KR100445993B1 - Food additive containing cacao bean or husk fraction extract - Google Patents
Food additive containing cacao bean or husk fraction extract Download PDFInfo
- Publication number
- KR100445993B1 KR100445993B1 KR10-2004-0047864A KR20040047864A KR100445993B1 KR 100445993 B1 KR100445993 B1 KR 100445993B1 KR 20040047864 A KR20040047864 A KR 20040047864A KR 100445993 B1 KR100445993 B1 KR 100445993B1
- Authority
- KR
- South Korea
- Prior art keywords
- fractions
- cacao
- inflammation
- weight
- cacao beans
- Prior art date
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Classifications
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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
-
- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/30—Removing undesirable substances, e.g. bitter substances
- A23L11/32—Removing undesirable substances, e.g. bitter substances by extraction with solvents
-
- 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
- A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
- A23L11/30—Removing undesirable substances, e.g. bitter substances
- A23L11/34—Removing undesirable substances, e.g. bitter substances using chemical treatment, adsorption or absorption
-
- 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
-
- 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
- A23V2300/00—Processes
- A23V2300/14—Extraction
-
- 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
- A23V2300/00—Processes
- A23V2300/18—Fractionation
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- Life Sciences & Earth Sciences (AREA)
- Botany (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Agronomy & Crop Science (AREA)
- Mycology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제에 관한 것으로서, 더욱 상세하게는 세균이 분비하는 내독소에 의해 발생하는 소화기계 염증질환 및 여러 피부염증 유발 물질들에 의해 발생되는 원발성 접촉피부염에 대해 뛰어난 억제 효능을 확인함으로써, 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제에 관한 것이다.The present invention relates to an anti-inflammatory food additive containing cacao beans or bean skin fractions, and more particularly, is caused by digestive system inflammatory diseases and various dermatitis-causing substances caused by endotoxins secreted by bacteria. By identifying excellent inhibitory efficacy against primary contact dermatitis, it relates to an anti-inflammatory food additive containing cacao beans or soybean skin fractions.
Description
본 발명은 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제에 관한 것으로서, 더욱 상세하게는 세균이 분비하는 내독소에 의해 발생하는 소화기계 염증질환 및 여러 피부염증 유발 물질들에 의해 발생되는 원발성 접촉피부염에 대해 뛰어난 억제 효능을 확인함으로써, 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제에 관한 것이다.The present invention relates to an anti-inflammatory food additive containing cacao beans or bean skin fractions, and more particularly, is caused by digestive system inflammatory diseases and various dermatitis-causing substances caused by endotoxins secreted by bacteria. By identifying excellent inhibitory efficacy against primary contact dermatitis, it relates to an anti-inflammatory food additive containing cacao beans or soybean skin fractions.
초콜릿 및 코코아의 원료인 카카오는 학명이 데오브로마 카카오 엘.(Theobroma cacao L)로, 신의 음식이라는 뜻을 내포하고 있다. 이러한 카카오의 원산지는 중앙아메리카에서 남아메리카의 북부에 걸친 지역으로 추정되며, 중앙아메리카의 여러 나라에서는 기원전 오래 전부터 재배되어 옥수수와 함께 매우 귀중한 음식으로 취급되어 왔으며, 현재는 남녀노소, 동서양을 막론하고 전세계적인 대표적인 기호식품으로 자리를 잡고 있다. 초콜릿의 원료로 제공되는 카카오 콩은 산지에 따라 차이가 있으나, 공통적으로 식이섬유가 풍부할 뿐만 아니라, 폴리페놀 등이 상당량 함유되어 있다.Cacao, the raw material of chocolate and cocoa, has the scientific name Theobroma cacao L, meaning divine food. The origin of cacao is estimated to be from Central America to the northern part of South America, and has been cultivated for a long time in BC and has been treated with corn as a very valuable food. It is settled as a typical sign food. Cacao beans, which are used as raw materials for chocolate, vary depending on the country of origin, but are not only rich in dietary fiber, but also contain significant amounts of polyphenols.
한편, 카카오 콩은 초콜릿의 생산공정 중 탈각되어 알맹이(nib)는 초콜릿 또는 코코아의 원료로 사용되고, 부산물로 나오는 껍질(전체 카카오 콩에 대하여 약 15 중량%)은 거의 대부분 폐기되고 있는 실정이다. 다시 말하면, 전세계 카카오 콩의 생산량은 약 250만톤으로, 약 40만톤에 이르는 엄청난 양의 카카오 콩껍질이 폐기되고 있으며, 1990년 관세청 무역통계 연보에 따르면, 우리 나라에서도 약 600톤의 카카오 콩껍질이 폐기물로 배출되고 있는 것으로 알려지고 있다. 그러나, 이러한 카카오 콩껍질에는 카카오 콩과 마찬가지로 식이섬유가 풍부할 뿐만 아니라, 폴리페놀 등이 상당량 함유되어 있다.On the other hand, cacao beans are shelled out during the production of chocolate so that nibs are used as raw materials for chocolate or cocoa, and shells (about 15% by weight of the total cacao beans) that are produced as by-products are almost discarded. In other words, the world produces about 2.5 million tons of cacao beans, about 400,000 tons of cacao beans are being discarded, and according to the 1990 Customs Statistics Korea, about 600 tons of cacao beans It is known that it is being discharged as waste. However, like cacao beans, such cacao beans are not only rich in dietary fiber, but also contain a significant amount of polyphenols.
따라서, 카카오 콩에 있는 폴리페놀(polyphenol)의 기능성을 이용하는 방법이 강구되어 왔다. 카카오 콩의 폴리페놀 및 분획물이 백혈병, 전립선암, 결장암, 신장암, 폐암, 유방암 세포를 사멸시키며, 또한 토포아이소머라제 Ⅱ를 저해시키는 것을 밝힘으로써 다양한 구조의 폴리페놀들이 항암활성 및 여러 생리활성을 가질 것으로 생각되어 이들 폴리페놀의 구조 및 용도에 대한 특허를 청구한 바 있다[대한민국 특허 출원번호 제 1998-707952호].Thus, methods have been devised to exploit the functionality of polyphenols in cacao beans. Polyphenols of various structures have been shown to inhibit the activity of leukemia, prostate cancer, colon cancer, kidney cancer, lung cancer, breast cancer and also inhibit topoisomerase II. It has been claimed to have a patent for the structure and use of these polyphenols (Korean Patent Application No. 1998-707952).
그러나, 본 발명자들은 상기에 특허 청구한 카카오 콩의 폴리페놀들의 구조 및 이에 따른 생리활성에 대한 특허가 출원되기 훨씬 이전에, 콩껍질 폴리페놀 추출물이 초콜릿 생산 중 부산물로 얻어지는 것을 사용하므로 가격이 싸다는 것과,폴리페놀도 프로시아니딘 A(procyanidin A), 프로시아니딘 C-1(procyanidin C-1) 등 중합체(polymer)를 주성분으로 하므로 효소저해 활성도 매우 높아 충치 원인 효소인 글루코실트랜스퍼라제의 활성을 저해한다는 사실을 발견하고, 카카오 콩껍질 추출물을 제조하고, 이를 배합한 츄잉껌을 제조한 바 있다[대한민국 특허 제 45179 호, 1991; 미국 특허 제 4908212호, 1990]. 최근에는 폴리페놀의 함량을 증가시켜 글루코실트랜스퍼라제의 저해 활성을 더욱 증강시킨 카카오 콩껍질 분획물을 제조하는 방법을 개발하여 특허 등록된 바 있다[미국 특허 제 6159451호, 2000].However, the inventors of the present invention are inexpensive since they use soybean peel polyphenol extracts obtained as by-products during chocolate production, long before the patents on the structure and the physiological activity of the polyphenols of cacao beans, which are claimed above, are filed. In addition, polyphenols are mainly composed of polymers such as procyanidin A and procyanidin C-1, and thus have high enzymatic inhibitory activity, which inhibits the activity of glucosyltransferase, a decay-causing enzyme. In fact, the cacao bean husk extract was prepared, and chewing gum containing the same was prepared [Korean Patent No. 45179, 1991; U.S. Patent 4908212, 1990. Recently, patents have been developed by increasing the content of polyphenols to prepare cacao bean husk fractions which further enhance the inhibitory activity of glucosyltransferases (US Pat. No. 6159451, 2000).
상기에서 서술한 바와 같이 카카오 콩 및 콩껍질에는 식이섬유, 다양한 구조의 폴리페놀 및 페놀릭 산(phenolic acid), 데오브로민 및 카페인과 같은 메틸잔틴(methylxanthines)등 여러 생리활성을 가지는 성분이 혼합되어 있으므로, 실제로 폴리페놀의 특정 구조에 의해 생리활성을 나타내는 것이 아니며, 또한 이들의 특정 구조만을 분리하여 상업적으로 이용하는 것은 제조 비용이 너무 비싸서 이용이 불가능할 것이다. 따라서, 이미 기존에 특허 등록[미국 특허 제 6159451호, 2000]된 바와 같이 간단한 분리 공정에 의해 여러 생리활성을 크게 증강시킨 분획물을 이용하는 것이 현실적일 것이다.As mentioned above, cacao beans and bean husks contain various physiological activities such as dietary fiber, polyphenols of various structures, and phenolic acid, methylxanthines such as deobromine and caffeine. As a result, the specific structures of the polyphenols do not actually exhibit physiological activity, and the separation and commercial use of only these specific structures may be too expensive to use. Therefore, it would be practical to use fractions that greatly enhanced several physiological activities by a simple separation process as already registered in the patent (US Pat. No. 6,159,451, 2000).
염증이란 세균 유래의 농양으로서 조직에 침입하여 증식된 화농균류를 백혈구가 왕성히 탐식하여, 그 결과 생기는 백혈구의 사해, 균에 의해 장해를 받은 조직, 세포파괴물 등이 조직내 융해하여 농이 축적된 곳의 병리적 상태를 말하며, 소염작용이라 함은 침입균의 증식을 항균제 등에 의하여 억제한다든지, 혹은 농양중에 축적된 이물을 탐식하는 마크로파지(macrophage)를 활성화하여 그것들을 소화,배설하는 기능을 높여서 농양의 치료과정이 촉진되는 것을 말한다. 일반적으로 염증 반응은 생체의 세포나 조직에 어떠한 기질적 변화를 가져오는 침습이 가해질 때 그 손상부위를 수복 재생하려고 하는 생체의 방어 반응과정이다. 따라서, 이러한 일련의 반응에는 국소의 혈관, 체액의 각종 조직세포, 면역관여 세포 등이 포함된다고 한다.Inflammation is a bacterium-derived abscess that actively infiltrate tissues with purulent fungi that white blood cells devour, resulting in the death of white blood cells, tissues damaged by bacteria, and cell debris, which accumulate in the tissues. It refers to the pathological condition of the place, and the anti-inflammatory effect is to inhibit the growth of invading bacteria by antibacterial agents, or to activate the macrophage that phages foreign matter accumulated in the abscess, thereby enhancing the ability to digest and excrete them. Abscess is the process of treatment. In general, an inflammatory response is a defense process of a living body that attempts to repair and repair the damaged area when an invasion that causes some organic change in a cell or tissue of the living body is applied. Therefore, such a series of reactions include local blood vessels, various tissue cells of body fluids, immune-involved cells, and the like.
현재까지 항염증 작용물질들은 급성염증에서 손상된 조직의 세포나 염증에 관여하는 세포 또는 주화인자에 의해 유주된 백혈구의 세포막으로부터 물리화학적 및 생리적 자극에 의해 활성화된 프로스타글란딘류의 생합성을 억제하는 약물들이었다(Moyazawa, K and Mikami, T., Japan J. Pharmacol., 38, 199, 1985). 그러나 이런 약물들은 급성염증에서의 증상들에 대해서는 치료효과를 가지고 있지만 류마티스 관절염과 같은 만성염증에서는 1차적인 염증현상만을 완화할 뿐이며, 면역학적 치료효과는 기대할 수 없다고 보고되고 있다. 한편 근래에는 아라키돈 산 (arachidonic acid)이 리폭시게네이즈(lipoxygenase)에 의해 생성되는 물질인 히드록시에이코사테트라에노익 산(HETE; hydroxyeicosatetraenoic acid), 류코트리엔 B4, 류코트리엔 C4 등에 관하여 연구가 활발하게 진행되고 있으며, 이는 만성염증에서 어느 정도 효과를 보이고 있다고 한다(Huskisson, E.D. et al., Inflamma- tion, Mechanism and Treatment, MTP, Lancaster, England, p.55, 1980).To date, anti-inflammatory agents have been shown to inhibit the biosynthesis of prostaglandins activated by physicochemical and physiological stimuli from the cell membranes of damaged tissues or leukocytes inoculated by chemotactic factors in acute inflammation. (Moyazawa, K and Mikami, T., Japan J. Pharmacol., 38, 199, 1985). However, these drugs have a therapeutic effect on the symptoms of acute inflammation, but in chronic inflammation such as rheumatoid arthritis, only the primary inflammation is reported, and immunological therapeutic effects are not expected. Recently, researches on arachidonic acid have been actively conducted on hydroxyeicosatetraenoic acid (HETE), leukotriene B4, and leukotriene C4, which are substances produced by lipoxygenase. This has been shown to have some effect on chronic inflammation (Huskisson, ED et al., Inflammation, Mechanism and Treatment, MTP, Lancaster, England, p.55, 1980).
따라서 천연식물 추출물을 포함하여 별도의 정제 과정 없이 안전하게 섭취할 수 있으며, 염증을 억제할 수 있고, 용이하게 식품에 이용할 수 있는 물질에 대한 연구가 필요한 실정이다.Therefore, it is necessary to study the substances that can be safely ingested without additional purification process, including natural plant extracts, can inhibit inflammation, and can be easily used in food.
그러나, 현재까지 카카오 콩 및 카카오 콩껍질 추출물의 세균의 내독소 및 염증 유발 물질들에 의해 발생되는 염증에 대한 억제 효과에 관한 보고는 전무하다.However, there are no reports on the inhibitory effect of cacao beans and cacao bean husk extracts on inflammation caused by bacterial endotoxins and inflammation-causing substances.
이에, 본 발명자들은 카카오 콩 또는 콩껍질 분획물의 생리활성을 다각도로 연구하던 중, 이들 분획물들이 동물 실험에서 세균의 내독소에 의해 발생하는 소화기계 염증질환 및 원발성 접촉피부염에 대해 뛰어난 억제 효능을 밝힘으로써, 본 발명을 완성하게 되었다.Therefore, the inventors of the present invention, while studying the physiological activity of the cacao beans or soybean shell fractions from various angles, and revealed that these fractions have excellent inhibitory effects against digestive system inflammatory diseases and primary contact dermatitis caused by bacterial endotoxin in animal experiments. Thus, the present invention has been completed.
따라서, 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제를 제공하는데 그 목적이 있다.Accordingly, an object of the present invention is to provide an anti-inflammatory food additive containing cacao beans or a legume fraction.
도 1은 카카오 콩 분획물을 쥐에 경구 투여했을 때, 세균의 내독소인 LPS(lipopolysaccharide)에 의한 쥐 치사율을 나타낸 그래프이다.1 is a graph showing the mouse mortality rate by the endotoxin of bacteria, LPS (lipopolysaccharide) when orally administered cacao beans fractions to mice.
도 2는 카카오 콩껍질 분획물을 쥐에 경구 투여했을 때, 세균의 내독소인 LPS에 의한 쥐 치사율을 나타낸 그래프이다.2 is a graph showing the mouse mortality rate by LPS, which is a bacterial endotoxin when orally administered cacao bean husk fraction to mice.
도 3은 카카오 콩 분획물을 쥐에 경구 처치했을 때, 피부염증 유발 물질인 TPA(12-O-tetradecanoylphorbol-13-acetate)에 의한 쥐의 귀 염증 억제율을 나타낸 그래프이다.Figure 3 is a graph showing the inhibition rate of ear inflammation in rats by oral treatment of cacao bean fraction in rats, TPA (12- O- tetradecanoylphorbol-13-acetate) which is a dermatitis-inducing substance.
도 4는 카카오 콩껍질 분획물을 쥐에 경구 처치했을 때, 피부염증 유발 물질인 TPA에 의한 쥐의 귀 염증 억제율을 나타낸 그래프이다.4 is a graph showing the inhibition rate of ear inflammation in rats by TPA, a dermatitis-inducing substance, when cacao bean skin fractions were orally treated in rats.
도 5는 카카오 콩 분획물을 쥐에 경구 처치했을 때, 피부염증 유발 물질인 크로톤오일(croton oil)에 의한 쥐의 귀 염증 억제율을 나타낸 그래프이다.FIG. 5 is a graph showing the ear inflammation inhibition rate of rats by croton oil, a skin inflammation-causing substance, when cacao bean fractions were orally treated in mice.
도 6은 카카오 콩껍질 분획물을 쥐에 경구 처치했을 때, 염증 유발 물질인 크로톤오일에 의한 쥐의 귀 염증 억제율을 나타낸 그래프이다.FIG. 6 is a graph showing the inhibition rate of ear inflammation in rats by croton oil, an inflammation-causing substance, when cacao bean skin fractions were orally treated in mice.
도 7은 카카오 콩 분획물을 쥐에 경구 처치했을 때, 염증 유발 물질인 디스라놀(dithranol)에 의한 쥐의 귀 염증 억제율을 나타낸 그래프이다.FIG. 7 is a graph showing the inhibition rate of ear inflammation in rats by the disinfectant dithranol when the cacao bean fraction was orally treated in mice.
도 8은 카카오 콩껍질 분획물을 쥐에 경구 처치했을 때, 염증 유발 물질인 디스라놀에 의한 쥐의 귀 염증 억제율을 나타낸 그래프이다.Figure 8 is a graph showing the inhibition rate of ear inflammation in rats by disinol, an inflammation-causing substance, when cacao bean skin fractions were orally treated in rats.
도 9는 항염증 활성을 갖는 카카오 콩 및 콩껍질 분획물들과 식품 중에 존재하는 항염증 물질 중의 하나인 녹차의 EGCG(epigallocatechingallate)를 경구투여시, 피부 염증 유발 물질인 TPA에 의한 쥐의 귀 염증 억제율을 비교한 그래프이다.Figure 9 shows the inhibition rate of ear inflammation in rats by TPA, a skin irritant, when oral administration of cacao beans and bean husk fractions having anti-inflammatory activity and EGCG (epigallocatechingallate) of green tea, which is one of anti-inflammatory substances present in food, This is a graph comparing.
본 발명은 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제를 그 특징으로 한다.The present invention is characterized by an anti-inflammatory food additive containing cacao beans or soybean hull fractions.
이와 같은 본 발명을 상세히 설명하면 다음과 같다.The present invention will be described in detail as follows.
본 발명은 세균이 분비하는 내독소에 의해 발생하는 소화기계 염증질환 및 여러 피부염증 유발 물질들에 의해 발생되는 원발성 접촉피부염에 대해 뛰어난 억제 효능을 확인함으로써, 카카오 콩 또는 콩껍질 분획물을 함유하는 항염증용 식품첨가제에 관한 것이다.The present invention is to determine the excellent inhibitory effect against inflammatory diseases of the digestive system caused by endotoxin secreted by bacteria and primary contact dermatitis caused by various dermatitis-causing substances, thereby containing an anti-cacao bean or soybean peel fraction. It relates to a food additive for inflammation.
본 발명에 따른 카카오 콩으로부터 코코아 버터를 제거하고 남은 분획물(카카오 메스) 또는 카카오 콩껍질 1 중량부에 50% 아세톤 수용액, 50% 에탄올 수용액 또는 50% 메탄올 수용액[Duksan, Korea] 4 ∼ 10 중량부를 첨가하여 40 ∼ 70 ℃의 온도로 4 ∼ 6시간 동안 환류하면서 교반 추출한 다음, 추출물을 원심분리하여 얻은 상징액을 모으고 남은 잔사를 반복 추출한 후, 상기 추출물을 여과, 건조시켜 카카오 콩 또는 콩껍질 추출물을 얻는다. 이 추출물을 스티렌계 다공성 수지에 흡착시킨 후, 20% 에탄올 수용액으로 분획한 후, 다시 60% 에탄올 수용액을 첨가하여 용매 분획물을 얻는다.Remove the cocoa butter from cacao beans according to the present invention in the remaining fractions (cacao) or cacao beans peel 1 part in 50% acetone, 50% ethanol or 50% methanol solution [Duksan, Korea] 4 to 10 parts by weight After stirring and extracting under reflux for 4-6 hours at a temperature of 40-70 ° C., the supernatant obtained by centrifuging the extract was collected and the remaining residue was repeatedly extracted, and then the extract was filtered and dried to extract cacao beans or soybean extract. Get The extract is adsorbed onto a styrene-based porous resin, fractionated with 20% aqueous ethanol solution, and then 60% aqueous ethanol solution is added to obtain a solvent fraction.
상기 분획물을 쥐에게 경구 투여하였을 경우, 소화기계 염증을 유발하기 위하여 세균의 내독소인 LPS를 복강 내에 주사함으로써 발생되는 쥐의 치사율을 현저히 낮추며, 또한 원발성 접촉피부염을 유발하는 물질인 TPA, 디스라놀, 크로톤오일에 의해 발생되는 염증을 현저히 억제시킨다.When the fraction is administered orally to rats, it significantly lowers the mortality rate of rats caused by injection of bacterial endotoxin LPS intraperitoneally to induce digestive system inflammation, and also TPA and dislanol, substances which cause primary contact dermatitis. Inflammation caused by croton oil is significantly suppressed.
한편, 본 발명은 카카오 콩 또는 콩껍질 분획물을 유효성분으로 하는 의약품 또는 식품 첨가제를 포함하는 바, 상기 추출물 의약물 또는 식품첨가제로의 제조방법은 공지방법을 따른다.On the other hand, the present invention comprises a drug or a food additive containing cacao beans or soybean skin fraction as an active ingredient, the method of producing the extract drug or food additives according to a known method.
의약물으로 제조시에는 카카오 콩 또는 콩껍질 분획물 그 자체로도 사용할 수 있지만, 약학적으로 허용되는 담체(carrier), 부형제(forming agent), 희석제(diluent) 등과 혼합하여 분말, 과립, 캡슐 또는 주사제 등으로도 제조가 가능하다. 또한, 본 발명에 따른 카카오 콩은 예로부터 식용으로 사용되어온 것으로 분획물의 투여용량에 특별한 제약은 없고, 체내 흡수도, 체중, 환자의 연령,성별, 건강상태 식이, 투여시간, 투여 방법, 배설율, 질환의 중증도 등에 따라 변화될 수 있다. 일반적으로 카카오 콩 또는 콩껍질 분획물은 체중 1 ㎏당 0.1 내지 10 ㎎ 정도를 투여하는 것이 바람직하다. 따라서, 본 발명의 유효성분을 포함하는 약제는 유효량 범위를 고려하여 제조하도록 하며, 이렇게 제형화된 단위투여형 제제는 필요에 따라 약제의 투여를 감시하거나 관찰하는 전문가의 판단과 개인의 요구에 따라 전문화된 투약법을 사용하거나 일정시간 간격으로 수회 투여할 수 있다.When prepared as a pharmaceutical, cacao beans or bean husk fractions may be used as such, but may be mixed with pharmaceutically acceptable carriers, forming agents, diluents, etc., to give powders, granules, capsules or injectables. It is also possible to manufacture. In addition, cacao beans according to the present invention has been used for edible since ancient times, there is no particular restriction on the dosage of the fraction, body absorption, weight, age of the patient, sex, health status, administration time, administration method, excretion rate And the severity of the disease. In general, cacao beans or bean husk fractions are preferably administered at about 0.1 to 10 mg per kg body weight. Therefore, the drug containing the active ingredient of the present invention is to be prepared in consideration of the effective amount range, and the unit dosage form formulated in this way according to the judgment of experts and the needs of the individual to monitor or observe the administration of the drug as needed Specialized medications can be used or administered at regular intervals.
한편, 식품첨가제로 제조시에는 음료, 껌, 과자류 등의 식품소재에 본 발명의 카카오 콩 또는 콩껍질 분획물을 포함시켜 제조할 수 있다.On the other hand, when manufacturing as a food additive may be prepared by including the cacao beans or bean husk fraction of the present invention in food materials such as beverages, gum, confectionery.
이상에서 설명한 카카오 콩 또는 콩껍질 분획물을 유효성분으로 함유하는 의약품 및 식품첨가제는 염증 예방 및 억제효과에 탁월하다.Drugs and food additives containing the cacao beans or soybean peel fraction as described as an active ingredient is excellent for the prevention and inhibition of inflammation.
이하, 본 발명을 실시예에 의거하여 더욱 상세히 설명하겠는바, 본 발명이 다음 실시예로 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited to the following Examples.
실시예 1: 카카오 콩으로부터 염증 억제 효능을 갖는 분획물의 제조Example 1: Preparation of Fractions Having Inhibitory Effect from Cacao Beans
<1-1> 용매 추출에 의한 염증 억제 효능을 갖는 추출물의 제조<1-1> Preparation of Extracts Having Inhibitory Effect by Solvent Extraction
카카오 콩으로부터 코코아 버터를 제거하고 남은 분획물(카카오 메스) 1 중량부에 50% 아세톤[Duksan, Korea] 수용액 6 중량부를 첨가하여 60 ℃의 온도로 5시간 동안 환류하면서 교반 추출한 후, 이 추출물은 8,000 rpm, 4 ℃, 30분간 원심분리[Vision, Korea]하여 얻어진 상징액을 모으고, 남은 잔사는 동일한 방법으로 1회 반복하여 추출하였다. 이렇게 얻어진 추출물을 여과지(Whatmann 41)로 여과시킨 후, 냉동 건조하여 카카오 콩 추출물을 얻었다.After removing cocoa butter from cacao beans, 6 parts by weight of 50% acetone [Duksan, Korea] aqueous solution was added to 1 part by weight of the remaining fraction (cacao), and the mixture was stirred and refluxed at a temperature of 60 ° C. for 5 hours. The supernatant obtained by centrifugation [Vision, Korea] for 30 minutes at rpm, 4 ° C. was collected, and the remaining residue was repeatedly extracted in the same manner. The extract thus obtained was filtered through filter paper (Whatmann 41), and then freeze-dried to obtain a cacao bean extract.
<1-2> 흡착 칼럼 크로마토그래피에 의한 염증 억제 효능을 갖는 분획물의 제조<1-2> Preparation of Fractions Having Inhibition of Inflammation by Adsorption Column Chromatography
상기 <1-1>에서 얻어진 추출물로부터 기존에 특허 출원한 방법을 참고로 하여[미국 특허 제 6159451호, 2000] 흡착 칼럼크로마토그래피에 의해 분획물을 제조하였다. 즉, 소수성 충진제인 XAD-4[Sigma, USA]나 HP-20[Mitsbish, Japan]에 흡착시킨 후 20% 에탄올로 세척하고, 남은 분획을 다시 60% 에탄올로 분획하였다.From the extract obtained in the above <1-1>, a fraction was prepared by adsorption column chromatography with reference to a previously patented method [US Pat. No. 6159451, 2000]. That is, it was adsorbed on the hydrophobic filler XAD-4 [Sigma, USA] or HP-20 [Mitsbish, Japan], washed with 20% ethanol, and the remaining fractions were fractionated with 60% ethanol again.
실시예 2 : 카카오 콩껍질로부터 염증 효능을 갖는 분획물의 제조Example 2 Preparation of Fractions Having Inflammatory Effect from Cacao Bean Peel
<2-1> 용매 추출에 의한 카카오 콩껍질로부터 염증 억제 효능을 갖는 추출물의 제조<2-1> Preparation of an extract having inhibitory effect from cacao bean husk by solvent extraction
카카오 콩껍질 1 중량부에 50% 아세톤[Duksan, Korea] 수용액 6 중량부를 첨가하여 60 ℃의 온도로 5시간 동안 환류하면서 교반 추출한 후, 이 추출물은 8,000 rpm, 4 ℃, 30분간 원심분리[Vision, Korea]하여 얻어진 상징액을 모으고, 남은 잔사는 동일한 방법으로 1회 반복하여 추출하였다. 이렇게 얻어진 추출물을 여과지(Whatmann 41)로 여과시킨 후, 냉동 건조하여 카카오 콩껍질 추출물을 얻었다.6 parts by weight of 50% acetone [Duksan, Korea] aqueous solution was added to 1 part by weight of cacao beans, and the mixture was stirred under reflux for 5 hours at a temperature of 60 ° C., and then the extract was centrifuged at 8,000 rpm, 4 ° C. for 30 minutes [Vision , Korea], the supernatant obtained was collected, and the remaining residue was repeatedly extracted once in the same manner. The extract thus obtained was filtered through filter paper (Whatmann 41), and then freeze-dried to obtain a cacao bean peel extract.
<2-2> 흡착 칼럼 크로마토그래피에 의한 염증 억제 효능을 갖는 분획물의 제조<2-2> Preparation of Fractions Having Inhibitory Effect of Inflammation by Adsorption Column Chromatography
상기 <2-1>에서 얻어진 추출물로부터 기존에 특허 출원한 방법을 참고로 하여[미국 특허 제6159451호, 2000] 흡착 칼럼크로마토그래피에 의해 분획물을 제조하였다. 즉, 소수성 충진제인 XAD-4[Sigma, USA]나 HP-20[Mitsbish, Japan]에 흡착시킨 후 20% 에탄올로 세척하고, 남은 분획을 다시 60% 에탄올로 분획하였다.Fractions were prepared by adsorption column chromatography from the extract obtained in the above <2-1> with reference to a method of patent application [US Patent No. 6159451, 2000]. That is, it was adsorbed on the hydrophobic filler XAD-4 [Sigma, USA] or HP-20 [Mitsbish, Japan], washed with 20% ethanol, and the remaining fractions were fractionated with 60% ethanol again.
상기와 같이 얻어진 카카오 콩 또는 콩껍질 분획물의 항염증 효능을 평가하기 위한 다음과 같은 실험들을 실시하였다.The following experiments were carried out to evaluate the anti-inflammatory efficacy of the cacao beans or soy bark fraction obtained as described above.
실시예 3: 세균의 내독소에 의해 발생되는 염증의 억제 활성Example 3: Inhibitory Activity of Inflammation Caused by Bacterial Endotoxin
상기 실시예 1, 2에서 얻은 분획물들의 세균의 내독소에 의해 발생하는 염증에 대한 억제 활성은 다음 공지의 방법[Yang et al., J. Nutr., 128: 2334-2340, 1998]에 따라 실시하였다.The inhibitory activity against inflammation caused by bacterial endotoxin of the fractions obtained in Examples 1 and 2 was carried out according to the following known method [Yang et al., J. Nutr., 128: 2334-2340, 1998]. It was.
우선, 25 ∼ 30 g의 BALB/C 쥐를 한 군당 25 마리를 준비하여 실험 전에 12시간 절식시킨 후, 상기의 방법에 의해 얻어진 시료를 1 g/㎏ 농도로 쥐에 경구 투여시켰다. 반면에 대조군은 동량의 인산용액만을 경구투여시켰다. 30분 후에 자이라진 하이드로클로라이드(xylazine hydrochloride)로 마취시킨 후 다시 5분 후에 40 ㎎/㎏의 LPS를 복강내로 투여하였다. 30 시간까지 매 2시간마다 생존율을 측정하였다. 항염증 정도는 대조군에 대한 생존 수의 차이로 측정하였다.First, 25 to 30 g of BALB / C mice were prepared per group and fasted for 12 hours before the experiment, and the samples obtained by the above method were orally administered to the mice at a concentration of 1 g / kg. On the other hand, the control group was orally administered only the same amount of phosphate solution. After 30 minutes of anesthesia with xylazine hydrochloride, after 5 minutes, 40 mg / kg of LPS was intraperitoneally administered. Survival was measured every 2 hours until 30 hours. The degree of anti-inflammatory was measured by the difference in survival numbers for the control group.
상기 카카오 콩 및 콩껍질 분획물들은 모두 세균의 내독소이면서 염증유발 물질인 LPS에 의해 발생하는 쥐의 치사율에 대한 저해 효능을 실험한 결과를 도 1과 2에 나타냈다. 도 1, 2에 나타낸 바와 같이, 카카오 콩 및 콩껍질 분획물을 각각 1 g/㎏ 농도로 쥐에 경구 투여시 30 시간에서 각각 32%와 36%의 치사율 감소가 관찰되었다. 특히, 12 시간에서는 각각 44%과 48%의 높은 치사율 감소 효과를 나타내었다.The cacao beans and bean husk fractions are shown in Figures 1 and 2 to test the inhibitory effect on the mortality of mice caused by LPS, which is a bacterial endotoxin and inflammation-inducing substance. As shown in Figures 1 and 2, the mortality decreases of 32% and 36% were observed at 30 h, respectively, when orally administered cacao beans and soybean skin fractions to mice at a concentration of 1 g / kg respectively. In particular, at 12 hours, the mortality rate was 44% and 48%, respectively.
실시예 4: TPA에 의해 유발되는 염증의 억제 활성Example 4: Inhibitory Activity of Inflammation Induced by TPA
상기 실시예 1, 2에서 얻은 분획물들의 TPA에 의해 발생하는 염증의 억제 활성은 다음 공지의 방법[Romay C., et al., Inflamm. Res. 47: 334-338, 1998]에 따라 실시하였다.Inhibitory activity of inflammation caused by TPA of the fractions obtained in Examples 1 and 2 was determined by the following known methods [Romay C., et al., Inflamm. Res. 47: 334-338, 1998].
우선, 25 ∼ 30 g의 BALB/C 쥐를 한 군당 8 마리를 준비하여 실험 전에 12시간 절식시킨 후, 상기의 방법에 의해 얻어진 시료를 각각의 농도로 경구 투여시켰다. 30분 후에 자이라진 하이드로클로라이드로 마취시킨 후 다시 5분 후에 TPA를 귀당 25 ㎍ 씩 양쪽 귀에 도포하였다. 5시간 후에 오른쪽 및 왼쪽 귀를 6 mm 펀치로 잘라내어 무게를 측정하였다. 항염증 정도는 대조군에 대한 무게의 차이로 측정하였다. (도 3 내지 도 9는 평균값으로 나타냈으며, 통계적 유의성은 유의수준 * P < 0.05, ** P < 0.01 에서 two-tailed Dunnett's t-test를 따라서 ANOVA 법[Ott, R. L., An introduction to statistical methods and data analysis, 1997]에 의해 검정하였다.)First, 8 rats per group of 25-30 g BALB / C mice were prepared and fasted 12 hours before the experiment, and the samples obtained by the above method were orally administered at respective concentrations. After 30 minutes of anesthesia with xyrazin hydrochloride, another 5 minutes later TPA was applied to both ears at 25 μg per ear. After 5 hours the right and left ears were cut out with a 6 mm punch and weighed. The degree of anti-inflammatory was measured by the difference in weight with respect to the control. (Figs. 3 to 9 are shown as averages, and statistical significance is determined by the ANOVA method [Ott, RL, An introduction to statistical methods and according to the two-tailed Dunnett's t-test at the significance level * P <0.05, ** P <0.01. data analysis, 1997].
상기 카카오 콩 및 콩껍질 분획물들의 염증유발 물질인 TPA에 의해 발생하는 염증에 대한 저해 효능을 실험한 결과를 도 3과 4에 나타냈다. 도 3, 4에 나타낸 바와 같이, 카카오 콩 및 콩껍질 분획물들을 각각 0.5 g/㎏ 농도로 쥐에 경구 투여시 62%와 64%의 염증 억제율을 나타내었으며, 1 g/㎏ 농도로 쥐에 경구 투여시 각각 72%과 68%의 염증 억제율을 나타냄으로써 유사한 저해 효능을 나타내었다.3 and 4 show the results of experiments on the inhibitory effect on inflammation caused by TPA, which is an inflammation-causing substance of cacao beans and bean husk fractions. As shown in Figures 3 and 4, the cacao bean and soybean hull fractions showed an inhibition rate of 62% and 64% when administered orally to rats at a concentration of 0.5 g / kg, respectively, and administered orally to mice at a concentration of 1 g / kg. Inhibition rate of 72% and 68%, respectively, showed similar inhibitory efficacy.
실시예 5: 크로톤 오일에 의해 발생하는 염증의 억제 활성Example 5 Inhibitory Activity of Inflammation Caused by Croton Oil
상기 실시예 1, 2에서 얻은 분획물들의 크로톤 오일에 의해 발생하는 염증의 억제 활성은 다음 공지의 방법[Clementi G., et al., Life Sci. 57: 193-197, 1995; Heiman A. S., et al., Steroids 62: 491-499, 1997]에 따라 실시하였다.The inhibitory activity of inflammation caused by croton oil of the fractions obtained in Examples 1 and 2 was determined by the following known method [Clementi G., et al., Life Sci. 57: 193-197, 1995; Heiman A. S., et al., Steroids 62: 491-499, 1997.
상기의 방법과 유사하게, 시료들을 각각의 농도로 경구 투여시켰다. 30분 후에 자이라진 하이드로클로라이드로 마취시킨 후 다시 5분 후에 크로톤 오일을 귀당 20 ㎕ 씩 양쪽 귀에 도포하였다. 5시간 후에 오른쪽 및 왼쪽 귀를 6 mm 펀치로 잘라내어 무게를 측정하였다. 항염증 정도는 대조군에 대한 무게의 차이로 측정하였다.Similar to the above method, samples were administered orally at respective concentrations. After 30 minutes of anesthesia with xyrazin hydrochloride, again 5 minutes later croton oil was applied to both ears at 20 μl per ear. After 5 hours the right and left ears were cut out with a 6 mm punch and weighed. The degree of anti-inflammatory was measured by the difference in weight with respect to the control.
상기 카카오 콩 및 콩껍질 분획물들의 염증유발 물질인 크로톤 오일에 의해 발생하는 염증에 대한 억제 효능을 실험한 결과를 도 5와 6에 나타냈다. 도 5, 6에 나타낸 바와 같이, 카카오 콩 및 콩껍질 분획물들을 각각 0.5 g/㎏ 농도로 쥐에 경구 투여시 67%와 53%의 염증 억제율을 나타내었으며, 1 g/㎏ 농도로 쥐에 경구 투여시 각각 77%과 71%의 염증 억제율을 나타냄으로써 유사한 억제 효능을 나타내었다. 도 5와 6에 나타낸 바와 같이, 카카오 콩 및 콩껍질 분획물들을 각각 1 g/㎏ 농도로 쥐에 경구 투여시 77%와 68%의 강한 염증 억제 활성을 나타내었다.5 and 6 show the results of experiments on the inhibitory effect of inflammation caused by croton oil, an inflammation-causing substance of cacao beans and soybean hull fractions. As shown in Figures 5 and 6, the cacao bean and soybean hull fractions showed an inhibitory rate of 67% and 53% when administered orally to rats at a concentration of 0.5 g / kg, respectively, and orally to rats at a concentration of 1 g / kg. Similar inhibitory efficacy was shown by showing inflammatory inhibition rates of 77% and 71%, respectively. As shown in Figures 5 and 6, cacao beans and soy bark fractions showed a strong inflammatory inhibitory activity of 77% and 68% when orally administered to mice at a concentration of 1 g / kg, respectively.
실시예 6: 디스라놀에 의해 발생하는 염증의 억제 활성Example 6: Inhibitory Activity of Inflammation Caused by Disranol
상기 실시예 1, 2에서 얻은 분획물들의 디스라놀에 의해 발생하는 염증의 억제 활성은 다음 공지의 방법[Viluksela M., et al., Arch. Dermatol. Res. 283: 262-268, 1991; Blazso G., et al., Prostaglandins 50: 161-168, 1995]에 따라 실시하였다.The inhibitory activity of inflammation caused by dislanol of the fractions obtained in Examples 1 and 2 was determined by the following known methods [Viluksela M., et al., Arch. Dermatol. Res. 283: 262-268, 1991; Blazso G., et al., Prostaglandins 50: 161-168, 1995].
상기의 방법과 유사하게, 시료들을 각각의 농도로 경구 투여시켰다. 30분 후에 자이라진 하이드로클로라이드로 마취시킨 후 다시 5분 후에 디스라놀을 귀당 50 ㎍씩 양쪽 귀에 도포하였다. 5시간 후에 오른쪽 및 왼쪽 귀를 6 mm 펀치로 잘라내어 무게를 측정하였다. 항염증 정도는 대조군에 대한 무게의 차이로 측정한다.Similar to the above method, samples were administered orally at respective concentrations. After 30 minutes of anesthesia with xyrazin hydrochloride, another 5 minutes later dislanol was applied to both ears at 50 μg per ear. After 5 hours the right and left ears were cut out with a 6 mm punch and weighed. The degree of anti-inflammatory is measured by the difference in weight relative to the control.
상기 카카오 콩 및 콩껍질 분획물들의 염증유발 물질인 디스라놀에 의해 발생하는 염증에 대한 억제 효능을 실험한 결과를 도 7과 8에 나타냈다. 도 7과 8에 나타낸 바와 같이, 카카오 콩 및 콩껍질 분획물들을 각각 1 g/㎏ 농도로 쥐에 경구 투여시 82%과 78%의 강한 염증 억제 활성을 나타내었고, 또한 0.5 g/㎏ 농도에서 각각 51%과 59%의 염증 억제율을 나타내었다.7 and 8 show the results of experiments on the inhibitory effect on the inflammation caused by dislanol, an inflammation-causing substance, of the cacao beans and bean husk fractions. As shown in Figs. 7 and 8, cacao bean and soybean skin fractions showed strong inflammatory inhibitory activity of 82% and 78% when administered orally to rats at a concentration of 1 g / kg, respectively, and at a concentration of 0.5 g / kg, respectively. Inflammation inhibition rates were 51% and 59%.
이상과 같이, 상기 카카오 콩 및 콩껍질 분획물들을 경구 투여시 TPA, 크로톤 오일, 디스라놀에 의해 유발되는 염증의 강한 저해 활성을 나타냈다. 또한, 처리한 시료의 농도가 점차 증가할수록 염증이 억제되는 것을 관찰함으로써, 염증들이 경구 투여한 카카오 콩 및 콩껍질 분획물들의 농도에 의존적인 것을 알 수 있었다.As described above, oral administration of the cacao beans and bean husk fractions showed a strong inhibitory activity of inflammation caused by TPA, croton oil, dislanol. In addition, by observing that inflammation is inhibited as the concentration of the treated sample is gradually increased, it can be seen that the inflammation is dependent on the concentration of cacao beans and soybean skin fractions administered orally.
상기의 실험 결과, 카카오 콩 및 콩껍질의 분획물들을 쥐에게 경구 투여하였을 때 강력한 염증 억제 효능을 나타내었다.As a result of the above experiment, the fractions of cacao beans and soybean hulls showed strong anti-inflammatory effect when orally administered to rats.
실시예 7: 정제의 제조Example 7: Preparation of Tablets
유효성분 10 g10 g of active ingredients
락토스 70 g70 g of lactose
결정성 셀룰로오스 15 g15 g of crystalline cellulose
마그네슘 스테아레이트 5 g5 g of magnesium stearate
총 량 100 gTotal amount 100 g
상기에서 나열된 성분들을 잘게 부숴 혼합한 후 직타법(direct tableting method)에 의해 정제를 제조하였다. 각 정제의 총량은 100 ㎎이고, 그 중 유효성분의 함량은 10 ㎎이다.The tablets were prepared by direct tableting method after mixing the ingredients listed above finely. The total amount of each tablet is 100 mg, of which the active ingredient content is 10 mg.
실시예 8: 분말제의 제조Example 8: Preparation of Powder
유효성분 10 g10 g of active ingredients
옥수수 전분 50 g50 g of corn starch
카르복시 셀룰로오스 40 g40 g of carboxy cellulose
총 량 100 gTotal amount 100 g
상기에서 나열된 성분들을 잘게 부숴 혼합하여 분말을 제조하였다. 경질캡슐에 분말 100 ㎎을 넣어 캡슐제를 제조하였다.A powder was prepared by crushing and mixing the ingredients listed above. 100 mg of powder was put into the hard capsule to prepare a capsule.
실시예 9: 독성 시험Example 9: Toxicity Test
본 발명의 카카오 콩 또는 콩껍질 분획물에 대하여 독성실험을 다음과 같이 수행하였다. 구체적으로 상기 분획물을 디메틸설폭사이드(dimethylsulfoxide, 이하 DMSO)에 용해하고 물로 희석한 후 이를 마우스(군당 10마리)에 각각 1 g/㎏을 투여한 다음 7일간 관찰하였으나 사망하는 쥐는 없었다.Toxicity experiments were performed on cacao beans or bean hull fractions of the present invention as follows. Specifically, the fractions were dissolved in dimethylsulfoxide (DMSO), diluted with water, and then administered to each mouse (10 mice per group) 1 g / kg and observed for 7 days, but no rats died.
실시예 10Example 10
껌 베이스 20 중량%, 설탕 76.9 중량%, 향료 1 중량% 및 물 2 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 0.1 중량%를 배합하여 통상의 방법으로 츄잉껌을 제조하였다.Chewing gum was prepared in a conventional manner by combining 20% by weight of gum base, 76.9% by weight of sugar, 1% by weight of perfume, and 2% by weight of water, and 0.1% by weight of cacao beans or legume fractions obtained in the above examples.
실시예 11Example 11
설탕 60 중량%, 물엿 39.8 중량% 및 향료 0.1 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 0.1 중량%를 배합하여 통상의 방법으로 캔디를 제조하였다.Candy was prepared in a conventional manner by combining 60% by weight of sugar, 39.8% by weight of starch syrup, and 0.1% by weight of fragrance, and 0.1% by weight of cacao beans or bean husk fraction obtained in the above examples.
실시예 12Example 12
박력 1급 25.59 중량%, 중력 1급 22.22 중량%, 정백당 4.80 중량%, 식염0.73 중량%, 포도당 0.78 중량%, 팜쇼트닝 11.78 중량%, 암모 1.54 중량%, 중조0.17 중량%, 중아황산나트륨 0.16 중량%, 쌀가루 1.45 중량%, 비타민 B₁0.0001 중량%, 비타민 B₂ 0.0001 중량%, 밀크향 0.04 중량%, 물 20.6998 중량%, 전지분유 1.16 중량% 대용분유 0.29 중량%, 제일인산칼슘 0.03 중량%, 살포염 0.29 중량% 및 분무유 7.27 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 1중량%를 배합하여 통상의 방법으로 비스켓을 제조하였다.Force 1st grade 25.59 wt%, gravity 1st grade 22.22 wt%, white sugar 4.80 wt%, salt 0.73 wt%, glucose 0.78 wt%, palm shortening 11.78 wt%, ammo 1.54 wt%, sodium bicarbonate 0.17 wt%, sodium bisulfite 0.16 wt% , Rice flour 1.45 wt%, Vitamin B₁0.0001 wt%, Vitamin B₂ 0.0001 wt%, Milk flavor 0.04 wt%, Water 20.6998 wt%, Whole milk powder 1.16 wt% Substitute powder 0.29 wt%, calcium phosphate 0.03 wt%, Spray salt Biscuits were prepared in a conventional manner by combining 0.29% by weight and 7.27% by weight of spray oil with 1% by weight of cacao beans or legume fractions obtained in the above examples.
실시예 13Example 13
꿀 0.26 중량%, 치옥토산아미드 0.0002 중량%, 니코틴산아미드 0.0004 중량%, 염산리보플라빈나트륨 0.0001 중량%, 염산피리독신 0.0001 중량%, 이노시톨 0.001 중량%, 오르트산 0.002 중량% 및 물 98.7362 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 1 중량%를 배합하여 통상의 방법으로 음료를 제조하였다.0.26% by weight of honey, 0.0002% by weight of thioctoamide, 0.0004% by weight of nicotinic acid, 0.0001% by weight of riboflavin hydrochloride, 0.0001% by weight of pyridoxine hydrochloride, 0.001% by weight of inositol, 0.002% by weight of orthoic acid and 98.7362% by weight of water The beverage was prepared by the conventional method by combining the cacao beans or bean husk fraction obtained in the example.
실시예 14Example 14
돈육 65.18 중량%, 계육 25 중량%, 전분 3.5 중량%, 대두단백 1.7 중량%, 식염 1.62 중량%, 포도당 0.5 중량% 및 글리세린 1.5 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 1 중량%를 배합하여 통상의 방법으로 소세지를 제조하였다.65.18% pork, 25% poultry, 3.5% starch, 1.7% soy protein, 1.62% salt, 0.5% glucose and 1.5% glycerin and 1% cacao beans or bean husk fractions obtained in the above examples. Sausage was prepared by combining the conventional method.
실시예 15Example 15
스피루리나 55 중량%, 구아검효소 분해물 10 중량%, 비타민 B₁염산염 0.01중량%, 비타민 B6 염산염 0.01 중량%, DL-메티오닌 0.23 중량%, 스테아린산 마그네슘 0.7 중량%, 유당 22.2 중량% 및 옥수수전분 1.85 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 10 중량%를 배합하여 통상의 방법으로 정제형 건강보조식품을 제조하였다55% by weight of spirulina, 10% by weight of guar gum enzyme, 0.01% by weight of vitamin B ₁ hydrochloride, 0.01% by weight of vitamin B6 hydrochloride, 0.23% by weight of DL-methionine, 0.7% by weight of magnesium stearate, 22.2% by weight of lactose and 1.85% by weight of corn starch And 10% by weight of cacao beans or bean husk fraction obtained in the above Example was prepared a tablet-type health supplement in a conventional manner.
실시예 16Example 16
키토올리고당 11.26 중량%, 마늘분말 0.2 중량%, 은행잎추출물 분말 0.2 중량%, 베타카로틴(30% 현탄액) 0.9 중량%, 알파토코페롤 1.2 중량%, 레시틴 1.2 중량%, 정제팜유 4.5 중량%, 황납 1.6 중량%, 대두유 18.994 중량%, 젤라틴 37.83 중량%, 글리세린 16.51 중량%, 에틸바닐린 0.09 중량% 이산화티타늄 0.076 중량% 및 식용색소 0.44 중량%와 상기 실시예에서 얻은 카카오 콩 또는 콩껍질 분획물 5 중량%를 배합하여 통상의 방법으로 캡슐형 건강보조식품을 제조하였다.Chitooligosaccharide 11.26 wt%, Garlic powder 0.2 wt%, Ginkgo biloba extract powder 0.2 wt%, Beta-carotene (30% suspension) 0.9 wt%, Alpha tocopherol 1.2 wt%, Lecithin 1.2 wt%, Refined palm oil 4.5 wt%, Hest 1.6 Weight%, soybean oil 18.994 weight%, gelatin 37.83 weight%, glycerin 16.51 weight%, ethyl vanillin 0.09 weight% titanium dioxide and 0.076 weight% food coloring and 0.44 weight% cacao beans or soybean skin fractions obtained in the above examples. By blending to prepare a capsule-type dietary supplement in a conventional manner.
비교예Comparative example
이 분획의 활성을 일반적으로 잘 알려진 항염증 물질인 녹차의 EGCG(epigallocatechingallate)와 비교하기 위하여 상기 실시예 4와 동일한 방법으로 실험하였다. 도 9에서 보듯이, 카카오 콩 및 콩껍질의 분획물은 순수한 항염증 물질인 EGCG와 비교했을 때 매우 우수한 효능을 나타내었다.In order to compare the activity of this fraction with EGCG (epigallocatechingallate) of green tea, which is a generally well known anti-inflammatory substance, it was tested in the same manner as in Example 4. As shown in Figure 9, the fractions of cacao beans and soybean hulls showed very good efficacy compared to EGCG, a pure anti-inflammatory material.
결국, 본 발명에서 사용한 방법에 의한 카카오 콩 및 콩껍질 분획물은 동물 실험에서 강한 항염증 활성을 갖는 것을 알 수 있었다.As a result, it was found that cacao beans and legume fractions by the method used in the present invention have strong anti-inflammatory activity in animal experiments.
상기에서 살펴본 바와 같이, 본 발명에 따른 카카오 콩 또는 콩껍질 분획물은 동물 실험 결과, 경구 투여시 염증에 대한 억제 효능이 우수하므로 이를 염증 억제제로 사용하면, 세균의 내독소 및 외부 염증 유발물질에 의해 발생되는 염증을 효과적으로 억제할 수 있을 뿐만 아니라, 식품 산업에 이용 가능한 용매 추출 및 간단한 흡착 칼럼크로마토그래피를 이용하여 항염증 활성 분획만을 제제화하면 되므로, 값싸게 가공식품에 이용할 수 있으므로 기존의 항염증 제제에 비하여 생산 원가를 낮출 수 있으며, 특히 막대한 양의 카카오 콩껍질을 재활용함으로써 경제성을 향상시킬 수 있다.As described above, the cacao beans or soybean peel fraction according to the present invention has an excellent inhibitory effect on inflammation when administered orally, when used as an inhibitor of inflammation, by bacterial endotoxin and external inflammatory agents In addition to effectively suppressing the inflammation that occurs, only anti-inflammatory active fractions can be formulated using solvent extraction and simple adsorption column chromatography, which can be used in the food industry. Compared with this, the production cost can be lowered, and the economic efficiency can be improved by recycling large amounts of cacao beans.
아울러, 식품 등급으로 사용되는 용매 및 칼럼크로마토그래피를 사용하므로, 독성을 제거하기 위한 별도의 정제 과정 없이도 안전하게 섭취할 수 있다. 따라서, 상기 카카오 콩 또는 콩껍질 분획물을 첨가하여 가공식품을 생산할 수 있으며, 이러한 가공식품의 섭취에 의해 염증을 억제하는 효능을 기대할 수 있다.In addition, since the solvent and column chromatography used as food grade, it can be safely ingested without a separate purification process to remove toxicity. Therefore, it is possible to produce processed foods by adding the cacao beans or bean husk fraction, it can be expected to have the effect of inhibiting inflammation by the ingestion of such processed foods.
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