KR20210043806A - Composition for Anti-Obesity Comprising Catechin, and Complex Extracts of Brown Rices, Black Soybean, and Yeast as Active Ingredient - Google Patents
Composition for Anti-Obesity Comprising Catechin, and Complex Extracts of Brown Rices, Black Soybean, and Yeast as Active Ingredient Download PDFInfo
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- KR20210043806A KR20210043806A KR1020190126579A KR20190126579A KR20210043806A KR 20210043806 A KR20210043806 A KR 20210043806A KR 1020190126579 A KR1020190126579 A KR 1020190126579A KR 20190126579 A KR20190126579 A KR 20190126579A KR 20210043806 A KR20210043806 A KR 20210043806A
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- catechin
- obesity
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- yeast
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Abstract
Description
본 발명은 카테킨과 현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는 항비만용 조성물에 관한 것이다.The present invention relates to a composition for anti-obesity comprising a complex extract of catechin and brown rice, seomoktae, and yeast as an active ingredient.
체지방이 건강에 해로운 영향을 미칠 정도로 과도하게 축적된 상태를 비만이라고 한다. 음식으로 섭취한 열량이 신체활동으로 소비하는 열량보다 많을 때 체지방으로 축적되는데, 남자는 체지방이 체중의 25% 이상일 때, 여자는 체중의 30% 이상일 때를 비만이라고 정의한다.Obesity is a condition in which body fat is accumulated excessively enough to have a detrimental effect on health. When the amount of calories consumed by food is greater than the amount of calories consumed by physical activity, it is accumulated as body fat. When the body fat is 25% or more of the body weight in men, and when the woman is 30% or more of the body weight, it is defined as obesity.
비만의 원인은 시상하부의 기능 및 에너지 대사의 이상과 같은 질환과 유전적인 요인이 있으나, 대부분의 비만은 과도한 영양섭취와 신체 활동량의 감소로 인한 생활습관으로 발생한다. 최근에는 서구적인 식생활 패턴과 생활의 편리화로 인하여 즉석식품의 섭취가 증가하고, 운동 부족 현상이 증가하여 비만 발병률이 지속해서 증가하는 추세이며, 이러한 추세가 시간이 지날수록 더욱 심해질 것으로 전망되고 있다.Obesity is caused by diseases such as abnormal hypothalamic function and energy metabolism and genetic factors, but most obesity is caused by excessive nutritional intake and lifestyle due to decrease in physical activity. In recent years, the intake of ready-to-eat foods increases due to the Western dietary pattern and convenience of life, and the incidence of obesity continues to increase due to an increase in lack of exercise, and this trend is expected to become more severe as time passes.
비만의 최대 문제는 단순 비만 그 자체보다는, 비만 상태가 오랜 시간 계속되면서 발생하는 당뇨병, 고지혈증, 심장병, 뇌졸중, 동맥경화증, 지방간 등의 각종 대사성 질환이 문제가 된다. 또한, 비만은 불임 및 월경불순, 퇴행성 관절염, 일부 암, 수면 무호흡, 호흡기 장애, 담석증, 우울증 등 여러 가지 질병의 위험도를 증가시켜 성인병 발생의 위험요소이자 사회문제이다.The biggest problem of obesity is not simple obesity itself, but various metabolic diseases such as diabetes, hyperlipidemia, heart disease, stroke, arteriosclerosis, fatty liver, etc., which occur as obesity continues for a long time. In addition, obesity increases the risk of various diseases such as infertility and menstrual irregularities, degenerative arthritis, some cancers, sleep apnea, respiratory disorders, cholelithiasis, and depression, which is a risk factor and social problem of adult diseases.
최근 사회적으로 비만을 질병으로 인식함으로써, 다양한 제약 회사에서 비만을 치료하는 약물 개발에 박차를 가하고 있다. 지방흡수 저해제인 제니칼은 세계적으로 가장 많이 사용되는 비만 치료제 중 하나이다. 제니칼의 성분인 오르리스타트(orlistat)는 소화된 지방과 결합해 장내에서의 흡수를 억제해 식사 중 지방 성분의 일부를 그대로 배설시키는 작용을 한다. 그밖에 개발된 약물로는 포만감을 증진시키는 리덕틸이나, 엑소리제 등이 있다.Recently, by socially recognizing obesity as a disease, various pharmaceutical companies are spurring the development of drugs to treat obesity. Xenical, an inhibitor of liposuction, is one of the most widely used obesity treatments in the world. Orlistat, an ingredient of Xenical, binds to digested fat and inhibits absorption in the intestine, thereby acting to excrete some of the fat components during the meal as they are. Other drugs developed include reductil, which improves satiety, and exorase.
그러나 상기 상업화된 약물들을 장기간 복용하는 경우, 간 손상, 위장 출혈, 췌장염, 신장 결석을 비롯한 부작용들이 보고되고 있으며, 심장질환, 호흡기 질환, 신경계 질환을 발생할 위험이 있는 것으로 알려져 있다. However, when taking the commercialized drugs for a long time, side effects such as liver damage, gastrointestinal bleeding, pancreatitis, and kidney stones have been reported, and it is known that there is a risk of developing heart disease, respiratory disease, and nervous system disease.
따라서, 현재 상업화된 비만 치료제를 대체하여, 부작용이 적으면서도 비만 개선 효과가 우수한 천연소재의 의약품의 개발이 요구되고 있는 실정이다.Accordingly, there is a demand for the development of a natural drug that has a low side effect and excellent obesity improvement effect by replacing the currently commercialized obesity treatment.
한편, 카테킨은 플라보노이드 그룹의 [flavan-3-ols]에 속하며, 폴리페놀 일종으로 녹차의 떫은맛 성분이다. 차류(Camellia sinensis)에서 파생되었으며, 백차, 녹차, 홍차, 우롱차에 포함되어 있다. 카테킨은 발암 억제, 동맥경화, 혈압상승 억제, 혈전 예방, 항바이러스, 항비만, 항당뇨, 항균, 해독작용, 소염작용, 충치 예방, 구갈 방지, 장내 세균총 정상화 등 다양한 효과가 있는 것으로 알려져 있다. 카테킨의 항비만 활성과 관련하여, 특허등록 제10-1155079호, 및 특허등록 제10-1390366호 등이 카테킨에 대한 항비만 활성을 개시하고 있으나, 그 효과가 미진하여 아직까지 항비만 소재로 카테킨 성분의 현장에서의 활용은 저조한 상황이다.On the other hand, catechin belongs to [flavan-3-ols] of the flavonoid group, and is a kind of polyphenol, which is an astringent component of green tea. Derived from the tea class (Camellia sinensis), it is included in white tea, green tea, black tea, and oolong tea. Catechins are known to have various effects, such as inhibiting carcinogenesis, curing arteries, inhibiting blood pressure rise, preventing blood clots, antiviral, anti-obesity, anti-diabetes, antibacterial, detoxification, anti-inflammatory action, preventing tooth decay, preventing mouth mouth, and normalizing the intestinal flora. Regarding the anti-obesity activity of catechins, Patent Registration No. 10-1155079 and Patent Registration No. 10-1390366 disclose anti-obesity activity against catechins. The use of ingredients in the field is poor.
이러한 배경 아래에서, 본 발명자들은 카테킨 성분의 유용성에 대해 그 효용을 더욱 증대시키기 위해 예의 연구 노력한 결과, 카테킨에 현미, 서목태, 및 효모의 3종 복합추출물을 배합하는 경우, 카테킨의 항산화 효과가 더욱 개선되고, α-아밀라아제 효소 및 췌장 지방분해효소의 저해 활성이 더욱 증가하며, 지방전구세포(preadipocyte)의 분화 억제 효과가 더욱 증가하여 항비만 활성에 뚜렷하게 개선되는 시너지 효과가 있음을 확인하고 본 발명을 완성하게 되었다.Under this background, the present inventors have made intensive research efforts to further increase the utility of the catechin component, and as a result of mixing catechin with three complex extracts of brown rice, seomoktae, and yeast, the antioxidant effect of catechin is further enhanced. Improved, the inhibitory activity of α-amylase enzyme and pancreatic lipolytic enzyme is further increased, and the effect of inhibiting differentiation of predipocytes is further increased, thereby confirming that there is a synergistic effect of remarkably improving anti-obesity activity, and the present invention Was completed.
따라서, 본 발명의 주된 목적은 카테킨과 현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는 항비만용 조성물을 제공하는 데 있다.Accordingly, the main object of the present invention is to provide an anti-obesity composition comprising a complex extract of catechin and brown rice, seomoktae, and yeast as an active ingredient.
본 발명의 다른 목적 및 이점은 하기의 발명의 상세한 설명, 청구범위 및 도면에 의해 보다 명확하게 된다.Other objects and advantages of the present invention will become more apparent by the following detailed description, claims and drawings.
본 발명의 한 양태에 따르면, 본 발명은 카테킨과 현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는 항비만용 조성물을 제공한다.According to one aspect of the present invention, the present invention provides an anti-obesity composition comprising a complex extract of catechin and brown rice, seomoktae, and yeast as an active ingredient.
본 발명자들은 자연계에서 분리된 카테킨 성분의 유용성에 관하여 더욱 그 효용을 더욱 증대시키기 위해 연구를 수행한 결과, 카테킨에 현미, 서목태, 및 효모의 3종 복합추출물을 배합하는 경우, 종래 카테킨의 항산화 효과가 더욱 개선되고, α-아밀라아제 효소 및 췌장 지방분해효소의 저해 활성이 더욱 증가하며, 지방전구세포(preadipocyte)의 분화 억제 효과가 더욱 증가하게 되어 항비만 활성에 뚜렷하게 개선되는 시너지 효과가 있음을 확인하고 본 발명을 완성하게 되었다.The present inventors conducted a study to further increase the utility of the catechin component isolated from the natural world. As a result, when the three complex extracts of brown rice, Seomoktae, and yeast are mixed with catechin, the antioxidant effect of conventional catechins Is further improved, the inhibitory activity of α-amylase enzyme and pancreatic lipolytic enzyme is further increased, and the effect of inhibiting differentiation of predipocytes is further increased, confirming that there is a synergistic effect of remarkably improving anti-obesity activity. And completed the present invention.
본 발명에서 용어 "카테킨(Catechin)"은 하기 [화학식 1]로 표시되는 화합물로, 화학식은 [C15H14O6] 이고, 분자량은 290.27g/mol의 화합물이다. In the present invention, the term "catechin (Catechin)" is a compound represented by the following [Formula 1], the formula is [C 15 H 14 O 6 ], the molecular weight is a compound of 290.27g/mol.
[화학식 1] [Formula 1]
또한, 본 발명에서 용어 "현미"는 수확한 벼를 건조, 탈곡한 후 고무 롤러로 된 기계로 왕겨를 벗긴 쌀을 의미하며, 바람직하게는 5분 도미 내지 9분 도미를 사용할 수 있다. 또한, 용어 "서목태(Rhynchosia Nulubilis)"는 콩과의 다년생 만초의 열매로, "쥐눈이콩" 또는 "여두"로도 호칭된다. 또한, 효모(Saccharomyces cerevisiae)는 균(fungus) 계에 속하는 단세포의 진핵 미생물을 의미한다. In addition, in the present invention, the term "brown rice" refers to rice obtained by drying and threshing the harvested rice, and then peeling the rice husk by a machine with a rubber roller, and preferably, 5 minutes to 9 minutes sea bream may be used. In addition, the term " Rhynchosia Nulubilis" is a fruit of a perennial mancho in the legume family, and is also referred to as a "mouse-eyed bean" or "pollus". In addition, yeast ( Saccharomyces cerevisiae ) refers to a single-celled eukaryotic microorganism belonging to the fungus family.
본 발명에서는 상기 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물을 혼합하여 사용하는데, 용어 "카테킨 복합추출물"은 카테킨과 3종 복합추출물의 혼합물과 동일한 의미로 사용된다.In the present invention, the catechin and brown rice, seomoktae, and three kinds of complex extracts of yeast are mixed and used, and the term "catechin complex extract" is used in the same meaning as a mixture of catechin and three kinds of complex extracts.
본 발명에서 사용되는 용어, "추출물"은 원료의 추출 처리에 의하여 얻어지는 추출액, 상기 추출액의 희석액이나 농축액, 상기 추출액을 건조하여 얻어지는 건조물, 상기 추출액의 조정제물이나 정제물, 또는 이들의 혼합물 등, 추출액 자체 및 추출액을 이용하여 형성 가능한 모든 제형의 추출물을 포함한다. 본 발명의 상기 추출물은 바람직하게 추출 후 건조 분말 형태로 제조되어 사용될 수 있다.The term "extract" as used in the present invention refers to an extract obtained by extracting raw materials, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a preparation or purified product of the extract, or a mixture thereof, It includes the extract itself and extracts of all formulations that can be formed using the extract. The extract of the present invention may be preferably prepared and used in the form of a dry powder after extraction.
또한, 본 명세서에서 "유효성분"이란 단독으로 목적하는 활성을 나타내거나, 또는 그 자체는 활성이 없는 담체와 함께 활성을 나타낼 수 있는 성분을 의미한다.In addition, in the present specification, the term "active ingredient" refers to an ingredient that exhibits a desired activity by itself, or that itself can exhibit activity together with an inactive carrier.
본 발명의 용어 "예방"은 본 발명에 따른 조성물의 투여로 상기 비만 또는 비만 관련 질환의 발병을 억제 또는 지연시키는 모든 행위를 의미하고, 용어 "치료"는 본 발명에 따른 조성물의 투여로 상기 비만 또는 비만 관련 질환의 증세가 호전되거나 이롭게 변경하는 모든 행위를 의미한다.The term "prevention" of the present invention refers to any action that suppresses or delays the onset of obesity or obesity-related diseases by administration of the composition according to the present invention, and the term "treatment" refers to the obesity by administration of the composition according to the present invention. Or it refers to any action that improves or beneficially alters the symptoms of obesity-related diseases.
본 발명의 항비만용 조성물에서, 상기 카테킨과 상기 3종 복합추출물은 이에 제한되는 것은 아니나 바람직하게는 6 : 0.5 내지 1.5의 중량비로 배합될 수 있고, 더욱 바람직하게는 6:1의 중량비로 배합될 수 있다. 본 발명의 바람직한 실시예에서는 상기 카테킨과, 상기 3종 복합추출물이 6:1의 중량비로 배합된 혼합물에서 우수한 항산화 효과가 있고, α-아밀라아제 효소 및 췌장 지방분해효소의 저해 활성이 증가하며, 지방전구세포(preadipocyte)의 분화 억제효과가 증가하여 항비만 활성에 뚜렷하게 개선되는 시너지 효과가 있음을 확인한 바 있다. 또한, 상기 현미 추출물, 서목태 추출물, 및 효모 추출물들은 바람직하게 가장 많이 투입된 어느 하나의 성분 100 중량부 대비 다른 성분의 혼합되는 중량부가 30 이상 100 미만으로 혼합되는 것이 전체 복합추출물이 카테킨과 시너지 효과를 나타낼 수 있어 바람직하며, 가장 바람직하게는 이들 3종 추출물들을 동량으로 혼합하는 것이 카테킨과 시너지 효과를 내는 측면에서 가장 바람직하다.In the composition for anti-obesity of the present invention, the catechin and the three complex extracts are not limited thereto, but are preferably blended in a weight ratio of 6: 0.5 to 1.5, and more preferably blended in a weight ratio of 6: 1. Can be. In a preferred embodiment of the present invention, a mixture of the catechin and the three complex extracts in a weight ratio of 6:1 has an excellent antioxidant effect, increases the inhibitory activity of α-amylase enzyme and pancreatic lipolytic enzyme, and increases fat It has been confirmed that there is a synergistic effect of remarkably improving antiobesity activity by increasing the differentiation inhibitory effect of progenitor cells (preadipocyte). In addition, the brown rice extract, seomoktae extract, and yeast extracts are preferably mixed in an amount of 30 to less than 100 parts by weight of the other ingredients compared to 100 parts by weight of any one component added the most, so that the entire complex extract has a synergistic effect with catechins. It is preferable because it can be represented, and most preferably, mixing these three extracts in the same amount is most preferable in terms of producing a synergistic effect with catechin.
본 발명의 항비만용 조성물은 항산화 활성을 갖는 것을 특징으로 한다. 본 발명의 바람직한 실시예에서는 본 발명의 카테킨 복합추출물이 처리된 실험군은 3종 복합추출물의 첨가 없이 카테킨을 단독으로 처리한 대조군에 비하여 전반적으로 우수한 항산화 효과를 보이는 것을 관찰할 수 있었다(도 1 및 도 2). 이러한 항산화 효과 수치들은 양성대조군인 아스코르브산 처리군과 비교하였을 때도 비슷한 수준인 것으로 분석되었다. 또한, 카테킨 복합추출물은 지방세포 내에서도 산화 스트레스에 대해 뛰어난 세포보호 효과를 나타내는 것을 확인할 수 있었다(도 3).The composition for anti-obesity of the present invention is characterized by having antioxidant activity. In a preferred embodiment of the present invention, it was observed that the experimental group treated with the catechin complex extract of the present invention exhibited an overall superior antioxidant effect compared to the control group treated with catechin alone without the addition of the three complex extracts (Fig. 1 and Fig. 2). These antioxidant effect levels were analyzed to be similar when compared to the positive control group, ascorbic acid treatment group. In addition, it was confirmed that the catechin complex extract exhibited an excellent cytoprotective effect against oxidative stress even in adipocytes (FIG. 3).
또한, 본 발명의 항비만용 조성물은 α-아밀라아제 효소 및 췌장 지방분해효소의 활성을 저해하는 것을 특징으로 한다. 본 발명의 바람직한 실시예에서는 본 발명의 카테킨 복합추출물은 500ug/ml 농도에서 α-아밀라아제 활성을 80% 이상 억제하는 것을 확인할 수 있었고(도 4), 카테킨을 단독처리군에 비해 모든 구간에서 췌장 리파아제 효소의 저해 활성이 증가하여 카테킨과 3종 복합추출물 사이의 시너지 효과가 있음을 확인하였다(도 5). In addition, the composition for anti-obesity of the present invention is characterized by inhibiting the activity of α-amylase enzyme and pancreatic lipolytic enzyme. In a preferred embodiment of the present invention, it was confirmed that the catechin complex extract of the present invention inhibited α-amylase activity by 80% or more at a concentration of 500 ug/ml (FIG. 4), and pancreatic lipase in all sections compared to the group treated with catechin alone. It was confirmed that there was a synergistic effect between the catechin and the three complex extracts by increasing the inhibitory activity of the enzyme (FIG. 5).
본 발명의 항비만용 조성물은 지방전구세포(preadipocyte)의 분화 억제를 통해 항비만 활성이 있는 갖는 것을 특징으로 한다. 본 발명의 바람직한 실시예에서는 카테킨 500ug/ml 단일 처리군에서 26% 억제 활성을 보인 것에 비해, 본 발명의 카테킨 복합추출물 500ug/ml 처리군에서는 34% 수분의 지방세포 분화 억제율이 관찰되어 카테킨 단일처리군에 비해 31% 개선된 지방세포 분화 억제 효과를 확인하였다(도 7).The composition for anti-obesity of the present invention is characterized by having anti-obesity activity through inhibition of differentiation of predipocytes. In a preferred embodiment of the present invention, the
본 발명의 일 양태에 따르면, 본 발명은 카테킨과, 현미, 서목태, 및 효모의 3종 복합추출물을 유효성분으로 포함하는 항비만용 약학적 조성물을 제공한다. 본 발명의 약학적 조성물은 우수한 항산화 효과가 있고, α-아밀라아제 효소 및 췌장 지방분해효소의 저해 활성이 우수하며, 지방전구세포(preadipocyte)의 분화 억제에 우수한 효과가 있으므로, 비만의 치료 및 예방과 관련한 항비만용 약학적 조성물로 유용하게 사용될 수 있다.According to one aspect of the present invention, the present invention provides a pharmaceutical composition for anti-obesity comprising three types of complex extracts of catechin, brown rice, seomoktae, and yeast as an active ingredient. The pharmaceutical composition of the present invention has an excellent antioxidant effect, has excellent inhibitory activity of α-amylase enzyme and pancreatic lipolytic enzyme, and has excellent effect on inhibiting differentiation of predipocytes, thus treating and preventing obesity and It can be usefully used as a related anti-obesity pharmaceutical composition.
한편, 본 발명의 카테킨과 현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는 항비만용 약학적 조성물은 약학적으로 허용 가능한 염의 형태로 제조될 수 있다. Meanwhile, the pharmaceutical composition for anti-obesity comprising the complex extract of catechin and brown rice, Seomoktae, and yeast of the present invention as an active ingredient may be prepared in the form of a pharmaceutically acceptable salt.
구체적으로 산을 첨가함으로써 염을 형성할 수 있고, 예를 들어 무기산(예: 염산, 히드로브롬산, 인산, 질산, 황산 등), 유기 카르복실산(예: 아세트산, 트리플루오로아세트산과 같은 할로 아세트산, 프로피온산, 말레산, 숙신산, 말산, 시트르산, 타르타르산, 살리실산), 및 산성 당(글루쿠론산, 갈락투론산, 글루콘산, 아스코르브산), 산성 폴리사카리드(예: 히알우론산, 콘드로이틴 술페이트, 아르기닌산), 콘드로이틴 술페이트와 같은 술폰산 당 에스테르를 포함하는 유기 술폰산(예: 메탄, 술폰산, p-톨루엔 술폰산) 등을 첨가하여 염을 형성할 수 있다.Specifically, salts can be formed by adding an acid, for example, inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, etc.), organic carboxylic acids (such as acetic acid and trifluoroacetic acid). Acetic acid, propionic acid, maleic acid, succinic acid, malic acid, citric acid, tartaric acid, salicylic acid), and acidic sugars (glucuronic acid, galacturonic acid, gluconic acid, ascorbic acid), acidic polysaccharides (e.g. hyaluronic acid, chondroitin sulfate, Arginic acid), an organic sulfonic acid (eg, methane, sulfonic acid, p-toluene sulfonic acid) including a sulfonic acid sugar ester such as chondroitin sulfate may be added to form a salt.
본 발명의 약학적 조성물은 세포독성이 없는 천연 추출물에서 분리되는 물질들로 이루어져 임상투여시 경구 또는 비경구로 투여가 가능하며, 일반적인 의약품 제제의 형태로 사용될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally during clinical administration, consisting of substances isolated from natural extracts without cytotoxicity, and may be used in the form of general pharmaceutical preparations.
즉, 본 발명의 약학적 조성물은 실제로 경구 또는 비경구의 여러 가지 제형으로 투여될 수 있는데, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 비경구투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 리우린지, 글리세로제라틴 등이 사용될 수 있다.That is, the pharmaceutical composition of the present invention can actually be administered in various oral or parenteral formulations, but when formulated, diluents or excipients such as commonly used fillers, extenders, binders, wetting agents, disintegrants, and surfactants are used. It is prepared by doing. Preparations for parenteral administration include sterilized aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for the suppository, witepsol, macrogol, tween 61, cacao butter, liurinji, glycerogelatin, and the like may be used.
또한, 본 발명의 약학적 조성물은 생리식염수 또는 유기용매와 같이 약제로 허용된 여러 전달체(carrier)와 혼합하여 사용될 수 있고, 안정성이나 흡수성을 증가시키기 위하여 글루코스, 수크로스 또는 덱스트란과 같은 카보하이드레이트, 아스코르브 산(ascorbic acid) 또는 글루타치온과 같은 항산화제(antioxidants), 킬레이트화제(chelating agents), 저분자 단백질 또는 다른 안정화제(stabilizers)들이 약제로 사용될 수 있다.In addition, the pharmaceutical composition of the present invention may be used by mixing with various carriers permitted as drugs such as physiological saline or an organic solvent, and carbohydrates such as glucose, sucrose or dextran to increase stability or absorption. , Antioxidants such as ascorbic acid or glutathione, chelating agents, small molecule proteins or other stabilizers may be used as pharmaceuticals.
본 발명의 약학적 조성물의 유효용량은 0.01㎎/㎏ 내지 10㎎/㎏이고, 바람직하게는 0.1㎎/㎏ 내지 1㎎/㎏ 이며, 하루 1회 내지 3회 투여될 수 있다.The effective dose of the pharmaceutical composition of the present invention is 0.01 mg/kg to 10 mg/kg, preferably 0.1 mg/kg to 1 mg/kg, and may be administered once to three times a day.
본 발명의 약학적 조성물의 총 유효량은 볼루스(bolus) 형태 혹은 상대적으로 짧은 기간 동안 주입(infusion) 등에 의해 단일 투여량(single does)으로 환자에게 투여될 수 있으며, 다중 투여량(multiple does)이 장기간 투여되는 분할 치료 방법(fractionated treatment protocol)에 의해 투여될 수 있다. 상기 농도는 약의 투여 경로 및 치료 횟수뿐만 아니라 환자의 나이 및 건강상태 등 다양한 요인들을 고려하여 환자의 유효 투여량이 결정되는 것이므로 이러한 점을 고려할 때, 이 분야의 통상적인 지식을 가진 자라면 본 발명의 약학적 조성물로서의 특정한 용도에 따른 적절한 유효 투여량을 결정할 수 있을 것이다.The total effective amount of the pharmaceutical composition of the present invention can be administered to a patient in the form of a bolus or in a relatively short period of time by infusion or the like as a single dose, and multiple doses It can be administered by this long-term, fractionated treatment protocol. Since the concentration is determined by taking into account various factors such as the patient's age and health status, as well as the route of administration and the number of treatments of the drug, the effective dosage of the patient is determined. It will be possible to determine an appropriate effective dosage according to the specific use of the pharmaceutical composition.
본 발명의 다른 양태에 따르면, 본 발명은 카테킨과 현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는 항비만용 식품 조성물을 제공한다. 본 발명의 카테킨과, 현미 추출물, 서목태 추출물, 및 효모 추출물은 상기에서 설명한 바와 같다. 상기 식품 조성물은 건강기능식품의 형태로 사용될 수 있으나, 이에 제한되는 것은 아니다.According to another aspect of the present invention, the present invention provides an anti-obesity food composition comprising a complex extract of catechin and brown rice, seomoktae, and yeast as an active ingredient. The catechin, brown rice extract, seomoktae extract, and yeast extract of the present invention are as described above. The food composition may be used in the form of a health functional food, but is not limited thereto.
본 발명의 식품 조성물은 카테킨과 현미, 서목태, 및 효모의 복합추출물, 이의 분획물 또는 이의 가공물의 형태로 포함될 수 있다. 또한, 상기 조성물은 유효성분 이외에 식품학적으로 허용 가능한 식품보조첨가제를 포함할 수 있다.The food composition of the present invention may be included in the form of a complex extract of catechin and brown rice, seomoktae, and yeast, a fraction thereof, or a processed product thereof. In addition, the composition may contain food additives that can be food physiologically acceptable in addition to the active ingredient.
본 발명에 있어서, "식품보조첨가제"란 식품에 보조적으로 첨가될 수 있는 구성요소를 의미하며, 각 제형의 건강기능식품을 제조하는데 첨가되는 것으로서 당업자가 적절히 선택하여 사용할 수 있다. 식품보조첨가제의 예로는 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 충진제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등이 포함되지만, 상기 예들에 의해 본 발명의 식품보조첨가제의 종류가 제한되는 것은 아니다.In the present invention, the term "food supplementary additive" refers to a component that can be added auxiliary to food, and is added to the manufacture of health functional foods of each formulation, and can be appropriately selected and used by those skilled in the art. Examples of food additives include various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavors and natural flavoring agents, coloring agents and fillers, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners. , pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, etc. are included, but the types of food additives of the present invention are not limited by the above examples.
본 발명의 식품 조성물에는 건강기능식품이 포함될 수 있다. 본 발명에 있어서, "건강기능식품"이란 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 정제, 캅셀, 분말, 과립, 액상 및 환 등의 형태로 제조 및 가공한 식품을 말한다. 여기서 “기능성”이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻는 것을 의미한다. 본 발명의 건강기능식품은 당업계에서 통상적으로 사용되는 방법으로 제조 가능하며, 상기 제조시에는 당업계에서 통상적으로 첨가하는 원료 및 성분을 첨가하여 제조할 수 있다. The food composition of the present invention may contain a health functional food. In the present invention, "health functional food" refers to food manufactured and processed in the form of tablets, capsules, powders, granules, liquids and pills using raw materials or ingredients having useful functions for the human body. Here, "functionality" means obtaining useful effects for health purposes such as controlling nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be prepared by a method commonly used in the art, and during the production, raw materials and ingredients commonly added in the art may be added to prepare it.
또한, 상기 건강기능식품의 제형 또한 건강기능식품으로 인정되는 제형이면 제한 없이 제조될 수 있다. 본 발명의 식품 조성물은 다양한 형태의 제형으로 제조될 수 있으며, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용 시 발생할 수 있는 부작용 등이 없는 장점이 있고, 휴대성이 뛰어나, 비만을 개선하는 효과를 증진시키기 위한 보조제로 섭취가 가능하다.In addition, the formulation of the health functional food may be prepared without limitation as long as it is a formulation recognized as a health functional food. The food composition of the present invention can be prepared in a variety of forms, and unlike general drugs, it has the advantage of not having side effects that may occur when taking the drug for a long time using food as a raw material, and is excellent in portability and improves obesity. It can be taken as an adjuvant to enhance the effect.
본 발명의 건강기능식품이 취할 수 있는 형태에는 제한이 없으며, 통상적인 의미의 식품을 모두 포함할 수 있고, 기능성 식품 등 당업계에 알려진 용어와 혼용이 가능하다. 아울러 본 발명의 건강기능식품은 당업자의 선택에 따라 식품에 포함될 수 있는 적절한 기타 보조 성분과 공지의 첨가제를 혼합하여 제조할 수 있다. 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 등이 있으며, 본 발명에 따른 화학식 1로 표시되는 화합물을 주성분으로 하여 제조한 즙, 차, 젤리 및 주스 등에 첨가하여 제조할 수 있다. 또한, 동물을 위한 사료로 이용되는 식품도 포함된다.There is no limitation on the form that the health functional food of the present invention can take, and may include all foods in a conventional sense, and may be mixed with terms known in the art such as functional foods. In addition, the health functional food of the present invention can be prepared by mixing appropriate other auxiliary ingredients and known additives that may be included in the food according to the choice of a person skilled in the art. Examples of foods that can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and There are vitamin complexes and the like, and can be prepared by adding to juice, tea, jelly, juice, etc. prepared using the compound represented by
본 발명의 특징 및 이점을 요약하면 다음과 같다:The features and advantages of the present invention are summarized as follows:
(a) 본 발명은 카테킨과 현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는 항비만용 조성물을 제공한다.(a) The present invention provides an anti-obesity composition comprising a complex extract of catechin and brown rice, seomoktae, and yeast as an active ingredient.
(b) 본 발명의 항비만용 조성물은 카테킨 단일 성분에 비해 개선된 항산화 효과를 나타내고, α-아밀라아제 효소 및 췌장 지방분해효소의 저해 활성이 우수하며, 지방전구세포(preadipocyte)의 분화 억제에 우수한 효과가 있다.(b) The composition for anti-obesity of the present invention exhibits improved antioxidant effect compared to a single component of catechin, has excellent inhibitory activity of α-amylase enzyme and pancreatic lipolytic enzyme, and is excellent in inhibiting differentiation of predipocytes. It works.
(c) 또한, 본 발명의 항비만용 조성물은 천연에서 존재하는 식용물질인 카테킨과 현미, 서목태, 및 효모 추출물들을 배합하여 인체에 매우 안전할 뿐만 아니라, 안정성도 매우 탁월하여 식품, 및 의약품 분야에서 비만의 치료 및 예방과 관련한 항비만용 목적으로 유용하게 이용 가능하다. (c) In addition, the composition for anti-obesity of the present invention is not only very safe for the human body, but also very excellent in stability by combining natural edible substances such as catechin and brown rice, seomoktae, and yeast extract. It can be usefully used for anti-obesity purposes related to the treatment and prevention of obesity.
도 1은 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 DPPH 라디칼 소거 활성을 측정하여 나타낸 그래프이다.
도 2는 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 ABTS 라디칼 소거 활성을 측정하여 나타낸 그래프이다.
도 3은 지방전구세포인 3T3-L1 세포에서 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 세포 내 ROS 소거 활성을 측정하여 나타낸 그래프이다.
도 4는 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 α-아밀라아제 효소 활성 억제율을 측정하여 나타낸 그래프이다.
도 5는 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 췌장 지방분해효소 활성 억제율을 측정하여 나타낸 그래프이다.
도 6은 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 지방전구세포인 3T3-L1 세포의 세포 생존율을 측정하여 나타낸 그래프이다.
도 7은 지방전구세포인 3T3-L1 세포에서 본 발명의 카테킨과 현미, 서목태, 및 효모의 3종 복합추출물(복합물)의 처리 농도에 따른 지방전구세포(preadipocyte)의 분화 억제율을 측정하여 나타낸 그래프이다.1 is a graph showing DPPH radical scavenging activity according to the treatment concentration of three complex extracts (complexes) of catechin and brown rice, Seomoktae, and yeast of the present invention.
Figure 2 is a graph showing the ABTS radical scavenging activity according to the treatment concentration of the catechin and brown rice, Seomoktae, and the three complex extracts (composite) of yeast of the present invention.
3 is a graph showing the measurement of intracellular ROS scavenging activity according to the treatment concentration of the catechin of the present invention and the three complex extracts (complexes) of brown rice, Seomoktae, and yeast in 3T3-L1 cells, which are adipocytes.
4 is a graph showing the inhibition rate of α-amylase enzyme activity according to the treatment concentration of the three complex extracts (complexes) of catechin and brown rice, Seomoktae, and yeast of the present invention.
Figure 5 is a graph showing the measurement of the inhibition rate of pancreatic lipolytic enzyme activity according to the treatment concentration of three complex extracts (complexes) of catechin and brown rice, Seomoktae, and yeast of the present invention.
6 is a graph showing the cell viability of 3T3-L1 cells, which are adipocytes, according to the treatment concentration of the three complex extracts (complexes) of catechin and brown rice, Seomoktae, and yeast according to the present invention.
Figure 7 is a graph showing the differentiation inhibition rate of fat progenitor cells (preadipocyte) according to the treatment concentration of the catechin of the present invention and the three kinds of complex extracts (complexes) of brown rice, Seomoktae, and yeast in 3T3-L1 cells, which are adipocytes. to be.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하기로 한다. 이들 실시예는 단지 본 발명을 예시하기 위한 것이므로, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는다.Hereinafter, the present invention will be described in more detail through examples. Since these examples are for illustrative purposes only, the scope of the present invention is not to be construed as being limited by these examples.
실시예 1. 카테킨 복합추출물의 제조 Example 1. Preparation of catechin complex extract
카테킨과 효모는 동서바이오팜(용인, 경기도)에서 구입하여 사용하였다. Catechin and yeast were purchased and used from Dongseo Biopharm (Yongin, Gyeonggi-do).
또한, 5분도 현미 쌀 1kg을 분쇄한 후 20L의 70% 에탄올을 용매로 사용하여 12시간 가온 환류추출한 후, 냉침 시키고, 1.2㎛ 투과 사이즈를 갖는 여과지로 여과하여 여과액을 수득하였다. 수득된 여과액을 감압농축 및 감압 건조로 농축하여 쌀 건조 분말을 제조하였다.In addition, 1 kg of brown rice was pulverized for 5 minutes, heated to reflux extraction for 12 hours using 20 L of 70% ethanol as a solvent, cooled, and filtered through a filter paper having a transmission size of 1.2 μm to obtain a filtrate. The obtained filtrate was concentrated under reduced pressure and dried under reduced pressure to prepare dry rice powder.
상기 현미 건조 분말을 제조하는 방법과 동일한 방법을 사용하여 서목태와 효모 건조 분말을 제조하였다. 이렇게 제조된 현미, 서목태, 및 효모의 3종 건조 분말을 동량으로 혼합하여 복합추출물을 제조하였다. Seomoktae and yeast dry powder were prepared using the same method as the method of preparing the dried brown rice powder. A complex extract was prepared by mixing the thus prepared brown rice, Seomoktae, and three dry powders of yeast in equal amounts.
이렇게 준비된 카테틴과, 현미 추출물, 서목태 추출물, 및 효모의 3종 복합추출물을 6:1의 중량비로 혼합하여 최종 카테킨 복합추출물을 제조하였다. The prepared catetin, brown rice extract, seomoktae extract, and three complex extracts of yeast were mixed at a weight ratio of 6:1 to prepare a final catechin complex extract.
실시예 2. 카테킨 복합추출물의 항산화 활성Example 2. Antioxidant activity of catechin complex extract
2-1. DPPH 라디칼 소거 활성2-1. DPPH radical scavenging activity
산화적 스트레스는 신경세포의 신호전달 체계를 작동시키고 과산화수소 등의 활성산소종의 발생을 증가시키며, 항산화 효소의 발현을 감소시킨다. 이러한 활성산소종의 증가는 유전자와 단백질의 구조를 변화시켜 세포 내 및 세포 외의 항상성이 파괴되어 신경세포의 손상을 유발한다. 이와 같은 신경세포의 손상을 유발하는 활성산소종의 하나인 DPPH radical의 억제 활성은 항산화 효능을 평가하는 중요한 지표가 된다.Oxidative stress activates the signaling system of nerve cells, increases the generation of reactive oxygen species such as hydrogen peroxide, and decreases the expression of antioxidant enzymes. This increase in reactive oxygen species changes the structure of genes and proteins, thereby destroying intracellular and extracellular homeostasis, causing damage to nerve cells. The inhibitory activity of DPPH radical, one of the reactive oxygen species that induces nerve cell damage, is an important index for evaluating antioxidant efficacy.
DPPH(1,1-diphenyl-2-picrylhydrazyl, Sigma, USA)의 환원력을 이용하는 상등의 문헌(Sang S et al., J.Agric. Food Chem. 50(8), p2459-2463, 2002)에 기재된 방법에 따라, 본 발명에 따른 카테킨 복합추출물의 항산화 활성을 측정하였다.The superior literature using the reducing power of DPPH (1,1-diphenyl-2-picrylhydrazyl, Sigma, USA) (Sang S et al., J. Agric. Food Chem. 50(8), p2459-2463, 2002). According to the method, the antioxidant activity of the catechin complex extract according to the present invention was measured.
실시예 1에서 준비한 카테킨 복합추출물(카테킨: 3종 복합추출물= 6:1 중량비)을 각각 1, 10, 50, 100, 및 250㎍/㎖ 농도로 96-웰마이크로 플레이트에 넣고 37℃에서 30분간 배양한 후, 환원된 DPPH의 양을 분광 광도계(Precision microplate reader; Molecular Devices, VersaMax)를 이용하여 520nm에서 흡광도를 측정하여 시료를 가하지 않는 무첨가 대조군과 활성을 비교하였다. 상기 시험은 3회 반복하였으며, 그 결과를 도 1에 나타내었다. 현미, 서목태, 및 효모의 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군으로 사용하였고, 비교 실험을 위해 아스코르브산(Ascorbic acid)을 양성대조군으로 사용하였다.The catechin complex extract prepared in Example 1 (catechin: 3 types of complex extract = 6:1 weight ratio) was placed in a 96-well microplate at concentrations of 1, 10, 50, 100, and 250 μg/ml, respectively, at 37° C. for 30 minutes. After incubation, the amount of reduced DPPH was measured by measuring the absorbance at 520 nm using a spectrophotometer (Precision microplate reader; Molecular Devices, VersaMax), and the activity was compared with an additive-free control group without adding a sample. The test was repeated three times, and the results are shown in FIG. 1. The group treated with catechin alone without the three complex extracts of brown rice, Seomoktae, and yeast was used as a control group, and ascorbic acid was used as a positive control group for the comparative experiment.
그 결과, 본 발명의 카테킨 복합추출물이 처리된 실험군은 3종 복합추출물의 첨가 없이 카테킨을 단독으로 처리한 대조군에 비하여 전반적으로 매우 우수한 항산화 효과를 보이는 것을 관찰할 수 있었고, 이러한 항산화 효과 수치들은 양성대조군인 아스코르브산 처리군과 비교하였을 때도 비슷한 수준인 것으로 나타났으며, 매우 우수한 항산화 활성이 있는 것으로 분석되었다. As a result, it was observed that the experimental group treated with the catechin complex extract of the present invention exhibited a very superior antioxidant effect overall compared to the control group treated with catechin alone without the addition of the three complex extracts, and these antioxidant effect values were positive. Compared to the control group treated with ascorbic acid, it was found to be at a similar level, and it was analyzed that it has very good antioxidant activity.
2-2. ABTS 라디칼 소거 활성2-2. ABTS radical scavenging activity
2,2'-아지노-비스(3-에틸벤조티아졸린-6-술폰산)(ABTS) 라디칼 양이온 소거능은 디암모늄염과 과황화칼륨과의 반응으로 ABTS 라디칼이 생성되면 특유의 색인 청록색을 띄게 되는데, 항산화 효과를 나타내는 시료를 첨가함에 따라 연한 녹색으로 탈색(decolorization)되는 것을 측정하는 방법이다.The 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation scavenging ability becomes a distinctive blue-green color when the ABTS radical is generated by the reaction of a diammonium salt and potassium persulfide. This is a method of measuring decolorization to light green as a sample showing an antioxidant effect is added.
ABTS 라디칼을 이용한 항산화능 측정 시험은 7mM 2,2'-아지노-bis(3-에틸벤조티아졸린-6-술폰산)과 2.4 mM 과황화칼륨을 혼합하여 사용 전 16시간 동안 빛을 차단하고 실온에서 반응시켰다. 반응 후 ABTS 1ml(in PBS, pH 7.4)에 상기 실시예 1에 따라 준비된 카테킨 복합추출물(카테킨: 3종 복합추출물= 6:1 중량비)을 각각 1, 10, 50, 100, 및 250㎍/㎖(in PBS, pH 7.4) 농도로 첨가하여 반응시킨 후, 10분 뒤 원심분리하고, 상층액만 취득하여 734nm에서 흡광도를 측정하였다. 현미, 서목태, 및 효모의 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군으로 사용하였고, 비교 실험을 위해 아스코르브산(Ascorbic acid)을 양성대조군으로 사용하였다.Antioxidant activity measurement test using ABTS radical was conducted by mixing
측정 결과, 본 발명의 카테킨 복합추출물이 처리된 실험군은 식물 복합추출물 없이 카테킨을 단독으로 처리한 대조군에 비하여 모든 구간에서 매우 우수한 항산화 효과를 보이는 것을 관찰할 수 있었고, 이러한 항산화 효과 수치들은 양성대조군인 아스코르브산 처리군과 비교하였을 때도 동등 이상의 수준인 것으로 나타났으며, 매우 우수한 항산화 활성을 갖는 것으로 분석되었다. As a result of the measurement, it was observed that the experimental group treated with the catechin complex extract of the present invention showed very excellent antioxidant effect in all sections compared to the control group treated with catechin alone without the plant complex extract. When compared with the ascorbic acid treatment group, it was found to be equal or higher than that of the group, and it was analyzed to have very excellent antioxidant activity.
특히, 50㎍/㎖ 농도 이상 구간에서 양성대조군인 아스코르브산 처리군 또는 카테킨 단독 처리군 보다 월등하게 우수한 항산화 효과를 나타내는 점은 주목할만한 특징으로 보이며, 이러한 강력한 항산화 활성은 본 발명의 카테킨 복합추출물을 소재로 이용하여 상품화할 때 유용할 것으로 사료된다. In particular, it seems to be a remarkable feature that it exhibits a far superior antioxidant effect than the positive control group treated with ascorbic acid or the group treated with catechin alone at a concentration of 50 µg/ml or higher, and such strong antioxidant activity makes the catechin complex extract of the present invention. It is believed to be useful when commercialized by using it as a material.
2-3. 세포 내 라디칼 소거 활성2-3. Intracellular radical scavenging activity
DCF-DA(2',7'-dichlorodihydrofluorescein diacetate)를 이용하여 세포 내 활성산소종 수준을 측정하였다. DCF-DA는 활성산소종에 의해 세포 내에서 DCF로 전환되어 형광을 발한다. Intracellular reactive oxygen species levels were measured using DCF-DA (2',7'-dichlorodihydrofluorescein diacetate). DCF-DA is converted to DCF in cells by reactive oxygen species and fluoresces.
3T3-L1 지방전구세포(preadipocyte)(ATCC CL-173, American Type Culure Collection, Manassas, USA)를 5% FBS(fetal bovine serum), 10% horse serum, 50units/mL penicillin 및 100μg/mL streptomycin을 포함하는 DMEM 배지에 접종하여 37℃, 5% CO2 조건에서 24∼48시간 계대 배양하여 실험에 사용하였다.3T3-L1 fat progenitor cells (ATCC CL-173, American Type Culure Collection, Manassas, USA) containing 5% FBS (fetal bovine serum), 10% horse serum, 50 units/mL penicillin and 100 μg/mL streptomycin It was inoculated in the DMEM medium and subcultured for 24 to 48 hours under conditions of 37°C and 5% CO 2, and used in the experiment.
3T3-L1 세포를 105 내지 106 농도로 분주하고, 본 발명의 카테킨 복합추출물(카테킨:3종 복합추출물= 6:1 중량비)을 7.81, 15.63, 31.25, 62.5, 125, 250, 및 500㎍/㎖ 농도로 처리하여 48시간 동안 배양하였다. 비교 실험을 위해 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군으로 사용하였다. 3T3-L1 cells were dispensed at a concentration of 10 5 to 10 6 , and the catechin complex extract (catechin: 3 types complex extract = 6:1 weight ratio) of the present invention was 7.81, 15.63, 31.25, 62.5, 125, 250, and 500 µg. It was treated at /ml concentration and incubated for 48 hours. For the comparative experiment, the group treated with catechin alone without the three complex extracts was used as a control.
배양 후, 0.5mM 농도의 H2O2를 4시간 동안 처리하였고, 다음 250μM DCF-DA를 넣어 50분 동안 배양하여 형광 마이크로 플레이트 리더(microplate reader, Molecular Devices E09090, USA)를 사용하여 485nm(determination)와 535nm(reference)에서 형광 레벨을 측정하였다.After incubation, 0.5 mM H 2 O 2 was treated for 4 hours, and then 250 μM DCF-DA was added and incubated for 50 minutes, and then 485 nm (determination) using a fluorescent microplate reader (Molecular Devices E09090, USA). ) And fluorescence levels were measured at 535nm (reference).
그 결과, 본 발명의 카테킨 복합추출물은 H2O2 처리로 생성된 3T3-L1 세포 내의 라디칼을 효과적으로 소거하는 것으로 나타났으며, 250ug/ml 이상의 농도에서 3T3-L1 세포 내 라디칼의 생성을 45% 이상 감소시키는 것으로 나타났다. 또한, 500ug/ml의 농도에서는 카테킨 단일 처리군에 비해 라디칼 소거 활성이 10% 이상 증가한 것으로 나타났다. 상기 결과를 통해 본 발명의 카테킨 복합추출물은 지방세포 내에서도 산화 스트레스에 대해 뛰어난 세포보호 효과를 나타내는 것을 확인할 수 있었다.As a result, the catechin complex extract of the present invention was found to effectively scavenge radicals in 3T3-L1 cells generated by H 2 O 2 treatment, and at a concentration of 250 ug/ml or more, the generation of radicals in 3T3-L1 cells was reduced by 45%. It has been shown to reduce abnormalities. In addition, at the concentration of 500ug/ml, the radical scavenging activity was increased by more than 10% compared to the catechin single treatment group. Through the above results, it was confirmed that the catechin complex extract of the present invention exhibits an excellent cytoprotective effect against oxidative stress even in adipocytes.
실시예 3. 카테킨 복합추출물의 α-아밀라아제 저해 활성Example 3. α-amylase inhibitory activity of catechin complex extract
당질 가수분해와 흡수 저해에 따른 식후 혈당 농도 조절 효과를 확인하기 위하여, 상기 실시예 1에서 제조한 카테킨 3종 복합추출물이 당가수분해 효소인 α-아밀라아제의 활성에 미치는 영향을 실험하였다. 구체적으로, 본 발명의 카테킨 복합추출물(카테킨:3종 복합추출물= 6:1 중량비)을 각각 62.5, 125, 250, 및 500㎍/㎖ 농도로 0.1M 인산나트륨 버퍼(sodium phosphate buffer)(pH 6.9) 및 0.5 unit/mL의 α-아밀라아제 효소 용액을 혼합하여 37℃에서 10분간 배양하였다. 배양 후, 200㎕ 5 mM [4-nitrophenyl-α-D-gluco-pyranoside] 화합물을 가하여 37℃에서 5분간 반응시킨 뒤 405nm에서 마이크로플레이트 리더기(microplate reader, Molecular Devices E09090, USA)로 흡광도를 측정하였다. 비교 실험을 위해 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군으로 사용하였다. In order to confirm the effect of adjusting postprandial blood sugar concentration by inhibition of sugar hydrolysis and absorption, the effect of the three catechin complex extract prepared in Example 1 on the activity of α-amylase, a sugar hydrolyzing enzyme, was tested. Specifically, the catechin complex extract of the present invention (catechin: 3 types of complex extract = 6:1 weight ratio) was 0.1M sodium phosphate buffer (pH 6.9) at a concentration of 62.5, 125, 250, and 500 μg/ml, respectively. ) And 0.5 unit/mL of α-amylase enzyme solution were mixed and incubated for 10 minutes at 37°C. After incubation, 200 μl 5 mM [4-nitrophenyl-α-D-gluco-pyranoside] compound was added and reacted at 37° C. for 5 minutes, and the absorbance was measured at 405 nm with a microplate reader (Molecular Devices E09090, USA). I did. For the comparative experiment, the group treated with catechin alone without the three complex extracts was used as a control.
그 결과, 본 발명의 카테킨 복합추출물은 500ug/ml 농도에서 α-아밀라아제 활성을 80% 이상 억제하는 것을 확인할 수 있었고, 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군에 비해 모든 구간에서 α-아밀라아제 저해 활성이 우수한 것을 확인하였다(도 4). 따라서, 상기 실험결과를 통해, 본 발명의 카테킨 복합추출물의 혈당 조절 소재로의 우수한 역할을 확인할 수 있었다.As a result, it was confirmed that the catechin complex extract of the present invention inhibited α-amylase activity by 80% or more at a concentration of 500 ug/ml, and the group treated with catechin alone without the three complex extracts was α in all sections compared to the control group. -It was confirmed that the amylase inhibitory activity was excellent (Fig. 4). Therefore, through the above experimental results, it was possible to confirm the excellent role of the catechin complex extract of the present invention as a blood sugar control material.
실시예 4. 카테킨 복합추출물의 췌장 지방분해효소 활성 저해Example 4. Inhibition of pancreatic lipolytic enzyme activity of catechin complex extract
지방은 췌장 리파아제에 의해 분해되어 흡수되기 때문에 리파아제 효소의 활성을 저해할 경우 혈중의 지방흡수를 저해시킬 수 있다. Since fat is decomposed and absorbed by pancreatic lipase, if the activity of the lipase enzyme is inhibited, it can inhibit the absorption of fat in the blood.
췌장 리파아제 저해활성 측정은 Kim 등이 행한 방법을 변형하여 실시하였다. 즉 효소 완충용액(10mM MOPS, 1mM EDTA, pH 6.8)에 돼지 췌장 리파아제(porcine pancreatic lipase)를 0.5g/200㎖의 농도로 4℃를 유지하면서 용해하고, 상기 용해물을 4000rpm으로 원심 분리한 후, 상층액에 트리스 완충용액(100mM Tris-HCl, 5mM CaCl2, pH 7.0)과 효소 완충용액(enzyme buffer)을 혼합하였다. 실시예 1에서 제조한 카테킨 복합추출물(카테킨: 3종 복합추출물= 6:1 중량비)을 각각 62.5, 125, 250, 및 500㎍/㎖ 농도로 DMSO로 용해한 후, 37℃에서 15분 동안 상기의 돼지 췌장 리파아제 효소와 함께 교반하였다. Pancreatic lipase inhibitory activity was measured by modifying the method performed by Kim et al. That is, porcine pancreatic lipase was dissolved in an enzyme buffer solution (10mM MOPS, 1mM EDTA, pH 6.8) at a concentration of 0.5g/200ml at 4℃, and the lysate was centrifuged at 4000rpm. , Tris buffer solution (100mM Tris-HCl, 5mM CaCl 2 , pH 7.0) and enzyme buffer were mixed with the supernatant. The catechin complex extract prepared in Example 1 (catechin: 3 types complex extract = 6:1 weight ratio) was dissolved in DMSO at 62.5, 125, 250, and 500 µg/ml concentrations, respectively, and then at 37° C. for 15 minutes. It was stirred with porcine pancreatic lipase enzyme.
그 후, 10mM 농도로 DMF에 용해한 p-NPB(p-nitrophenyl butyrate)를 첨가하여 37℃에서 35분 동안 재반응시켰다. 반응물은 진탕한 후, ELISA 리더를 이용하여 405nm에서 흡광도를 측정하였다. 항비만 의약품으로 시판 중인 오르리스타트(orlistat, tetrahydrolipstatin 성분)를 양성대조군으로 사용하였고, 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군으로 사용하였다. 췌장 리파아제 저해활성은 시료 용액의 첨가군과 무첨가군의 흡광도 감소율로 나타내었다.Thereafter, p-NPB (p-nitrophenyl butyrate) dissolved in DMF at a concentration of 10 mM was added and re-reacted at 37° C. for 35 minutes. After shaking the reaction, the absorbance was measured at 405 nm using an ELISA reader. As an anti-obesity drug, commercially available orlistat (tetrahydrolipstatin component) was used as a positive control, and the group treated with catechin alone without the three complex extracts was used as a control. Pancreatic lipase inhibitory activity was expressed as the rate of decrease in absorbance of the sample solution-added group and the non-added group.
그 결과, 본 발명의 카테킨 복합추출물은 3종 복합추출물 없이 카테킨을 단독으로 처리한 군을 대조군에 비해 모든 구간에서 췌장 리파아제 효소의 저해 활성이 증가하여 카테킨과 3종 복합추출물의 시너지 효과가 있음을 확인하였다(도 5). 또한, 본 발명의 카테킨 복합추출물은 500ug/ml 농도에서 췌장 리파아제 효소 활성을 80% 이상 저해하는 매우 우수한 효과가 있는 것을 확인할 수 있었다.As a result, the catechin complex extract of the present invention increases the inhibitory activity of pancreatic lipase enzyme in all sections compared to the control group in the group treated with catechin alone without the three complex extracts, indicating that there is a synergistic effect of the catechin and the three complex extracts. It was confirmed (Fig. 5). In addition, it was confirmed that the catechin complex extract of the present invention has a very excellent effect of inhibiting pancreatic lipase enzyme activity by 80% or more at a concentration of 500ug/ml.
다만, 오르리스타트를 처리한 양성대조군에 대비하여 본 발명의 카테킨 복합추출물의 췌장 리파아제 효소의 저해 활성이 다소 낮게 측정되었으나, 화합물 성분인 오르리스타트는 최근 위장장애, 과민증, 담즙분비 장애, 지용성 비타민 흡수억제 등의 다양한 부작용이 있는 것으로 보고되고 있다. 따라서, 천연 성분으로 구성된 본 발명의 카테킨 복합추출물은 부작용 없이 췌장 리파아제 효소의 활성을 저해하는 점에서 종래 화합물로 이루어진 항비만 약제를 대체하여 효과적으로 사용될 수 있을 것으로 예상된다.However, compared to the positive control treated with orlistat, the inhibitory activity of the pancreatic lipase enzyme of the catechin complex extract of the present invention was measured somewhat lower, but orlistat, the compound component, recently had gastrointestinal disorders, hypersensitivity, bile secretion disorder, and fat-soluble vitamin absorption. It has been reported to have various side effects such as inhibition. Therefore, the catechin complex extract of the present invention composed of natural ingredients is expected to be effectively used in place of the anti-obesity drug composed of conventional compounds in that it inhibits the activity of pancreatic lipase enzyme without side effects.
실시예 5. 카테킨 복합추출물의 세포 독성Example 5. Cytotoxicity of catechin complex extract
상기 실시예 1에서 제조한 본 발명의 카테킨 복합추출물의 3T3-L1 세포에 대한 세포독성을 확인하였다. 구체적으로, 3T3-L1 지방전구세포(preadipocyte)(ATCC CL-173, American Type Culure Collection, Manassas, USA)를 10% 우아혈청(bovine calf serum)이 함유된 DMEM을 이용하여 배양하였으며, 분화 유도 및 성숙 배지로는 10% 소태아혈청(fetal bovine serum)이 함유된 DMEM을 이용하였고, 37℃, 5% CO2 조건의 배양기에서 배양하였다. 세포 배양용 96-웰 플레이트에 합류(confluent) 상태까지 배양한 후, 실시예 1에서 제조한 카테킨 복합추출물(카테킨: 3종 복합추출물= 6:1 중량비)을 각각 62.5, 125, 250, 및 500㎍/㎖ 농도로 처리하고, 48시간 후에, MTT(tetrazolium bromide salt)를 처리하여 2시간 동안 배양시켰다. 배양 후, 모든 배양액을 제거하고 DMSO 100㎕를 분주하여 생성된 포르마진(formazin)을 녹인 뒤 마이크로플레이트 리더기(microplate reader)로 570nm(determination wave) 및 690nm(reference wave) 파장에서 흡광도를 측정하였다.The cytotoxicity of the catechin complex extract of the present invention prepared in Example 1 was confirmed for 3T3-L1 cells. Specifically, 3T3-L1 predipocytes (ATCC CL-173, American Type Culure Collection, Manassas, USA) were cultured using DMEM containing 10% bovine calf serum, inducing differentiation and DMEM containing 10% fetal bovine serum was used as the maturation medium, and cultured in an incubator at 37° C. and 5% CO 2. After culturing to a confluent state in a 96-well plate for cell culture, the catechin complex extract (catechin: 3 types complex extract = 6:1 weight ratio) prepared in Example 1 was 62.5, 125, 250, and 500, respectively. It was treated at a concentration of µg/ml, and after 48 hours, it was treated with tetrazolium bromide salt (MTT) and incubated for 2 hours. After incubation, all cultures were removed, and 100 µl of DMSO was dispensed to dissolve the generated formazin, and absorbance was measured at 570 nm (determination wave) and 690 nm (reference wave) wavelengths with a microplate reader.
그 결과, 본 발명의 카테킨 복합추출물은 모든 농도에서 95% 이상의 세포 생존률이 확인되었으며, 세포독성을 나타내지 않는 것으로 확인되었다(도 6). As a result, it was confirmed that the catechin complex extract of the present invention had a cell viability of 95% or more at all concentrations, and did not show cytotoxicity (FIG. 6).
실시예 6. 카테킨 복합추출물의 지방세포 분화 억제Example 6. Inhibition of adipocyte differentiation of catechin complex extract
본 발명의 카테킨 복합추출물의 항비만 효과를 확인하기 위해, 3T3-L1 지방전구세포(preadipocyte)(ATCC CL-173, American Type Culure Collection, Manassas, USA)를 분화시켜 지방세포 내의 지방축적 정도를 Oil-red-O 염색을 통하여 측정하였다.In order to confirm the anti-obesity effect of the catechin complex extract of the present invention, 3T3-L1 predipocytes (ATCC CL-173, American Type Culure Collection, Manassas, USA) were differentiated to reduce the degree of fat accumulation in adipocytes. -Measured through red-O staining.
3T3-L1 세포를 24웰 플레이트에 웰당 5×104 개의 세포 수로 분주한 후, 웰이 세포로 100% 채워지는 시점이 된 이후에, 2일 동안 더 유지하였다. 세포는 MDI(0.5mM 3-isobutyl-1-methylxanthine(IBMX), 1μM 덱사메타손, 1μM 인슐린)를 포함하는 10% FBS DMEM 배지로 지방세포 분화를 2일 동안 유도하였고, 배양 48시간 후, 2μM 인슐린이 함유된 10% FBS DMEM으로 이틀 동안 배양하였다. After dispensing 3T3-L1 cells into a 24-well plate at the number of 5×10 4 cells per well, the cells were further maintained for 2 days after reaching the point where the wells were 100% filled with cells. Cells were induced to differentiate adipocytes for 2 days with 10% FBS DMEM medium containing MDI (0.5 mM 3-isobutyl-1-methylxanthine (IBMX), 1 μM dexamethasone, 1 μM insulin). After 48 hours of culture, 2 μM insulin Incubated for 2 days with 10% FBS DMEM contained.
그 후, 2일 간격으로 나흘 동안 10% FBS DMEM 배양액으로 교체하였다. 지방세포 분화를 유도하는 동안 실시예 1에서 제조한 본 발명의 카테킨 복합추출물을 62.5, 125, 250, 및 500㎍/㎖ 농도로 각 배양에 처리하였고, 분화가 완성되는 시점인 8일째에 지방의 축적 정도를 확인하기 위해 Oil-red-O 염색을 실시하였다. Then, it was replaced with 10% FBS DMEM culture solution for 4 days at 2 days intervals. While inducing adipocyte differentiation, the catechin complex extract of the present invention prepared in Example 1 was treated in each culture at concentrations of 62.5, 125, 250, and 500 µg/ml. Oil-red-O staining was performed to check the degree of accumulation.
그 결과, 도 7에 나타난 바와 같이 카테킨 단일 성분에 비해 본 발명의 카테킨 복합추출물에 의한 지방축적 억제 효과가 증가하였으며, 카테킨 500ug/ml 단일처리군에서 26% 억제 활성을 보인 것에 비해, 본 발명의 카테킨 복합추출물 500ug/ml 처리군에서는 34% 수분의 지방세포 분화 억제율이 관찰되어 카테킨 단일처리군에 비해 31% 개선된 지방세포 분화 억제 효과가 관찰되었다. As a result, as shown in FIG. 7, the inhibitory effect of fat accumulation by the catechin complex extract of the present invention was increased compared to the single component of catechin, and compared to the 26% inhibitory activity in the
다만, 오르리스타트를 처리한 양성대조군에 대비하여 본 발명의 카테킨 복합추출물의 지방세포 분화 억제 활성이 다소 낮게 측정되었으나, 화합물 성분인 오르리스타트는 최근 위장장애, 과민증, 담즙분비 장애, 지용성 비타민 흡수억제 등의 다양한 부작용이 있는 것으로 보고되고 있다. 따라서, 천연 성분으로 구성된 본 발명의 카테킨 복합추출물은 부작용 없이 지방세포 분화 억제 활성을 나타내는 점에서 종래 화합물로 이루어진 항비만 약제를 대체하여 효과적으로 사용될 수 있을 것으로 예상된다.However, compared to the positive control treated with orlistat, the activity of inhibiting adipocyte differentiation of the catechin complex extract of the present invention was measured somewhat lower, but orlistat, a compound component, recently inhibited gastrointestinal disorders, hypersensitivity, bile secretion disorders, and absorption of fat-soluble vitamins. It has been reported to have various side effects such as. Therefore, it is expected that the catechin complex extract of the present invention composed of natural ingredients can be effectively used in place of anti-obesity drugs composed of conventional compounds in that it exhibits adipocyte differentiation inhibitory activity without side effects.
이상으로 본 발명의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서 이러한 구체적인 기술은 단지 바람직한 구현예일 뿐이며, 이에 본 발명의 범위가 제한되는 것이 아닌 점은 명백하다. 따라서, 본 발명의 실질적인 범위는 첨부된 청구항과 그의 등가물에 의하여 정의된다고 할 것이다.As described above, a specific part of the present invention has been described in detail, and it is obvious that this specific technology is only a preferred embodiment for those of ordinary skill in the art, and the scope of the present invention is not limited thereto. Accordingly, it will be said that the substantial scope of the present invention is defined by the appended claims and their equivalents.
Claims (7)
현미, 서목태, 및 효모의 복합추출물을 유효성분으로 포함하는,
항비만용 조성물.With catechins,
Containing a complex extract of brown rice, Seomoktae, and yeast as an active ingredient,
Anti-obesity composition.
상기 카테킨과 상기 현미, 서목태, 및 효모의 복합추출물은 6 : 0.5 내지 1.5의 중량비를 갖는 것을 특징으로 하는,
항비만용 조성물.The method of claim 1,
The complex extract of the catechin and the brown rice, Seomoktae, and yeast is characterized in that it has a weight ratio of 6: 0.5 to 1.5,
Anti-obesity composition.
상기 조성물은 항산화 활성을 갖는 것을 특징으로 하는,
항비만용 조성물.The method of claim 1,
The composition is characterized in that it has antioxidant activity,
Anti-obesity composition.
상기 조성물은 α-아밀라아제 효소 및 췌장 지방분해효소의 활성을 저해하는 것을 특징으로 하는,
항비만용 조성물.The method of claim 1,
The composition is characterized in that inhibiting the activity of α-amylase enzyme and pancreatic lipolytic enzyme,
Anti-obesity composition.
상기 조성물은 지방전구세포(preadipocyte)의 분화 억제를 통해 항비만 활성을 갖는 것을 특징으로 하는,
항비만용 조성물.The method of claim 1,
The composition is characterized in that it has anti-obesity activity through inhibition of differentiation of precipocytes (preadipocyte),
Anti-obesity composition.
상기 조성물은 약학적 조성물인 것을 특징으로 하는,
항비만용 조성물.The method according to any one of claims 1 to 5,
The composition is characterized in that the pharmaceutical composition,
Anti-obesity composition.
상기 조성물은 식품 조성물인 것을 특징으로 하는,
항비만용 조성물.
The method according to any one of claims 1 to 5,
The composition is characterized in that the food composition,
Anti-obesity composition.
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