 유효성분으로 함유하는 면역 증강용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for immunostaining comprising an active fraction obtained by further fractionating a fraction of Sentifedgrax extract by column chromatography as an active ingredient.
상기 센티페드그라스 추출물의 분획물 또는 상기 분획물로부터 분리한 활성분획은 하기의 단계들을 포함하는 제조방법에 의해 제조되는 것이 바람직하나 이에 한정하지 않는다:The fraction of the centipedlass extract or the active fraction isolated from the fraction is preferably, but not necessarily, prepared by a process comprising the steps of:
1) 센티페드그라스 추출물에 추가로 유기용매를 가하여 유기용매 분획물을 제조하는 단계; 및1) preparing an organic solvent fraction by adding an organic solvent to the centipede glass extract; And
2) 단계 1)의 분획물을 컬럼크로마토그래피를 실시하여 활성분획을 제조하는 단계.2) Performing column chromatography on the fraction of step 1) to produce an active fraction.
상기 방법에 있어서, 단계 1)의 유기용매는 n-헥산 또는 에틸아세테이트인 것이 바람직하나 이에 한정하지 않는다.In the above method, the organic solvent in step 1) is preferably, but not limited to, n-hexane or ethyl acetate.
상기 방법에 있어서, 상기 유기용매 분획물은 n-헥산으로 추출한 후 물층을 에틸아세테이트로 추출한 에틸아세테이트 분획물인 것이 바람직하나 이에 한정하지 않는다.In the above method, the organic solvent fraction is preferably an ethylacetate fraction extracted with n-hexane and then a water layer extracted with ethyl acetate, but not always limited thereto.
상기 방법에 있어서, 단계 2)의 컬럼크로마토그래피는 실리카겔, 세파덱스, RP-18, 폴리아미드, 도요펄(Toyopearl) 및 XAD 수지로 이루어진 군으로부터 선택된 충진제를 이용한 컬럼크로마토그래피를 수행하여 활성분획을 분리 및 정제할 수 있다. 컬럼크로마토그래피는 필요에 따라 적절한 충진제를 선택하여 수차례 실시할 수 있다.In this method, the column chromatography in step 2) is carried out by column chromatography using a filler selected from the group consisting of silica gel, Sephadex, RP-18, polyamide, Toyopearl and XAD resin to obtain an active fraction Can be separated and purified. The column chromatography can be carried out several times by selecting an appropriate filler as necessary.
본 발명자들은 센티페드그라스의 에틸아세테이트 분획물을 Flex-ColumTM(3 cm × 25 cm) 및 도요펄(Toyopearl) HW-40(75 ㎛) 컬럼크로마토그래피에 흡착시키고 20, 50 또는 70% 에탄올 용출용매를 사용하여, 에틸아세테이트-20%메탄올, 에틸아세테이트-50%메탄올 및 에틸아세테이트-70%메탄올 활성분획을 얻었다.
We have found that the ethylacetate fractions of Sentifedgrass are adsorbed on Flex-Colum ™ (3 cm × 25 cm) and Toyopearl HW-40 (75 μm) column chromatography and eluted in 20, 50 or 70% Ethyl acetate-20% methanol, ethyl acetate-50% methanol and ethyl acetate-70% methanol active fractions were obtained.
본 발명의 구체적인 실시예에서, 본 발명자들은 센티페드그라스 추출물 또는 이의 분획물의 면역 증강 효과를 확인하기 위하여, 대식세포 Raw 264.7 세포에 LPS에 의해 유도된 면역 반응에 센티페드그라스 분획물을 처리한 결과, 사이토카인(IL-6, IL-10) 분비정도가 현저히 저해되는 것을 확인하였다(도 2 참조).In a specific example of the present invention, the inventors of the present invention have found that treatment of the centipede grass fraction with LPS-induced immune response to macrophage Raw 264.7 cells, in order to confirm the immunopotentiating effect of the centipedragus extract or its fraction, It was confirmed that the degree of secretion of cytokines (IL-6, IL-10) was remarkably inhibited (see Fig. 2).
또한, 본 발명자들은 센티페드그라스 추출물의 면역 증강 효과를 확인하기 위하여, ICR 마우스에 LPS를 복강투여전 2주간 센티페드그라스 추출물이 함유된 사료를 먹인 결과, LPS의 복강투여로 인한 사이토카인(IL-6, IL-10)의 증가를 50% 수준으로 억제하는 것을 확인하였다(도 3 참조).In order to confirm the immunopotentiating effect of the centipedragus extract, the present inventors administered LPS to the ICR mouse for 2 weeks before the administration of the peritoneal administration, and found that the LPS-induced cytokine IL -6, and IL-10) at the level of 50% (see FIG. 3).
또한, 센티페드그라스 70% 메탄올 분획물의 면역 관련 단백질을 확인한 결과, IL-6 발현에 깊이 관여하는 단백질인 STAT3의 인산화가 센티페드그라스 70% 메탄올 분획물 20 ㎍/㎖ 농도이상으로 처리한 군에서 현저하게 감소하였고, COX-2 단백질은 100 ㎍/㎖으로 처리한 군에서만 발현이 감소되는 것을 확인하였다(도 4 참조).In addition, immunocompetent proteins of the 70% methanol fraction of centipedegrass were identified. As a result, phosphorylation of STAT3, a protein deeply involved in the expression of IL-6, was remarkably observed in the group treated with a concentration of more than 20 μg / ml of the 70% methanol fraction of centipedegrass , And the expression of COX-2 protein was reduced only in the group treated with 100 μg / ml (see FIG. 4).
따라서, 센티페드그라스 추출물, 이의 분획물 및 상기 분획물로부터 분리한 활성분획이 면역 반응과 관련된 사이토카인(IL-6, IL-10) 및 STAT3의 인산화를 억제함으로써, 면역 증강용 조성물의 유효성분으로 유용하게 사용할 수 있다.
Accordingly, the centipede-grass extract, its fractions, and the active fractions isolated from the fractions inhibit the phosphorylation of cytokines (IL-6, IL-10) and STAT3 associated with the immune response, Can be used.
본 발명의 약학적 조성물은 통상적으로 부형제, 붕해제, 감미제, 활택제, 향미제 등을 추가로 포함할 수 있으며, 통상적인 방법에 의해 정제, 캅셀제, 산제, 과립제, 현탁제, 유제, 시럽제, 기타 액제로 제형화될 수 있다.The pharmaceutical composition of the present invention may further contain an excipient, a disintegrant, a sweetening agent, a lubricant, a flavoring agent and the like, and may be formulated into tablets, capsules, powders, granules, suspensions, emulsions, syrups, Other liquid formulations may be formulated.
구체적으로 본 발명의 약학적 조성물은 경구 투여용 제형, 예를 들면 정체, 트로치제(troches), 로젠지(lozenge), 수용성 또는 우성현탁액, 조제분말 또는 과립, 에멀젼, 하드 또는 소프트 캡슐, 시럽 또는 엘릭시르제(elixirs)로 제제화된다. 정제 및 캡슐 등의 제형으로 제제하기 위해 락토오스, 사카로오스, 솔비톨, 만니톨, 전분, 아밀로펙틴, 셀룰로오스 또는 젤라틴과 같은 결합제, 디칼슘 포스페이트와 같은 부형제, 옥수수 전분 또는 고구마 전분과 같은 붕해제, 스테아르산 마르네슘, 스테아르산 칼슘, 스테아릴푸마르산 나트륨 또는 폴리에틸렌글리콜 왁스와 같은 윤활유가 함유된다. 캡슐제형의 경우는 상기에서 언급한 물질 이외에도 지방유와 같은 액체 담체를 함유한다.Specifically, the pharmaceutical compositions of the present invention can be formulated for oral administration, for example, as tablets, troches, lozenges, aqueous or aqueous suspensions, prepared powders or granules, emulsions, hard or soft capsules, It is formulated into elixirs. Binders such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose or gelatin, excipients such as dicalcium phosphate, disintegrants such as corn starch or sweet potato starch, disintegrants such as starch, , Calcium stearate, sodium stearyl fumarate, or polyethylene glycol wax. In the case of a capsule formulation, in addition to the above-mentioned substances, a liquid carrier such as fatty oil is contained.
또한, 본 발명의 약학적 조성물은 경구 또는 비경구 투여할 수 있으며, 비경구 투여시 피하주사, 정맥주사, 근육내 주사 또는 흉부내 주사 주입방식을 선택하는 것이 바람직하다. 비경구 투여용 제형으로 제제화하기 위해서는 본 발명의 센티페드그라스 추출물, 이의 분획물 또는 분획물의 활성분획을 안정제 또는 완충제와 함께 물에서 혼합하여 현탁액으로 제조하고 이를 앰플 또는 바이알의 단위 투여형으로 제제한다.In addition, the pharmaceutical composition of the present invention can be administered orally or parenterally, and it is preferable to select subcutaneous injection, intravenous injection, intramuscular injection, or intra-thoracic injection injection method for parenteral administration. In order to formulate the formulation for parenteral administration, the active fraction of the centipede glass extract, fraction or fraction thereof of the present invention is mixed with water with a stabilizer or a buffer to prepare a suspension, which is formulated into a unit dosage form of ampoule or vial.
본 발명에 따른 유효성분의 투여량은 체내에서 활성성분의 흡수도, 불활성화율 및 배설속도, 환자의 연령, 성별 및 상태, 치료할 질병의 중증 정도에 따라 적절히 선택되나, 경구 투여제의 경우 일반적으로 성인에게 1일에 체중 1 kg당 본 발명의 추출물을 0.0001 ~ 500 mg의 양으로 1회 내지 수회 나누어 투여할 수 있으며, 0.001 ~ 100 mg의 양으로 투여하는 것이 바람직하다.
The dosage of the active ingredient according to the present invention is appropriately selected depending on the degree of absorption, inactivation rate and excretion rate of the active ingredient in the body, the age, sex and condition of the patient, and severity of the disease to be treated. The extract of the present invention may be administered to an adult in an amount of 0.0001-500 mg per 1 kg of body weight per day, preferably in an amount of 0.001-100 mg.
본 발명은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물을 유효성분으로 함유하는 면역 증강용 건강식품을 제공한다.The present invention relates to centipede grass ( Eremochloa ophiuroides ) extract as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출하여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 건강식품을 제공한다.The present invention also provides a health food for immunity enhancement comprising an organic solvent fraction prepared by further extracting a centipedragus extract with an organic solvent as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물의 분획물을 추가적으로 컬럼크로마토그래피로 분리한 활성분획을 유효성분으로 함유하는 면역 증강용 건강식품을 제공한다.The present invention also provides a health food for immunoenhancing comprising an active fraction obtained by further fractionating a fraction of a centipedrax extract by column chromatography as an active ingredient.
본 발명의 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물, 이의 분획물 및 상기 분획물로부터 분리한 활성분획이 면역 반응과 관련된 사이토카인(IL-6, IL-10) 및 STAT3의 인산화를 억제함으로써, 면역 증강용 건강식품의 유효성분으로 유용하게 사용할 수 있다.
The centipede grass of the present invention ( Eremochloa ophiuroides extract, fractions thereof and active fractions isolated from the fractions inhibit the phosphorylation of cytokines (IL-6, IL-10) and STAT3 associated with the immune response, thereby being usefully used as an active ingredient of an immunomodulating health food .
본 발명의 건강식품은 센티페드그라스 추출물, 이의 분획물 또는 상기 분획물로부터 분리한 활성분획을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다.The health food of the present invention may be added to the centipede glass extract, its fraction or the active fraction isolated from the fraction as it is, or may be used together with other food or food ingredients, and may be suitably used according to a conventional method.
상기 식품의 종류에는 특별한 제한은 없다. 상기 센티페드그라스 추출물, 이의 분획물 또는 상기 분획물로부터 분리한 활성분획을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙동제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of the food. Examples of the food to which the centipedegrass extract, the fraction thereof, or the active fraction isolated from the fraction can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gum, ice cream And various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, all of which include health foods in a conventional sense.
본 발명은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물을 유효성분으로 함유하는 면역 증강용 동물 사료 첨가제를 제공한다.The present invention provides an animal feed additive for immunity enhancement containing centipede grass (Eremochloa ophiuroides) extract as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출?여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 동물 사료 첨가제를 제공한다.In addition, the present invention provides an animal feed additive for immunity enhancement comprising an organic solvent fraction prepared by further extracting a centipede grass extract with an organic solvent as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물의 분획물을 추가적으로 컬럼크로마토그래피로 분리한 활성분획을 유효성분으로 함유하는 면역 증강용 동물 사료 첨가제를 제공한다.The present invention also provides an animal feed additive for immunity enhancement, comprising an active fraction obtained by further fractionating the fraction of Sentifed glass extract by column chromatography, as an active ingredient.
본 발명의 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물, 이의 분획물 및 상기 분획물로부터 분리한 활성분획이 면역 반응과 관련된 사이토카인(IL-6, IL-10) 및 STAT3의 인산화를 억제함으로써, 면역 증강용 동물 사료 첨가제의 유효성분으로 유용하게 사용할 수 있다.
The centipede grass of the present invention ( Eremochloa ophiuroides extract, its fractions and active fractions isolated from the fractions inhibit the phosphorylation of cytokines (IL-6, IL-10) and STAT3 associated with the immune response, thereby being useful as an active ingredient of animal feed additives for immunity enhancement Can be used.
본 발명은 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물을 유효성분으로 함유하는 면역 증강용 가축 사료 첨가제를 제공한다.The present invention provides an animal feed additive for immunity enhancement containing centipede grass (Eremochloa ophiuroides) extract as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물을 추가적으로 유기용매로 추출?여 제조된 유기용매 분획물을 유효성분으로 함유하는 면역 증강용 가축 사료 첨가제를 제공한다.In addition, the present invention provides an animal feed additive for immunity enhancement comprising an organic solvent fraction prepared by further extracting a centipedragus extract with an organic solvent as an active ingredient.
또한, 본 발명은 센티페드그라스 추출물의 분획물을 추가적으로 컬럼크로마토그래피로 분리한 활성분획을 유효성분으로 함유하는 면역 증강용 가축 사료 첨가제를 제공한다.In addition, the present invention provides an animal feed additive for immunity enhancement comprising an active fraction obtained by further fractionating a fraction of Sentifed glass extract by column chromatography.
본 발명의 센티페드그라스(centipede grass, Eremochloa ophiuroides) 추출물, 이의 분획물 및 상기 분획물로부터 분리한 활성분획이 면역 반응과 관련된 사이토카인(IL-6, IL-10) 및 STAT3의 인산화를 억제함으로써, 면역 증강용 가축 사료 첨가제의 유효성분으로 유용하게 사용할 수 있다.
The centipede grass of the present invention ( Eremochloa ophiuroides extract, its fractions and the active fractions isolated from the fractions inhibit the phosphorylation of cytokines (IL-6, IL-10) and STAT3 associated with the immune response and thus are useful as active ingredients in animal feed additives for immunity enhancement Can be used.
상기의 동물사료 첨가제용 조성물은 20 내지 90% 고농축액이거나 분말 또는 과립형태이다.The composition for animal feed additives is in the form of a 20 to 90% high concentration concentrate or a powder or granule.
본 발명의 동물사료 첨가제 조성물은 구연산, 후말산, 아디픽산, 젖산, 사과산 등의 유기산이나 인산나트륨, 인산칼륨, 산성피로인산염, 폴리인산염(중합인산염) 등의 인산염이나 폴리페놀, 카테킨(catechin), 알파-토코페롤, 로즈마리 추출물(rosemary extract), 비타민 C, 녹차 추출물, 감초 추출물, 키토산, 탄닌산, 피틴산 등의 천연 황산화제 중 어느 하나 또는 그 이상을 추가로 포함할 수 있다.The animal feed additive composition of the present invention can be used as a feed additive composition containing phosphate or polyphenol such as citric acid, fumaric acid, adipic acid, lactic acid, malic acid, or organic acids such as sodium phosphate, potassium phosphate, acid pyrophosphate, polyphosphate (polymerized phosphate), catechin, , Alpha-tocopherol, rosemary extract, vitamin C, green tea extract, licorice extract, chitosan, tannic acid, phytic acid, and the like.
상기 동물사료 첨가제용 조성물의 보조성분으로 아미노산, 무기염류, 비타민, 항생물질, 항균물질, 항산화, 항곰팡이 효소, 살아있는 미생물 제제 등과 같은 각종보조제가 곡물, 예를 들면 분쇄 또는 파쇄된 밀, 귀리, 보리, 옥수수 및 쌀; 식물성 단백질 사료, 예를 들면 평지, 콩 및 해바라기를 주성분으로 하는 것; 동물성 단백질 사료, 예를 들면 혈분, 육분, 골분 및 생선분; 당분 및 유제품, 예를 들면 각종 분유 및 유장 분말로 이루어지는 건조성분, 건조 첨가제를 모두 혼합한 후, 액체 성분과 가열 후에 액체가 되는 성분, 즉, 지질, 예를 들면 가열에 의해 임의로 액화시킨 동물성 지방 및 식물성 지방 등과 같은 주성분 이외에 영양보충제, 소화 및 흡수향상제, 성장 촉진제, 질병예방제 등과 같은 물질과 함께 사용될 수 있다.Various adjuvants such as amino acids, inorganic salts, vitamins, antibiotics, antimicrobial substances, antioxidants, antifungal enzymes, living microbial preparations and the like are used as auxiliary components of the composition for animal feed additive, for example, cereals such as ground or crushed wheat, Barley, corn and rice; Vegetable protein feedstuffs, for example, based on rapeseed, soybeans and sunflower; Animal protein feeds such as blood, meat, bone meal and fish meal; A sugar ingredient and a milk product, for example, a dry ingredient consisting of various powdered milk and whey powder, and a dry additive are all mixed and then mixed with a liquid ingredient and a component which becomes a liquid after heating, that is, an animal fat And vegetable fats and the like can be used together with materials such as nutritional supplements, digestion and absorption enhancers, growth promoters, disease prevention agents and the like.
상기 동물사료 첨가제용 조성물은 동물에게 단독으로, 또는 식용 담체 중에서 다른 사료 첨가제와 조합되어 투여될 수 있다.The composition for animal feed additives can be administered to animals alone or in combination with other feed additives in edible carriers.
또한, 상기 동물사료 첨가제용 조성물은 탑 드레싱으로서 또는 이들을 가축사료에 직접 혼합하거나 또는 사료와 별도로, 별도의 경구 제형으로, 주사 또는 경피로 또는 다른 성분과 조합하여 쉽게 투여할 수 있다. 통상적으로, 당업계에 잘 알려진 바와 같이 단독 일일 투여랑 또는 분할 일일 투여량을 사용할 수 있다.In addition, the composition for animal feed additives can be administered as top dressing or they can be mixed directly with the livestock feed or separately from the feed, in separate oral formulations, by injection or transdermal or in combination with other ingredients. Typically, a single daily dose or a divided daily dose can be used as is well known in the art.
상기 동물사료 첨가제용 조성물은 동물사료와 별도로 투여할 경우, 당업계에 잘 알려진 바와 같이 조성물의 투여 형태는 이들의 비-독성 약학적으로 허용 가능한 식용 담체와 조합하여 즉석 방출 또는 서방성 제형으로 제조할 수 있다. 이러한 식용 담체는 고체 또는 액체, 예를 들어 옥수수 전분, 락토오스, 수크로스, 콩 플레이크, 땅콩유, 올리브유, 참깨유 및 프로필렌 글리콜일 수 있다. 고체 담체가 사용될 경우, 추출물의 투여형은 정제, 캡슐제, 산제, 트로키제 또는 함당정제 또는 미분산성 형태의 탑 드레싱일 수 있다. 액체 담체가 사용될 경우, 연젤라틴 캡 슐제, 또는 시럽제 또는 액체 현탁액제, 에멀젼제 또는 용액제의 투여 형태일 수 있다. 또한, 투여 형태는 보조제, 예를 들어 보존제, 안정화제, 습윤제 또는 유화제, 용액 촉진제 등을 함유할 수 있다.When the composition for animal feed additives is administered separately from an animal feed, the dosage form of the composition, as is well known in the art, is combined with their non-toxic pharmaceutically acceptable edible carrier to produce an instant release or sustained release formulation can do. Such edible carriers may be solid or liquid, for example corn starch, lactose, sucrose, soy flakes, peanut oil, olive oil, sesame oil and propylene glycol. When a solid carrier is used, the dosage form of the extract may be a tablet, capsule, powder, troche or emulsion or top-dressing in finely divided form. When a liquid carrier is used, it may be in the form of a soft gelatin capsule, or a syrup or liquid suspension, emulsion or solution. In addition, the dosage form may contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution promoting agents and the like.
또한, 상기 동물사료 첨가제를 포함하는 사료는 동물의 식이 요구를 충족시키는데 통상적으로 사용되는 임의의 단백질-함유 유기 곡분일 수 있다. 이러한 단백질-함유 곡분은 통상적으로 옥수수, 콩 곡분 또는 옥수수/콩 곡분 믹스로 주로 구성되어 있다.In addition, the feed comprising the animal feed additive may be any protein-containing organic grain fraction commonly used to meet animal dietary needs. These protein-containing flours usually consist mainly of corn, soy flour or corn / soy flour mix.
상기의 동물사료 첨가제는 침지, 분무 또는 혼합하여 상기 동물사료에 첨가하여 이용될 수 있다.The animal feed additive may be added to the animal feed by immersion, spraying or mixing.
본 발명은 포유류, 가금 및 어류를 포함하는 다수의 동물 식에 적용할 수 있다. 보다 상세하게, 식이는 상업상 중요한 포유류, 예를 들어 돼지, 소, 양, 염소, 실험용 설치 동물(예, 랫트, 마우스, 햄스터 및 게르빌루스쥐), 모피 소유 동물(예, 밍크 및 여우), 및 동물원 동물(예, 원숭이 및 꼬리 없는 원숭이), 뿐만 아니라 가축(예, 고양이 및 개)에 사용할 수 있다. 통상적으로 상업상 중요한 가금에는 닭, 칠면조, 오리, 거위, 꿩 및 메추라기가 포함된다. 송어와 같은 상업적으로 사육되는 어류도 포함될 수 있다.The present invention is applicable to a large number of animal models including mammals, poultry, and fish. More specifically, the diets may be used in commercial mammals, such as pigs, cows, sheep, goats, laboratory animals (e.g., rats, mice, hamsters and gervilus rats), fur- , And zoo animals (e.g., monkeys and tailless monkeys), as well as livestock (eg, cats and dogs). Common commercially important poultry include chickens, turkeys, ducks, geese, pheasants and quails. Commercial breeding fish such as trout can also be included.
본 발명에 따른 동물사료 첨가제를 포함한 동물사료 배합 방법은, 상기 동물사료 첨가제를 동물사료에 건조 중량 기준으로 사료 1kg당 약 1g 내지 100g의 양으로 혼입한다.In the animal feed compounding method including the animal feed additive according to the present invention, the animal feed additive is incorporated into the animal feed in an amount of about 1 g to 100 g per 1 kg of the feed on a dry weight basis.
또한, 사료 혼합물은 완전히 혼합한 후, 성분들의 분쇄 정도에 따라 경점성의 조립 또는 과립 물질이 얻어진다. 이것을 메쉬로서 공급하거나, 또는 추가 가공 및 포장을 위해 원하는 분리된 형상으로 형성한다. 이 때, 저장 중에 분리되는 것을 방지하기 위해, 가축사료에 물을 첨가하고, 이어서 통상의 펠릿화, 팽창화, 또는 압출 공정을 거치는 것이 바람직하다. 과잉의 물은 건조 제거될 수 있다.
Also, after the feed mixture is thoroughly mixed, lightly viscous granulation or granulation material is obtained depending on the degree of crushing of the components. This is supplied as a mesh, or is formed into a desired discrete shape for further processing and packaging. At this time, in order to prevent separation during storage, it is preferable to add water to the livestock feed, followed by a usual pelleting, expansion, or extrusion process. Excess water can be dried off.
이하, 본 발명을 실시예 및 제조예에 의하여 상세히 설명한다.Hereinafter, the present invention will be described in detail with reference to Examples and Production Examples.
단, 하기 실시예 및 제조예는 본 발명을 구체적으로 예시하는 것이며, 본 발명의 내용이 실시예 및 제조예에 의해 한정되는 것은 아니다.
However, the following examples and preparation examples illustrate the present invention specifically, and the content of the present invention is not limited by the examples and the production examples.
<< 실시예Example 1> 1> 센티페드그라스Sentiped Grass 추출물의 제조 Preparation of extract
<1-1> <1-1> 센티페드그라스Sentiped Grass 메탄올 추출물의 제조 Preparation of methanol extract
센티페드그라스 종자를 일본의 Fukukaen Nursery and Blub co. Ltd로부터 수입하여 한국원자력연구원 정읍 방사선과학연구소의 포장에서 증식하여 센티페드그라스 잎 시료를 채취하였다. 채취한 잎 시료는 -80℃에 보관하여 사용하였다. 센티페드그라스 잎 5 ㎏을 100% 메탄올(MeOH) 20 ℓ를 첨가하여 믹서기로 파쇄한 후 상온에서 3일간 침지시켜 얻은 추출물을 50℃에서 감압 농축하여 210 g의 센티페드그라스 메탄올 추출물을 수득하였다(도 1).
The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. Leaf samples were stored at -80 ° C. 5 kg of centipedegrass leaves were added to 20 L of 100% methanol (MeOH), disintegrated with a blender and immersed at room temperature for 3 days. The extract was concentrated under reduced pressure at 50 캜 to obtain 210 g of a centipede glass methanol extract 1).
<1-2> <1-2> 센티페드그라스Sentiped Grass 에탄올 추출물의 제조 Preparation of ethanol extract
센티페드그라스 종자를 일본의 Fukukaen Nursery and Blub co. Ltd로부터 수입하여 한국원자력연구원 정읍 방사선과학연구소의 포장에서 증식하여 센티페드그라스 잎 시료를 채취하였다. 채취한 잎 시료는 -80℃에 보관하여 사용하였다. 센티페드그라스 잎 5 ㎏을 100% 에탄올(EtOH) 20 ℓ를 첨가하여 믹서기로 마쇄한 후 상온에서 3일간 침지시켜 얻은 추출물을 50℃에서 감압 농축하여 203 g의 센티페드그라스 에탄올 추출물을 수득하였다(도 1).
The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. Leaf samples were stored at -80 ° C. 5 kg of the centipede grass leaf was added with 20 L of 100% ethanol (EtOH), ground with a blender, immersed in the liquid at room temperature for 3 days, and concentrated under reduced pressure at 50 캜 to obtain 203 g of a centipede glass ethanol extract 1).
<1-3> <1-3> 센티페드그라스Sentiped Grass 물 추출물의 제조 Preparation of water extract
센티페드그라스 종자를 일본의 Fukukaen Nursery and Blub co. Ltd로부터 수입하여 한국원자력연구원 정읍 방사선과학연구소의 포장에서 증식하여 센티페드그라스 잎 시료를 채취하였다. 채취한 잎 시료는 -8℃에 보관하여 사용하였다. 센티페드그라스 잎 5 ㎏을 물 20 ℓ를 첨가하여 믹서기로 마쇄한 후 상온에서 3일간 침지시켜 얻은 추출물을 50℃에서 감압 농축하여 198 g의 센티페드그라스 물 추출물을 수득하였다(도 1).
The centipede grass seeds were collected from Fukukaen Nursery and Blub co. Ltd., And propagated in the packaging of the Korea Atomic Energy Research Institute 's Jeong - eup Radiation Research Institute and collected samples of centipede grass leaves. The collected leaf samples were stored at -8 ° C. 20 ℓ of water was added to 20 kg of centipedegrass leaves, and the mixture was ground with a mixer and immersed at room temperature for 3 days. The extract was concentrated under reduced pressure at 50 ° C to obtain 198 g of centipede water extract (FIG. 1).
<< 실시예Example 2> 2> 센티페드그라스Sentiped Grass 분획물의Fraction 제조 Produce
<2-1> n-<2-1> n- 헥산Hexane 및 에틸아세테이트 And ethyl acetate 분획물의Fraction 제조 Produce
상기 실시예 <1-1>에서 수득한 센티페드그라스 메탄올 추출물을 물에 현탁한 후 n-헥산, 에틸아세테이트를 각 용매로 사용하여 순차적으로 분획하여 n-헥산 분획물 104.77g, 에틸아세테이트 분획물 21g을 수득하였다(도 1).
The centipede glass methanol extract obtained in Example <1-1> was suspended in water, and then n-hexane and ethyl acetate were sequentially fractionated using each solvent to obtain 104.77 g of n-hexane fraction and 21 g of ethyl acetate fraction, (Fig. 1).
<2-2> 물 <2-2> Water 분획물의Fraction 제조 Produce
상기 실시예 <2-1>에서 n-헥산 및 에틸아세테이트(EtOAc)로 분획한 후 물에 의한 가용 분획물을 마지막으로 수득하였다(도 1).
After fractionation with n-hexane and ethyl acetate (EtOAc) in Example <2-1> above, the water-soluble fraction was finally obtained (Fig. 1).
<< 실시예Example 3> 3> 센티페드그라스Sentiped Grass 활성분획의 제조 Production of active fractions
<3-1> 에틸아세테이트-20% 메탄올 <3-1> Ethyl acetate-20% methanol 분획물의Fraction 제조 Produce
상기 <실시예 2>의 에틸아세테이트 분획물을 20% 메탄올을 용출용매로 사용하여 Flex-Colum TM(3 ㎝×25 ㎝)과 Toyoperarl HW-40(75 ㎛)를 사용하여 컬럼크로마토그래피를 실시하여 활성분획물을 수득하였다. 에틸아세테이트-20% 메탄올 9.41 g의 활성분획을 수득하였다(도 1).
The ethyl acetate fraction of Example 2 was dissolved in Flex-Colum TM (3 cm x 25 cm) using 20% methanol as elution solvent, Column chromatography was performed using Toyoperarl HW-40 (75 占 퐉) to obtain an active fraction. Ethyl acetate-20% methanol 9.41 g of an active fraction was obtained (Fig. 1).
<3-2> 에틸아세테이트-50% 메탄올 ≪ 3-2 > Ethyl acetate-50% methanol 분획물의Fraction 제조 Produce
상기 <실시예 2>의 에틸아세테이트 분획물을 50% 메탄올을 용출용매로 사용하여 Flex-Colum TM(3 ㎝×25 ㎝)과 Toyoperarl HW-40(75 ㎛)를 사용하여 컬럼크로마토그래피를 실시하여 활성분획물을 수득하였다. 에틸아세테이트-50% 메탄올 5.74 g의 활성분획을 수득하였다(도 1).
The ethyl acetate fraction of Example 2 was dissolved in Flex-Colum TM (3 cm x 25 cm) using 50% methanol as elution solvent, Column chromatography was performed using Toyoperarl HW-40 (75 占 퐉) to obtain an active fraction. Ethyl acetate - 50% methanol 5.74 g active fractions were obtained (Figure 1).
<3-3> 에틸아세테이트-70% 메탄올 <3-3> Ethyl acetate-70% methanol 분획물의Fraction 제조 Produce
상기 <실시예 2>의 에틸아세테이트 분획물을 70% 메탄올을 용출용매로 사용하여 Flex-Colum TM(3 ㎝×25 ㎝)과 Toyoperarl HW-40(75 ㎛)를 사용하여 컬럼크로마토그래피를 실시하여 활성분획물을 수득하였다. 에틸아세테이트-70% 메탄올 1 g의 활성분획을 수득하였다(도 1).
The ethyl acetate fraction of Example 2 was dissolved in Flex-Colum TM (3 cm x 25 cm) using 70% methanol as elution solvent, Column chromatography was performed using Toyoperarl HW-40 (75 占 퐉) to obtain an active fraction. Ethyl acetate - 70% methanol 1 g of active fraction was obtained (Fig. 1).
<< 실시예Example 4> 지다당류( 4> polysaccharides ( lipopolysaccharidelipopolysaccharide , , LPSLPS )에 의해 유도된 면역 반응에 ) Induced < / RTI > 센티페드그라스Sentiped Grass 추출물 또는 이의 Extract or its 분획물의Fraction 농도에 따른 사이토카인의 발현 분석 Analysis of cytokine expression by concentration
LPS에 의해 유도된 대식세포 Raw 264.7 세포 및 마우스에 센티페드그라스 추출물 또는 이의 분획물을 처리한 후, 사이토카인 활성 정도를 효소 면역 측정 분석법(enzyme-linked immunosorbent assay, Eliza assay, Diagnostic Automation/Cortez Diagnostic, INC.) 분석을 하였다. LPS-induced macrophage Raw 264.7 cells and mice were treated with Centipedgrass extract or fractions thereof and the degree of cytokine activity was determined by enzyme-linked immunosorbent assay (Eliza assay, Diagnostic Automation / Cortez Diagnostic, INC.) Analysis.
구체적으로, 대식세포 Raw 264.7 세포를 96 웰/플레이트에 배양하며, 배양조건은 각각의 웰에 만개의 대식세포를 넣고 10% FBS가 포함된 DMEM(Dulbecco's Modified Eagle's Medium) 배지 200㎕에서 5% CO2 농도, 37℃ 온도의 조건에서 배양하였다. 이후 LPS와 센티페드그라스 추출물 또는 이의 분획물을 24시간 동안 배양하였다. 24시간 뒤, 배양액을 이용하여 대식세포가 분비한 사이토카인 중 면역 반응과 관련된 인터루킨-6(interleukin-6; IL-6)와 인터루킨-10(IL-10)의 함량을 비교하였다. 또한, 마우스(ICR, The Jackson Laboratory)에 센티페드그라스 추출물이 혼합된 사료(표 1)를 2주간 섭취시킨 후, LPS를 복강투여하여 면역반응을 유도하고 혈액 내 면역반응과 관련된 사이토카인의 함량을 비교 분석하였다. 이때 사용된 사료는 센티페드그라스 메탄올 추출물에 일반사료를 침지시킨 후, 건조시켜 사료에 센티페드그라스 추출물이 흡수되도록 하여 사용하였다. 그런 다음, Asepromazine(Sigma, USA) 및 케타민(Sigma, USA)을 혼합하여 마취액을 만들고 상기의 마우스의 체장 근육에 주입하여 마취시킨 후 심장 채혈을 통하여 혈액을 채취하여 헤파린 튜브에 옮겼다. 10,000 rpm에서 10분 동안 원심분리 후 혈청 성분을 분리 채취하고 효소 면역 측정법으로 각각 사이토카인 분석 키트(cytokine assay kit, Enzo)를 이용하여 수행하였다. 항체가 흡착된 플레이트에 검체인 혈액을 넣고서 플레이트 쉐이커(plate shaker)에서 1 시간 내지 2 시간 반응시켰다. 4회 내지 5회 정도로 충분히 씻어 비특이적 반응 산물을 제거시킨 후 각 검체의 항체를 넣어 1 시간 동안 반응시켰다. 4회 내지 5회 정도로 충분히 세척한 후 HRP-컨쥬게이트(conjugate)를 넣어 1 시간 동안 2차 항체를 결합시켰다. 4회 내지 5 회 정도로 충분히 세척한 후 기질액을 넣어 효소와 반응시켰다. 항원이 존재하면 푸른색을 띠게 되며 더 이상 발색이 안 되게 반응 정지액을 첨가하여 노란색을 띠게 될 때 이것의 흡광도를 측정하여 450 nm에서 최종 농도를 측정하였다. Specifically, macrophage Raw 264.7 cells were cultured in 96 wells / plate. The culture conditions were as follows. Ten thousand macrophages were added to each well and 200 μl of DMEM (Dulbecco's Modified Eagle's Medium) medium containing 10% FBS was added with 5% CO 2 concentration and 37 占 폚. LPS and Centipedgrass extract or fractions thereof were then cultured for 24 hours. Twenty-four hours later, the contents of interleukin-6 (IL-6) and interleukin-10 (IL-10) related to the immune response among macrophages secreted by macrophages were compared using culture medium. In addition, a diet (Table 1) containing a mixture of centipedegrass extracts in a mouse (ICR, The Jackson Laboratory) was administered for 2 weeks, LPS was intraperitoneally administered to induce an immune response, and a cytokine content Respectively. At this time, the diets used were prepared by immersing a general diet in a centipede glass methanol extract, followed by drying so as to absorb the centipedragus extract in the diet. Then, anesthetic solution was prepared by mixing Asepromazine (Sigma, USA) and Ketamine (Sigma, USA), injected into the muscles of the mice and anesthetized. Blood was collected through cardiac blood collection and transferred to a heparin tube. After centrifugation at 10,000 rpm for 10 minutes, serum components were separated and collected by enzyme immunoassay using cytokine assay kit (Enzo). Blood as a specimen was put on a plate on which the antibody was adsorbed, and reacted for 1 hour to 2 hours on a plate shaker. After 4 to 5 times of washing, the nonspecific reaction products were removed, and each sample was reacted with the antibody for 1 hour. After washing thoroughly 4 to 5 times, the HRP-conjugate was added to bind the secondary antibody for 1 hour. After washing thoroughly 4 to 5 times, the substrate solution was added to react with the enzyme. When the antigen is present, it becomes blue. When it becomes yellow by adding the stopping solution so that it can not be further developed, its absorbance is measured and its final concentration is measured at 450 nm.
그 결과, 도 2에 나타낸 바와 같이, 대식세포는 LPS에 의해 면역반응이 일어나게 되고 과도한 면역반응에 의하여 IL-6 및 IL-10의 분비정도가 확연하게 올라가는 것을 확인하였다. 하지만, 센티페드그라스 추출물 또는 이의 분획물을 함께 처리한 군에서는 샘플의 처리농도에 의존적으로 면역반응에 관련된 사이토카인들의 분비정도가 현저히 저해되는 것을 확인하였다. 그 중에서도 센티페드그라스 70% 메탄올 분획물은 아주 낮은 농도에서도 그 활성을 나타내었고 100 ㎍/㎖의 농도로 처리한 군에서는 LPS를 처리하지 않은 군과 비슷한 정도의 결과 즉, 100% 면역반응을 억제시키는 것을 확인하였다. 따라서, 센티페드그라스 추출물 또는 이의 분획물은 대식세포가 LPS에 의해서 과도한 면역반응을 유의적으로 억제하는 것을 확인하였다(도 2). As a result, as shown in FIG. 2, macrophages were found to undergo an immune response by LPS, and the degree of secretion of IL-6 and IL-10 was markedly elevated by an excessive immune response. However, it was confirmed that the degree of secretion of the cytokines involved in the immune response was significantly inhibited in the group treated with the centipede grass extract or the fraction thereof, depending on the treatment concentration of the sample. Among them, the 70% methanol fraction of Sentifedgrass showed its activity even at a very low concentration. In the group treated with 100 ㎍ / ㎖, the result similar to that of the group not treated with LPS, that is, suppressed the 100% immune response Respectively. Thus, the centipedegrass extract or its fractions confirmed that macrophages significantly inhibited excessive immune responses by LPS (Fig. 2).
또한, 도 3에 나타낸 바와 같이, ICR 마우스에 LPS를 복강투여하게 되면 면역반응이 일어나서 이에 관련된 사이토카인의 분비가 일어난다. 이 중 IL-6 및 IL-10의 혈중 함량이 크게 증가하였다. 하지만, 2주간 센티페드그라스 추출물이 함유된 사료를 먹인 ICR 마우스는 LPS의 복강투여로 인한 IL-6 및 IL-10의 증가를 50% 수준으로 억제하는 것을 확인하였다. 또한, LPS를 복강투여하지 않고 센티페드그라스 추출물이 함유된 사료만 먹인 마우스에서 정상군과 비슷한 결과를 나타내므로써 센티페드그라스 추출물은 LPS 복강투여에 의한 동물의 체내 면역반응을 억제하여 마우스에게 다른 영향을 미치지 않는 것을 확인하였다(도 3).
In addition, as shown in FIG. 3, administration of LPS intraperitoneally to an ICR mouse results in an immune response and secretion of cytokines associated therewith. Serum levels of IL-6 and IL-10 were significantly increased. However, ICR mice fed diets containing two weeks of Centipedgrass extract inhibited the increase in IL-6 and IL-10 levels induced by intraperitoneal administration of LPS to 50%. In addition, LPS was not administered intraperitoneally, and mice fed only the diet containing Sentifedgrass extract showed similar results to those of the normal group. Thus, the centipedegrass extract inhibited the animal's immune response by intraperitoneal administration of LPS, (Fig. 3).
센티페드그라스 추출물 사료 섭취량Average daily ICR mice
Sentipedgrass Extract Feed Intake
<< 실시예Example 5> 5> LPSLPS 에 의해 유도된 면역 반응에 In the immune response induced by 센티페드그라스Sentiped Grass 추출물 또는 이의 Extract or its 분획물의Fraction 농도에 따른 단백질의 발현 분석 Analysis of Protein Expression by Concentration
LPS에 의해 유도된 면역반응에 센티페드그라스 추출물 또는 이의 분획물을 처리한 후, 면역반응과 관련된 단백질의 발현을 확인하기 위하여 웨스턴 블롯(western blot)을 실시하였다.The LPS-induced immune response was treated with Centipedgrass extract or its fraction and then subjected to western blotting to confirm the expression of proteins involved in the immune response.
구체적으로, 대식세포를 2 ×106개 세포가 되도록 60 mm 디쉬에 3 ㎖씩 분주하고, 37℃, 5% CO₂인큐베이터에서 6시간 배양하여 세포를 디쉬에 부착시킨 후, LPS와 상기 <실시예 3>에서 수득한 센티페드그라스 70% 메탄올을 농도에 따라 처리한 다음 각 세포를 모아 프로테나아제 억제제(1 mM phenylmethylsulfonyl fluoride, 0.1 okadaic acid, 각각 10 /㎖ 의 aprotinin, leupeptin 및 pepstatin)를 포함하는 용해 완충액(150 mM NaCl, 10 mM Tris-HCl, 1 mM EGTA, 1 mM EDTA, 0.2 mM sodium vanadate, 0.5% Nonidet P-40)에 부유시킨 다음 얼음에 30분간 방치한 후 초음파 분해하였다. 상기 각 세포 용해물을 2 × SDS 정지 완충액(2% SDS, 5 mM EGTA, 5 mM EDTA, 25 mM dithiothreitol, 10% glycerol, 0.01% bromphenol blue, and 0.25 M Tris-Cl, pH 6.8)에 희석시키고 5분 동안 100℃에서 끓였다. 각 세포 용해물에서 동량의 단백질을 10% SDS-PAGE로 분리하고, 니트로셀룰로오스 막으로 이동시켰다. 니트로셀룰로오스 막의 비특이적 항체 결합을 막기 위하여 1시간 동안 블로킹시킨 다음, PBS 용액으로 2회 세척하였다. 상기 막에 1차 항체들(p-STAT3, STAT3, pErk, Erk, p-Akt, Akt, COX-2 및 GAPDH)을 1:100-1:1000의 비율로 희석하고 상온에서 2시간동안 일정한 방향으로 흔들어 주었다. 다시 2-3회 PBS로 세척한 다음 탐식자가 결합된 2차 항체를 같은 비율로 희석하여 상온에서 2시간 동안 반응시킨 후, X-ray 필름에 현상하여 각 단백질의 발현량을 확인하였다.Specifically, 3 ml of each macrophage was dispensed into a 60 mm dish at a density of 2 × 10 6 cells and incubated in a 5
그 결과, STAT3는 IL-6 발현에 깊이 관여하는 단백질 중 하나로 센티페드그라스 70% 메탄올 분획물을 20 ㎍/㎖ 농도 이상으로 처리한 군에서 STAT3의 인산화가 현저하게 감소하였다. 하지만, Erk, Akt와 같은 단백질의 발현이나 인산화에는 영향을 미치지 않았다. COX-2 단백질의 경우, 센티페드그라스 70% 메탄올 분획물을 높은 농도인 100 ㎍/㎖으로 처리한 군에서만 발현이 감소하는 것을 확인하였다(도 4). As a result, STAT3 was one of the proteins deeply involved in the expression of IL-6, and STAT3 phosphorylation was markedly decreased in the group treated with 70% methanol fraction of centipedegrass at a concentration of 20 μg / ml or more. However, it did not affect the expression or phosphorylation of proteins such as Erk and Akt. In the case of the COX-2 protein, the expression was decreased only in the group treated with 70% methanol fraction of Centipedgrass at a high concentration of 100 占 퐂 / ml (FIG. 4).
따라서, 센티페드그라스 70% 메탄올 분획물은 STAT3가 인산화되는 것을 저해하여 면역반응에 중요한 사이토카인 중 하나인 IL-6의 발현이 감소시킴으로써 면역 증강에 효과가 있음을 확인하였다.
Therefore, the 70% methanol fraction of Sentifedgrass inhibited the phosphorylation of STAT3, indicating that IL-6, which is one of the important cytokines in the immune response, is reduced and thus the immune enhancement is effective.
<< 제조예Manufacturing example 1> 1> 센티페드그라스Sentiped Grass 추출물을 유효성분으로 함유하는 면역증강용 For enhancing immunity containing extract as active ingredient 약제의 제조Manufacture of medicines
<< 제제예Formulation example 1> 약학적 제제의 제조 1> Preparation of pharmaceutical preparations
<1-1><1-1> 산제의Sanje 제조 Produce
실시예 <1-1> 센티페드그라스 메탄올 추출물 0.1 gExample < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract
유당 1.5 gLactose 1.5 g
탈크 0.5 gTalc 0.5 g
상기의 성분들을 혼합하고 기밀포에 충진하여 산제를 제조하였다.
The above ingredients were mixed and filled in an airtight container to prepare powders.
<1-2> 정제의 제조<1-2> Preparation of tablets
본 발명의 센티페드그라스 추출물을 유효성분으로 함유하는 정제는 하기 표 2의 조성과 같이 제조하였다.
Tablets containing the centipedragus extract of the present invention as an active ingredient were prepared according to the composition shown in Table 2 below.
본 발명의 추출물 250 중량부, 락토오스 175.9 중량부, 감자전분 180 중량부 및 콜로이드성 규산 32 중량부와 혼합하였다. 상기 혼합물에 10% 젤라틴 용액을 첨가시킨 후, 분쇄하여 14 매쉬체를 통과시켰다. 이것을 건조하고 여기에 감자전분 160 중량부, 활석 50 중량부 및 스테아린산 마그네슘 5 중량부를 첨가하여 얻은 혼합물을 정제로 제조하였다.
250 parts by weight of the extract of the present invention, 175.9 parts by weight of lactose, 180 parts by weight of potato starch and 32 parts by weight of colloidal silicic acid. 10% gelatin solution was added to the mixture, followed by pulverization and passed through a 14-mesh sieve. This was dried, to which 160 parts by weight of potato powder, 50 parts by weight of talc and 5 parts by weight of magnesium stearate were added, and the resulting mixture was purified.
<1-3><1-3> 캡슐제의 제조 Preparation of capsules
실시예 <1-1> 센티페드그라스 메탄올 추출물 0.1 gExample < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract
옥수수전분 5 gCorn starch 5 g
카르복시 셀룰로오스 4.9 g4.9 g of carboxycellulose
상기의 성분들을 혼합하여 분말을 제조한 후, 상기 분말을 통상의 캡슐제의 제조방법에 따라 경질 캡슐에 충전하여 캡슐제를 제조하였다.
After mixing the above components to prepare a powder, the powder was filled in a hard capsule according to a conventional preparation method of a capsule to prepare a capsule.
<1-4> 주사제의 제조≪ 1-4 > Preparation of injection
실시예 <1-1> 센티페드그라스 메탄올 추출물 0.1 gExample < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract
주사용 멸균 증류수 적량Sterile sterilized water for injection
pH 조절제 적량pH adjuster
통상의 주사제의 제조방법에 따라 1 앰플 당(2 ㎖) 상기의 성분 함량으로 제조하였다.
(2 ml) per ampoule according to the usual injection preparation method.
<1-5><1-5> 액제의Liquid 제조 Produce
실시예 <1-1> 센티페드그라스 메탄올 추출물 0.1 gExample < 1-1 > A mixture of 0.1 g of Centipede Grass methanol extract
이성화당 10 g10 g per isomer
만니톨 5 g5 g mannitol
정제수 적량Purified water quantity
통상의 액제의 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고, 레몬향을 적량 가한 다음 상기의 성분을 혼합하였다. 그 다음 정제수를 가하여 전체 100 로 조절한 후 갈색병에 충진하여 멸균시켜 액제를 제조하였다.
Each component was dissolved in purified water in accordance with a conventional method for producing a liquid agent, and the lemon flavor was added in an appropriate amount, followed by mixing the above components. Then, purified water was added to adjust the total amount to 100, and the solution was filled in a brown bottle and sterilized to prepare a liquid preparation.
<1-6> 시럽제의 제조≪ 1-6 > Preparation of syrup preparation
본 발명의 센티페드그라스 추출물을 유효성분으로 함유하는 시럽제는 하기 표 3의 조성과 같이 제조하였다.
Syrup preparations containing the centipedragus extract of the present invention as an active ingredient were prepared according to the composition shown in Table 3 below.
<< 제조예Manufacturing example 2> 2> 센티페드그라스Sentiped Grass 추출물을 유효성분으로 함유하는 면역증강용 건강식품의 제조 Preparation of a health food for immunity enhancement containing an extract as an active ingredient
<2-1> 밀가루 식품의 제조<2-1> Production of flour food
상기 실시예 <1-3>의 센티페드그라스 물 추출물 0.5 ~ 5.0 중량부를 밀가루에 첨가하고, 이 혼합물을 이용하여 빵, 케이크, 쿠키, 크래커 및 면류를 제조하였다.
0.5-5.0 parts by weight of the centipedglass water extract of Example <1-3> was added to wheat flour, and the mixture was used to prepare bread, cake, cookies, crackers and noodles.
<2-2> <2-2> 스프soup 및 육즙( And juicy ( graviesgravies )의 제조)
상기 실시예 <1-3>의 센티페드그라스 물 추출물 0.1 ~ 5.0 중량부를 스프 및 육즙에 첨가하여 건강 증진용 육가공 제품, 면류의 수프 및 육즙을 제조하였다.
0.1 to 5.0 parts by weight of the centipedglass water extract of Example <1-3> was added to the soup and the juice to prepare a health promotion meat product, noodle soup and juice.
<2-3> 그라운드 <2-3> Ground 비프(ground beef)의Beef 제조 Produce
상기 실시예 <1-3>의 센티페드그라스 물 추출물 10 중량부를 그라운드 비프에 첨가하여 건강 증진용 그라운드 비프를 제조하였다.
10 parts by weight of the centipede water extract of Example <1-3> was added to the ground beef to prepare ground beef for health promotion.
<2-4> 유제품(<2-4> Dairy products ( dairydairy productsproducts )의 제조)
상기 실시예 <1-3>의 센티페드그라스 물 추출물 5 ~ 10 중량부를 우유에 첨가하고, 상기 우유를 이용하여 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.
5-10 parts by weight of the centipedglass water extract of Example <1-3> was added to milk, and various dairy products such as butter and ice cream were prepared using the milk.
<2-5> <2-5> 선식의Solar 제조 Produce
현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화시켜 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.
검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.
상기 실시예 <1-3>의 센티페드그라스 물 추출물을 진공 농축기에서 감압농축하고, 분무, 열풍건조기로 건조하여 얻은 건조물을 분쇄기로 입도 60 메쉬로 분쇄하여 건조분말을 얻었다.The centipede water extract of Example <1-3> was concentrated under reduced pressure in a vacuum concentrator, dried by spraying and dried in a hot air drier, and pulverized to a size of 60 mesh with a pulverizer to obtain a dry powder.
상기에서 제조한 곡물류, 종실류 및 실시예 <1-3>의 센티페드그라스 물 추출물을 다음의 비율로 배합하여 제조하였다.The above-prepared cereals, seeds and centipede water extract of Example <1-3> were prepared in the following proportions.
곡물류(현미 30 중량부, 율무 15 중량부, 보리 20 중량부),(30 parts by weight of brown rice, 15 parts by weight of yulmu, 20 parts by weight of barley)
종실류(들깨 7 중량부, 검정콩 8 중량부, 검정깨 7 중량부),Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)
실시예 <1-3>의 추출물(3 중량부),The extract (3 parts by weight) of Example <1-3>,
영지(0.5 중량부),(0.5 part by weight),
지황(0.5 중량부)
(0.5 parts by weight)
<< 제조예Manufacturing example 3> 3> 센티페드그라스Sentiped Grass 추출물을 유효성분으로 함유하는 면역증강용 기능성 음료의 제조 Preparation of a functional beverage for immunity enhancement containing an extract as an active ingredient
<3-1> 건강 음료의 제조<3-1> Production of health drinks
액상과당(0.5%), 올리고당(2%), 설탕(2%), 식염(0.5%), 물(75%)과 같은 부재료와 본 발명의 센티페드그라스 추출물 5 g을 균질하게 배합하여 순간 살균을 한 후 이를 유리병, 패트병 등 소포장 용기에 포장하여 제조하였다.
(5%) of the centipede glass extract of the present invention was homogeneously mixed with a mixture of liquid fructose (0.5%), oligosaccharide (2%), sugar (2%), salt (0.5% And packaged in small containers such as glass bottles and plastic bottles.
<3-2> 야채 주스의 제조<3-2> Preparation of vegetable juice
본 발명의 센티페드그라스 추출물 5 g을 토마토 또는 당근 주스 1,000 ㎖에 가하여 야채 주스를 제조하였다.
5 g of the centipede glass extract of the present invention was added to 1,000 ml of tomato or carrot juice to prepare vegetable juice.
<3-3> 과일 주스의 제조<3-3> Production of fruit juice
본 발명의 센티페드그라스 추출물 1 g을 사과 또는 포도 주스 1,000 ㎖ 에 가하여 과일 주스를 제조하였다.
1 g of the centipede glass extract of the present invention was added to 1,000 ml of apple or grape juice to prepare fruit juice.
<< 제조예Manufacturing example 4> 4> 센티페드그라스Sentiped Grass 추출물을 유효성분으로 함유하는 면역증강용 사료 및 사료 첨가제의 제조 Preparation of immunostimulating feeds and feed additives containing an extract as an active ingredient
<4-1> 가축 사료의 제조<4-1> Production of livestock feed
본 발명의 센티페드그라스 추출물을 유효성분으로 함유하는 가축 사료는 하기 표 4의 조성과 같이 제조하였다.
The livestock feed containing the centipede grass extract of the present invention as an active ingredient was prepared according to the composition shown in Table 4 below.
<4-2> 가축 사료 첨가제의 제조<4-2> Preparation of animal feed additive
본 발명의 센티페드그라스 추출물 0.5 g을 액상 상태의 비타민, 미네랄 혼합물, 염화콜린, 식물성 지방, 트레오닌을 추가하여 고르게 혼합해 제조하였다.
0.5 g of the centipede glass extract of the present invention was prepared by mixing liquid vitamins, minerals, choline chloride, vegetable fats and threonine evenly.
Claims (10)
An immunomodulating health food containing the extract of Eremochloa ophiuroides as an active ingredient.
The method of claim 1, wherein the grass extract is fed centimeters of water, C 1 to immunity enhancement for healthy foods, characterized in that extraction with a lower alcohol or a mixed solvent of a C 2.
The health food according to claim 2, wherein the lower alcohol is ethanol or methanol.
The centipedegrass extract further comprises an organic solvent fraction prepared by extraction with an organic solvent as an active ingredient.
The health food according to claim 4, wherein the organic solvent is n-hexane or ethyl acetate.
5. The composition according to claim 4, wherein the fraction is an ethyl acetate fraction obtained by extracting Centipedgrass extract with n-hexane and then extracting the water layer with ethyl acetate.
A health food for immune enhancement comprising an active fraction which is obtained by further adsorbing an ethyl acetate fraction of a centipedragus extract by column chromatography and using methanol as an eluting solvent as an active ingredient.
A feed additive for immune enhancement containing a centipede grass extract as an active ingredient.
The centipede-grass extract further comprises an organic solvent fraction prepared by extraction with an organic solvent as an active ingredient.
An animal feed additive for enhancing immunity comprising an active fraction which is obtained by further adsorbing an ethyl acetate fraction of a centipedlass extract to column chromatography and using methanol as an eluting solvent.
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KR20110040375A (en) * | 2009-10-14 | 2011-04-20 | 한국원자력연구원 | Method for removing the pigment of centipede grass extract using irradiation |
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WO2018110875A1 (en) | 2016-12-12 | 2018-06-21 | Korea Atomic Energy Research Institute | Pharmaceutical composition for the prevention or treatment of alopecia comprising eremochloa ophiuroides extract or fractions thereof as an active ingredient |
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CN113933425A (en) * | 2021-10-12 | 2022-01-14 | 通标标准技术服务(上海)有限公司 | Method for measuring content of acetophenone azine in plastic, chloroprene rubber and foam |
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