KR20210074887A - Composition comprising sprout wheat-derived for improvement of hepatopathy - Google Patents
Composition comprising sprout wheat-derived for improvement of hepatopathy Download PDFInfo
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- KR20210074887A KR20210074887A KR1020190166021A KR20190166021A KR20210074887A KR 20210074887 A KR20210074887 A KR 20210074887A KR 1020190166021 A KR1020190166021 A KR 1020190166021A KR 20190166021 A KR20190166021 A KR 20190166021A KR 20210074887 A KR20210074887 A KR 20210074887A
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- Prior art keywords
- sprout
- extract
- composition
- oxidative stress
- wheat
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Abstract
Description
본 발명은 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 비알코올성 간 기능 손상 예방 또는 개선용 식품 조성물, 사료 조성물 및 간질환 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention relates to a food composition for preventing or improving non-alcoholic liver function damage, a feed composition, and a pharmaceutical composition for preventing or treating liver disease, comprising a sprout wheat extract or a compound derived therefrom as an active ingredient.
간은 동물의 주요 생체기관중 하나로 해독작용, 단백질의 합성, 글리코겐과 같은 양분의 저장, 지방의 대사, 담즙과 요소의 생성 및 적혈구의 분해 등 여러 역할을 수행하며, 다양한 생화학적 작용을 조절하면서 평상시 건강의 유지를 위한 기본적인 조절기능을 가지고 있다.The liver is one of the main living organs of animals, and performs various roles such as detoxification, protein synthesis, storage of nutrients such as glycogen, metabolism of fat, production of bile and urea, and decomposition of red blood cells. It has basic control functions for maintaining normal health.
간은 우리 체내에서 독소로 인식될 수 있는 각종 약물이나 술, 기타 독성물질을 분해, 대사하여 배설될 수 있는 형태로 만들어 소변이나 담즙을 통해서 배출하는 작용을 한다. 간경변증 환자에서는 이러한 해독작용이 저하되어 약물, 술, 독성물질에 의한 위험성이 증가될 수 있으므로 유의하여야 한다. 그리고 간은 중요한 면역기관이며 동시에 체내의 살균작용을 담당한다. 대장 점막을 통해서 혈액에 흡수되어 몸으로 들어간 균은 간을 거치면서 쿠퍼 세포라는 대식작용(균을 잡아먹는 기능)을 하는 세포에 의해 다 죽기 때문에 약 1% 미만의 세균만이 무사히 간을 통과해서 나갈 수 있다.The liver decomposes, metabolizes and metabolizes various drugs, alcohol, and other toxic substances that can be recognized as toxins in our body, and excretes them through urine or bile. In patients with liver cirrhosis, this detoxification effect is lowered, so the risk of drugs, alcohol, and toxic substances may increase. The liver is an important immune organ and at the same time is responsible for sterilizing the body. Bacteria that are absorbed into the blood through the large intestine mucosa and enter the body pass through the liver and die by the phagocytic cells called Kupffer cells, so only about 1% of the bacteria pass through the liver safely. can go out
한편, 인체는 산화촉진물질과 산화억제물질이 균형을 이루고 있으나, 여러 가지 요인들로 인하여 이러한 균형 상태를 잃고 산화를 촉진하는 방향으로 기울게 되면, 생체 내에 산화스트레스(oxidative stress)가 유발되어 세포손상 및 병리적 질환을 유발하게 된다. 이러한 산화적 스트레스의 직접적 원인이 되는 활성산소(reactive oxygen species, ROS)는 화학적으로 불안정하고 반응성이 높아 DNA, 단백질, 지질 및 탄수화물과 같은 여러 생체물질과 쉽게 반응할 수 있으며, 생체 내 고분자들을 공격하여 세포와 조직에 비가역적인 손상을 일으키거나 돌연변이, 세포독성 및 암 등을 초래하게 된다. 실제로 많은 임상 질환에서 산화스트레스가 증가되는 것이 확인되었으며, 이러한 산화스트레스를 줄이기 위해 새로운 항산화 물질에 대한 다양한 연구가 이루어지고 있다.On the other hand, the human body has a balance between the oxidation-promoting substance and the oxidation-inhibiting substance. However, if the balance is lost due to various factors and the body tilts in the direction to promote oxidation, oxidative stress is induced in the living body, causing cell damage. and pathological diseases. Reactive oxygen species (ROS), which is a direct cause of oxidative stress, are chemically unstable and highly reactive, so they can easily react with various biomaterials such as DNA, proteins, lipids and carbohydrates, and attack macromolecules in the living body. This leads to irreversible damage to cells and tissues, mutation, cytotoxicity, and cancer. In fact, it has been confirmed that oxidative stress is increased in many clinical diseases, and various studies are being made on new antioxidants to reduce this oxidative stress.
특히, 천연물을 대상으로 항산화 물질을 검색하여, 항산화 활성과 안전성을 동시에 도모하려는 연구가 활발히 진행되고 있다. 예를 들어, 한국특허등록 제1022705호에는 항산화 활성효능을 가진 감태(Ecklonia cava)의 에탄올 추출물이 개시되어 있다. 그러나 아직까지는 항산화 활성과 안전성을 동시에 도모할 수 있는 천연물에 대한 연구결과가 많이 보고되고 있지 않아, 추가적인 연구가 필요한 실정이다.In particular, research is being actively conducted to search for antioxidants in natural products to promote antioxidant activity and safety at the same time. For example, Korean Patent Registration No. 1022705 discloses an ethanol extract of Ecklonia cava having antioxidant activity. However, research results on natural products that can promote antioxidant activity and safety at the same time have not been reported yet, so additional research is needed.
상기와 같은 배경 하에, 본 발명자들은 산화스트레스로부터의 간 기능 개선에 대하여 연구하던 중, 새싹 밀 추출물 또는 이로부터 유래한 화합물 이소오리엔틴의 산화스트레스로부터의 간세포 보호 및 간 기능 개선 효과를 확인함으로써, 본 발명을 완성하였다.Under the above background, the present inventors, while studying for improvement of liver function from oxidative stress, by confirming the hepatocellular protection and liver function improvement effect of sprout wheat extract or a compound isoorientin derived therefrom from oxidative stress, The present invention was completed.
본 발명의 하나의 목적은 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 비알코올성 간 기능 손상 예방 또는 개선용 식품 조성물을 제공하는 것이다.One object of the present invention is to provide a food composition for preventing or improving non-alcoholic liver function damage, comprising a sprout wheat extract or a compound derived therefrom as an active ingredient.
본 발명의 다른 목적은 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 비알코올성 간 기능 손상 예방 또는 개선용 사료 조성물을 제공하는 것이다.Another object of the present invention is to provide a feed composition for preventing or improving non-alcoholic liver function damage, comprising a wheat sprout extract or a compound derived therefrom as an active ingredient.
본 발명의 또 다른 목적은 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 간질환 예방 또는 치료용 약학 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating liver disease comprising a wheat sprout extract or a compound derived therefrom as an active ingredient.
상기 목적을 달성하기 위한 본 발명의 일 실시양태는 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 비알코올성 간 기능 손상 예방 또는 개선용 식품 조성물을 제공한다.One embodiment of the present invention for achieving the above object provides a food composition for preventing or improving non-alcoholic liver function damage comprising a wheat sprout extract or a compound derived therefrom as an active ingredient.
구체적으로, 상기 화합물은 이소오리엔틴인 것일 수 있으며, 상기 비알코올성 간 기능 손상은 산화스트레스에 의한 것일 수 있고, 상기 조성물은 산화스트레스로부터 간세포를 보호하는 것일 수 있으며, 상기 보호는 활성산소 생성을 억제하고, 글루타티온을 증가시키며, 항산화 효소의 활성을 조절하는 것일 수 있고, 보다 구체적으로, 상기 항산화 효소는 카탈라아제(Catalase; CAT), 글루타티온 환원효소(Glutathione reductase; GR) 및 글루타티온 과산화효소(Glutathione peroxidase;GPx)일 수 있으나, 이에 제한되지 않는다.Specifically, the compound may be isoorientin, the non-alcoholic liver function impairment may be due to oxidative stress, the composition may be to protect the liver cells from oxidative stress, and the protection may result in the generation of active oxygen Inhibits, increases glutathione, and modulates the activity of antioxidant enzymes, and more specifically, the antioxidant enzymes are catalase (CAT), glutathione reductase (GR) and glutathione peroxidase ;GPx), but is not limited thereto.
본 발명의 용어, “새싹 밀”은 밀에 싹을 틔워 약 15cm정도 자란 어린 식물체를 의미한다. 통상적으로, 상기 새싹 밀은 밀을 하루정도 물에 침지한 후, 수분을 제거하고, 수분의 손실을 방지한 조건에서 파종하여 발아시킨 다음, 3 내지 20일 동안 성장시켜서 수득할 수 있다.As used herein, the term "sprout wheat" refers to a young plant that has sprouted on wheat and grown to about 15 cm. Typically, the sprout wheat can be obtained by immersing wheat in water for about a day, removing moisture, sown under conditions that prevent moisture loss, germinate, and then grow for 3 to 20 days.
본 발명에 있어서, 상기 새싹 밀의 추출물은 산화스트레스에 의한 간 손상을 예방, 치료 또는 개선하는데 사용될 수 있다.In the present invention, the extract of wheat sprout can be used to prevent, treat or improve liver damage caused by oxidative stress.
본 발명의 용어, “추출물”은 목적하는 물질을 다양한 용매에 침지한 다음, 상온 또는 가온상태에서 일정시간 동안 추출하여 수득한 액상성분, 상기 액상성분으로부터 용매를 제거하여 수득한 고형분 등의 결과물을 의미한다. 뿐만 아니라, 상기 결과물에 더하여, 상기 결과물의 희석액, 이들의 농축액, 이들의 조정제물, 정제물 등을 모두 포함하는 것으로 포괄적으로 해석될 수 있다.As used herein, the term “extract” refers to a liquid component obtained by immersing a target substance in various solvents and then extracting it at room temperature or in a heated state for a certain time, and a solid obtained by removing the solvent from the liquid component. it means. In addition, in addition to the result, it can be comprehensively interpreted as including all of the diluent of the result, the concentrate thereof, the prepared product thereof, the purified product, and the like.
본 발명에 있어서, 상기 추출물은 새싹 밀 추출물을 의미한다. 상기 새싹 밀 추출물은 상기 새싹밀의 식물전체, 이의 건조물, 가공물 등을 단독으로 또는 조합하여, 물 또는 다양한 유기용매 등으로 추출하여 수득할 수 있다.In the present invention, the extract means sprout wheat extract. The sprout wheat extract can be obtained by extracting the whole plant of the sprout wheat, its dried products, processed products, etc., alone or in combination, with water or various organic solvents.
이때, 사용되는 유기용매는 특별히 이에 제한되지 않으나, 바람직하게는 물, 극성용매 또는 비극성용매가 될 수 있고, 보다 바람직하게는 물, 탄소수 1 내지 4의 저급 알코올(메탄올, 에탄올, 프로판올 또는 부탄올 등), 이들의 혼합용매 등이 될 수 있으며, 가장 바람직하게는 에탄올 또는 그의 혼합용매를 사용할 수 있다. 또한, 상기 추출물을 수득하기 위한 방법 역시 특별히 이에 제한되지 않으나, 바람직하게는 상기 새싹밀의 식물전체, 이의 건조물, 가공물 등을 상기 용매에 침지하고, 10 내지 25℃의 상온에서 추출하는 냉침추출법, 40 내지 100℃로 가열하여 추출하는 가열추출법, 초음파를 가하여 추출하는 초음파추출법, 환류냉각기를 이용한 환류추출법 등의 방법을 사용할 수 있다.At this time, the organic solvent used is not particularly limited thereto, but preferably water, a polar solvent or a non-polar solvent, and more preferably water, a lower alcohol having 1 to 4 carbon atoms (methanol, ethanol, propanol or butanol, etc.) ), and a mixed solvent thereof, and most preferably, ethanol or a mixed solvent thereof may be used. In addition, the method for obtaining the extract is also not particularly limited thereto, but preferably the whole plant of the sprout wheat, its dried product, processed product, etc. is immersed in the solvent, and extracted at room temperature of 10 to 25 ° C. Cold extraction method, 40 Methods such as a heating extraction method in which extraction is performed by heating to 100° C., an ultrasonic extraction method in which ultrasonic waves are applied, and a reflux extraction method using a reflux condenser can be used.
본 발명에 있어서, 상기 추출물의 새싹 밀 함량은 특별히 이에 제한되지 않으나, 최종 조성물 총 중량을 기준으로 0.0001 내지 80 중량%, 0.0001 내지 50 중량%, 보다 구체적으로 0.01 내지 20 중량%의 함량으로 포함될 수 있다.In the present invention, the sprout wheat content of the extract is not particularly limited thereto, but may be included in an amount of 0.0001 to 80% by weight, 0.0001 to 50% by weight, more specifically 0.01 to 20% by weight, based on the total weight of the final composition. have.
본 발명에 있어서, 상기 화합물은 “이소오리엔틴(isoorientin)”일 수 있다.In the present invention, the compound may be "isoorientin".
상기 용어, “이소오리엔틴”은 새싹 밀로부터 유래되고, 하기 화학식 1로 표시되는 화합물을 의미한다. 상기 이소오리엔틴은 새싹 밀과 같은 천연 공급원으로부터 분리할 수 있으나, 이에 한정되지 않고, 당 업계에 공지된 방법으로 화학적으로 합성하거나, 시판되는 물질을 사용할 수 있다.The term, “isoorientin” is derived from sprout wheat and refers to a compound represented by the following formula (1). The isoorientin may be isolated from a natural source such as sprout wheat, but is not limited thereto, and a commercially available material may be used or chemically synthesized by a method known in the art.
본 발명에 있어서, 상기 이소오리엔틴의 함량은 특별히 이에 제한되는 것은 아니나, 최종 조성물 총 중량을 기준으로 0.0001 내지 80 중량%, 0.0001 내지 50 중량%, 보다 구체적으로 0.01 내지 20 중량%의 함량으로 포함될 수 있다.In the present invention, the content of isoorientin is not particularly limited thereto, but based on the total weight of the final composition, 0.0001 to 80% by weight, 0.0001 to 50% by weight, more specifically 0.01 to 20% by weight. can
본 발명의 용어, “간”은 우리 체내에서 독소로 인식될 수 있는 각종 약물이나 술, 기타 독성물질을 분해, 대사하여 배설될 수 있는 형태로 만들어 소변이나 담즙을 통해서 배출하는 작용을 한다. 간경변증 환자에서는 이러한 해독작용이 저하되어 약물, 술, 독성물질에 의한 위험성이 증가될 수 있으므로 유의하여야 한다. 그리고 간은 중요한 면역기관이며 동시에 체내의 살균작용을 담당한다. 대장 점막을 통해서 혈액에 흡수되어 몸으로 들어간 균은 간을 거치면서 쿠퍼 세포라는 대식작용(균을 잡아먹는 기능)을 하는 세포에 의해 다 죽기 때문에 약 1% 미만의 세균만이 무사히 간을 통과해서 나갈 수 있다.As used herein, the term "liver" decomposes, metabolizes and metabolizes various drugs, alcohol, and other toxic substances that can be recognized as toxins in our body and excretes them through urine or bile. In patients with liver cirrhosis, this detoxification effect is lowered, so the risk of drugs, alcohol, and toxic substances may increase. The liver is an important immune organ and at the same time is responsible for sterilizing the body. Bacteria that are absorbed into the blood through the large intestine mucosa and enter the body pass through the liver and die by the phagocytic cells called Kupffer cells, so only about 1% of the bacteria pass through the liver safely. can go out
상기 용어, “산화스트레스”는 인체의 에너지 생성과정(섭취한 음식물이 산소와 반응하는 산화과정을 거침) 중에 유해산소가 급격히 증가해 인체에 나쁜 영향을 일으키거나 그러한 상황을 의미한다.The term “oxidative stress” refers to a situation in which harmful oxygen is rapidly increased during the energy generation process of the human body (the food consumed undergoes an oxidation process that reacts with oxygen), causing a bad effect on the human body or such a situation.
본 발명의 용어, "예방"은 본 발명에 따른 약학적 조성물에 의해 간 기능의 손상을 억제시키거나 또는 지연시키는 모든 행위를 의미한다.As used herein, the term “prevention” refers to any act of inhibiting or delaying damage to liver function by the pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어, "개선"은 본 발명의 조성물을 이용하여 간 기능의 손상을 억제 또는 지연시키거나, 이미 손상된 간 기능을 완화시키는 것을 말한다.As used herein, the term "improvement" refers to inhibiting or delaying damage to liver function or alleviating liver function already damaged by using the composition of the present invention.
본 발명의 용어, “식품”이란, 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올음료, 비타민 복합제, 건강기능 식품 및 건강식품 등이 있으며, 통상적인 의미에서의 식품을 모두 포함한다.As used herein, the term “food” refers to meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, various soups, beverages, tea, drinks, alcohol There are beverages, vitamin complexes, health functional foods, and health foods, and includes all foods in the ordinary sense.
본 발명의 식품 조성물은, 일상적으로 섭취하는 것이 가능하기 때문에 우수한 비알코올성 간 기능 개선 효과를 기대할 수 있으므로, 건강 증진 목적으로 매우 유용하게 사용될 수 있다.Since the food composition of the present invention can be consumed on a daily basis, an excellent non-alcoholic liver function improvement effect can be expected, and thus it can be very usefully used for health promotion purposes.
본 발명의 비알코올성 간 기능 손상 예방 또는 개선 효과를 나타낼 수 있다면 식품 조성물에 다양한 중량%로 포함될 수 있다. 구체적으로 식품 조성물의 총 중량 대비 0.00001 내지 100 중량% 또는 0.01 내지 80 중량%로 포함될 수 있으나, 이에 제한되지 않는다. 건강 및 위생을 목적으로 장기간 섭취할 경우에는 상기 범위 이하의 함량을 포함할 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다.If the non-alcoholic liver function damage prevention or improvement effect of the present invention can be exhibited, it may be included in various weight % in the food composition. Specifically, it may be included in an amount of 0.00001 to 100% by weight or 0.01 to 80% by weight relative to the total weight of the food composition, but is not limited thereto. When ingested for a long period of time for health and hygiene purposes, it may contain an amount below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.
상기 용어, “활성산소(Reactive oxygen species; ROS)”는 세포 내의 미토콘드리아에서 호흡과 면역 반응에 의한 산소의 산화와 환원 과정을 통해 끊임없이 생성된다. 유해한 ROS는 일반적으로 슈퍼옥사이드 디스뮤타제(superoxide dismutase), 카탈라아제(catalase), 글루타티온 과산화효소(glutathione peroxidase), 글루타민 환원효소(glutamine reductase), 비타민 C, 비타민 E, 요산, 빌리루빈(bilirubin)과 같은 항산화 체계의 작용으로 제거된다. 그러나 자유기의 생성과 소거의 균형이 무너지게 되면 산화적 스트레스(oxidative stress)가 발생하게 되어 염증, 노화 또는 암과 같은 다양한 병리학적인 변화를 일으킨다. 실제로 많은 임상 질환에서 산화스트레스가 증가되는 것이 확인되었으며, 이러한 산화스트레스를 줄이기 위해 새로운 항산화 물질에 대한 다양한 연구가 이루어지고 있다.The term, “reactive oxygen species (ROS)” is constantly generated through the oxidation and reduction processes of oxygen by respiration and immune responses in mitochondria in cells. Harmful ROS are usually caused by superoxide dismutase, catalase, glutathione peroxidase, glutamine reductase, vitamin C, vitamin E, uric acid, and bilirubin. It is eliminated by the action of the antioxidant system. However, when the balance between generation and elimination of free radicals is disrupted, oxidative stress occurs, causing various pathological changes such as inflammation, aging, or cancer. In fact, it has been confirmed that oxidative stress is increased in many clinical diseases, and various studies are being made on new antioxidants to reduce this oxidative stress.
상기 용어, “글루타티온(Glutathione; GSH)”은 식물, 동물 그리고 일부 세균(bacteria)에서 매우 중요한 항산화제인데, 글루탐산(glutamate), 시스테인(cysteine), 글라이신(glycine) 세 개의 펩타이드로 구성되어 있다고 알려져 있다.The term, “Glutathione (GSH)” is a very important antioxidant in plants, animals, and some bacteria. It is known that it is composed of three peptides: glutamate, cysteine, and glycine. have.
상기 용어, “카탈라아제(Catalase; CAT)”는 과산화수소가 분해되어 물과 산소가 만들어지는 반응을 촉매하는 효소이며, 우리 몸속의 간, 적혈구, 신장에 들어 있다고 알려져 있다.The term, “catalase (CAT)” is an enzyme that catalyzes a reaction in which hydrogen peroxide is decomposed to produce water and oxygen, and is known to be contained in the liver, red blood cells, and kidneys in our body.
상기 용어, “글루타티온 환원효소(Glutathione reductase; GR)”는 산화형 글루타티온(GS-SD)을 환원형 NAD 또는 NADP에 의해 환원형(GSH)로 만드는 반응을 돕는 효소를 의미한다.As used herein, the term “glutathione reductase (GR)” refers to an enzyme that helps in the reaction of converting oxidized glutathione (GS-SD) to reduced form (GSH) by reduced NAD or NADP.
상기 용어, “글루타티온 과산화효소(Glutathione peroxidase;GPx)”는 글루타티온과 과산화수소 또는 지질과산화물로부터 산화형 글루타티온과 물 또는 알코올을 생성하는 반응을 촉매하는 효소를 의미한다. 구체적으로, 과산화효소의 일종으로 생체 내에서 과산화수소나 지질과산화물을 제거하는 작용을 한다고 알려져 있다.As used herein, the term “glutathione peroxidase (GPx)” refers to an enzyme that catalyzes a reaction between glutathione and hydrogen peroxide or lipid peroxide to produce oxidized glutathione and water or alcohol. Specifically, it is a type of peroxidase and is known to act to remove hydrogen peroxide or lipid peroxide in a living body.
상기 목적을 달성하기 위한 본 발명의 다른 실시 양태는 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 비알코올성 간 기능 손상 예방 또는 개선용 사료 조성물을 제공한다.Another embodiment of the present invention for achieving the above object provides a feed composition for preventing or improving non-alcoholic liver function damage comprising a sprout wheat extract or a compound derived therefrom as an active ingredient.
상기 용어, “새싹 밀”, “추출물”, “화합물”, “간”, “예방” 및 “개선”은 상기에서 서술한 바와 같다.The terms “sprout wheat”, “extract”, “compound”, “liver”, “prevention” and “improvement” are as described above.
본 발명의 용어, “사료”는 동물이 먹고, 섭취하며, 소화시키기 위한 또는 이에 적당한 임의의 천연 또는 인공 규정식, 한끼식 등 또는 상기 한끼식의 성분을 의미한다.As used herein, the term “feed” means any natural or artificial diet, meal meal, etc., or a component of the meal meal, intended for or suitable for being eaten, ingested, and digested by an animal.
상기 사료의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용되는 사료를 사용할 수 있다. 상기 사료의 비제한적인 예로는, 곡물류, 근과류, 식품 가공 부산물류, 조류, 섬유질류, 제약 부산물류, 유지류, 전분류, 박류 또는 곡물 부산물류 등과 같은 식물성 사료; 단백질류, 무기물류, 유지류, 광물성류, 유지류, 단세포 단백질류, 동물성 플랑크톤류 또는 음식물 등과 같은 동물성 사료를 들 수 있다. 이들은 단독으로 사용되거나 2종 이상을 혼합하여 사용될 수 있다.The type of feed is not particularly limited, and feed commonly used in the art may be used. Non-limiting examples of the feed include plant feeds such as grains, root fruits, food processing by-products, algae, fibers, pharmaceutical by-products, oils and fats, starches, gourds or grain by-products; and animal feeds such as proteins, inorganic materials, oils and fats, minerals, oils and fats, single cell proteins, zooplankton or food. These may be used alone or in mixture of two or more.
상기 목적을 달성하기 위한 본 발명의 또 다른 실시 양태는 새싹 밀 추출물 또는 이로부터 유래된 화합물을 유효성분으로 포함하는 간질환의 예방 또는 치료용 약학 조성물을 제공한다.Another embodiment of the present invention for achieving the above object provides a pharmaceutical composition for the prevention or treatment of liver disease comprising a wheat sprout extract or a compound derived therefrom as an active ingredient.
상기 용어, “새싹 밀”, “추출물”, “화합물”, “간” 및 “예방”은 상기에서 서술한 바와 같다.The terms “sprout wheat”, “extract”, “compound”, “liver” and “prevention” are as described above.
본 발명의 용어, "치료"는 상기 약학적 조성물에 의해 산화스트레스에 의해 간 기능이 손상된 개체의 증상이 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used herein, the term “treatment” refers to any action in which the symptoms of an individual whose liver function is impaired by oxidative stress by the pharmaceutical composition is improved or is beneficially changed.
본 발명의 약학적 조성물은, 약학적으로 허용 가능한 담체, 부형제 또는 희석제를 추가로 포함할 수 있는데, 상기 담체는 비자연적 담체(non-naturally occuring carrier)를 포함할 수 있다.The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient or diluent, which may include a non-naturally occurring carrier.
구체적으로, “약학적으로 허용가능한 담체”는 생물체를 자극하지 않고 본 발명 조성물의 피부과색소 침착성 질환 증상의 예방 또는 치료 활성 및 특성을 저해하지 않는 담체 또는 희석제를 말한다. 액상 용액으로 제제화되는 조성물에 있어서 허용되는 약학적 담체로는, 멸균, 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 하나 이상의 성분을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다.Specifically, "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not stimulate the organism and does not inhibit the preventive or therapeutic activity and properties of the composition of the present invention for preventing or treating hyperpigmentation disease symptoms. Examples of acceptable pharmaceutical carriers for compositions formulated as liquid solutions are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection, dextrose solution, maltodextrin solution, glycerol, ethanol. And one or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostats may be added as necessary.
또한, 본 발명의 약학적으로 허용가능한 담체는 비자연적 담체를 포함할 수 있다.In addition, the pharmaceutically acceptable carrier of the present invention may include a non-natural carrier.
보다 구체적으로, 상기 약학적 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트, 폴리카프로락톤(polycaprolactone), 폴리락틱액시드(Poly Lactic Acid), 폴리-L-락틱액시드(poly-L-lactic acid), 광물유 등을 들 수 있다.More specifically, carriers, excipients and diluents that may be included in the pharmaceutical composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate , calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, polycaprolactone, polylactic acid (Poly Lactic Acid), poly-L-lactic acid, mineral oil, and the like.
상기 약학적 조성물은 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있으며, 담체의 형태로는 각종 부정형의 담체, 마이크로 스피어, 나노파이버 등을 포함할 수 있다.The pharmaceutical composition may be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., oral dosage forms, external preparations, suppositories, and sterile injection solutions according to conventional methods, respectively. The carrier may include various amorphous carriers, microspheres, nanofibers, and the like.
제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다.In the case of formulation, it can be prepared using a diluent or excipient such as a filler, extender, binder, wetting agent, disintegrant, surfactant, etc. commonly used.
경구투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 상기 추출물과 이의 분획물들에 적어도 하나 이상의 부형제 예를 들면, 전분, 칼슘 카보네이트(calcium carbonate), 수크로스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 마그네슘 스티레이트, 탈크 같은 윤활제들도 사용될 수 있다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and these solid preparations include at least one excipient in the extract and its fractions, for example, starch, calcium carbonate, It may be prepared by mixing sucrose or lactose, gelatin, or the like. In addition to simple excipients, lubricants such as magnesium stearate and talc may also be used.
경구투여를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다.Liquid formulations for oral administration include suspensions, solutions, emulsions, syrups, etc. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have.
비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제 등이 포함될 수 있다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다.Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate.
본 발명의 약학적 조성물은 약학적으로 유효한 양으로 투여될 수 있는데, 상기 용어, “약학적으로 유효한 양”은 의학적 치료 또는 예방에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료 또는 예방하기에 충분한 양을 의미하며, 유효 용량 수준은 질환의 중증도, 약물의 활성, 환자의 연령, 체중, 건강, 성별, 환자의 약물에 대한 민감도, 사용된 본 발명 조성물의 투여 시간, 투여 경로 및 배출 비율 치료기간, 사용된 본 발명의 조성물과 배합 또는 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 약학적 조성물은 단독으로 투여하거나 공지된 익상편 치료제와 병용하여 투여될 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하다.The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount, and the term, “pharmaceutically effective amount” is sufficient to treat or prevent a disease at a reasonable benefit/risk ratio applicable to medical treatment or prevention. means the amount, and the effective dose level is the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the administration time of the composition of the present invention used, the route of administration and the rate of excretion Treatment period , factors including drugs used in combination with or concurrently with the composition of the present invention and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a known therapeutic agent for pterygium. Taking all of the above factors into consideration, it is important to administer an amount capable of obtaining the maximum effect with a minimum amount without side effects.
본 발명에서 사용되는 용어, "투여"는 어떠한 적절한 방법으로 환자에게 본 발명의 조성물 도입하는 것을 의미하며, 상기 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 투여될 수 있다. 복강 내 투여, 정맥 내 투여, 근육 내 투여, 피하 투여, 피내 투여, 경구 투여, 국소 투여, 비 내 투여될 수 있으나, 이에 제한되지는 않는다. 본 발명의 조성물이 항산화, 미백, 피부 노화 또는 주름 개선에 효과가 있는 특성상, 상기 조성물의 투여 경로는 피부에 도포하여 투여될 수 있다.As used herein, the term "administration" means introducing the composition of the present invention to a patient by any suitable method, and the administration route of the composition may be administered through any general route as long as it can reach the target tissue. . Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, may be administered intranasally, but is not limited thereto. Since the composition of the present invention is effective in antioxidant, whitening, skin aging or wrinkle improvement, the route of administration of the composition may be applied to the skin and administered.
본 발명의 조성물을 매일 투여 또는 간헐적으로 투여해도 좋고, 1일당 투여 횟수는 1회 또는 2 ~ 3회로 나누어 투여하는 것이 가능하다. 또한, 본 발명의 조성물은 항산화, 피부 노화 또는 주름 개선 위하여 단독으로, 또는 다른 약물 치료와 병용하여 사용할 수 있다. 상기 요소를 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 당업자에 의해 용이하게 결정될 수 있다.The composition of the present invention may be administered daily or intermittently, and the number of administrations per day may be administered once or divided into two to three times. In addition, the composition of the present invention may be used alone or in combination with other drug treatments for antioxidation, skin aging or wrinkle improvement. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, and can be easily determined by those skilled in the art.
본 발명에서 용어, "개체"는 주름 또는 피부 노화가 발병하였거나 발병할 수 있는 인간을 포함한 쥐, 생쥐, 가축 등의 모든 동물을 의미한다. 구체적으로, 인간을 포함한 포유동물일 수 있다.As used herein, the term "individual" refers to all animals, such as rats, mice, and livestock, including humans, that have or may develop wrinkles or skin aging. Specifically, it may be a mammal including a human.
본 발명에 있어서, 상기 “간질환”은 간장(肝臟)의 기능에 장애가 생기는 것으로서, 구체적으로, 상기 간질환은 간 기능 손상에 의한 것으로, 이는 산화스트레스에 의한 것일 수 있으며, 보다 구체적으로, 상기 간질환은 비알코올성 간 손상인 것일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the "liver disease" refers to a disorder in the function of the liver, and specifically, the liver disease is caused by impaired liver function, which may be due to oxidative stress, and more specifically, the Liver disease may be non-alcoholic liver damage, but is not limited thereto.
본 발명에 있어서, 상기 조성물은 산화스트레스로부터 간세포를 보호하는 것이며, 구체적으로 상기 보호는 활성산소 생성 억제에 의한 것일 수 있고, 글루타티온 증가에 의한 것일 수 있으며, 항산화 효소의 활성 조절에 의한 것일 수 있고, 보다 구체적으로, 상기 항산화 효소는 카탈라아제, 글루타티온 환원효소 또는 글루타티온 과산화효소인 것일 수 있으나, 이에 제한되는 것은 아니다.In the present invention, the composition protects hepatocytes from oxidative stress, and specifically, the protection may be by inhibition of active oxygen production, by increase in glutathione, or by regulation of the activity of antioxidant enzymes, , More specifically, the antioxidant enzyme may be catalase, glutathione reductase or glutathione peroxidase, but is not limited thereto.
본 발명에 따른 새싹밀 추출물 또는 이의 화합물을 유효성분으로 포함하는 조성물은, 산화스트레스로 인한 간질환 또는 비알코올성 간 기능 손상을 예방, 치료 또는 개선할 수 있는 효과가 있다.The composition comprising the sprout wheat extract or a compound thereof according to the present invention as an active ingredient has an effect of preventing, treating or improving liver disease or non-alcoholic liver function damage due to oxidative stress.
도 1은 새싹 밀 추출물의 세포 독성을 나타내는 그래프이다.
도 2는 새싹 밀 추출물의 산화스트레스에 대한 보호 효과를 나타내는 그래프이다.
도 3은 새싹 밀 추출물의 ROS 생성 억제 효과를 나타내는 그래프이다.
도 4는 새싹 밀 추출물의 HepG2 세포 내 글루타티온 함량 조절을 나타내는 그래프이다.
도 5는 새싹 밀 추출물의 catalase 수준 조절을 나타내는 그래프이다.
도 6은 새싹 밀 추출물의 glutathione reductase 수준 조절을 나타내는 그래프이다.
도 7은 새싹 밀 추출물의 glutathione peroxidase 수준 조절을 나타내는 그래프이다.
도 8은 새싹 밀 추출물에서 분리한 화합물을 나타내는 분석 크로마토그램 그래프이다.
도 9는 새싹 밀 유래 화합물 이소오리엔틴의 세포 독성을 나타내는 그래프이다.
도 10은 새싹 밀 유래 화합물 이소오리엔틴의 산화스트레스에 대한 보호 효과를 나타내는 그래프이다.1 is a graph showing the cytotoxicity of sprout wheat extract.
2 is a graph showing the protective effect on oxidative stress of sprout wheat extract.
3 is a graph showing the ROS generation inhibitory effect of sprout wheat extract.
4 is a graph showing the control of glutathione content in HepG2 cells of sprout wheat extract.
5 is a graph showing the catalase level regulation of sprout wheat extract.
6 is a graph showing the control of glutathione reductase levels of wheat sprout extract.
7 is a graph showing the control of glutathione peroxidase level of wheat sprout extract.
8 is an analytical chromatogram graph showing compounds isolated from sprout wheat extract.
9 is a graph showing the cytotoxicity of the compound isoorientin derived from sprout wheat.
10 is a graph showing the protective effect on oxidative stress of the compound isoorientin derived from sprout wheat.
이하, 하기 실시예에 의하여 본 발명을 더욱 상세하게 설명하고자 한다. 단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐 본 발명의 범위가 이들만으로 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail by way of Examples. However, the following examples are only for illustrating the present invention, and the scope of the present invention is not limited thereto.
실시예 1. 새싹 밀 추출물 제조방법Example 1. Method for preparing sprout wheat extract
본 발명에 사용된 새싹 밀은 2018년도에 전북 완주 소재의 농촌진흥청 국립식량과학원 시험포장에서 재배하여 시험용 재료로 사용하였다. 추출용매는 물과 주정을 혼합하여 사용하였고, 칭량된 새싹 밀의 20배수의 용매로 24시간 1차 추출하여 추출물을 얻고 남은 잔여물을 다시 10배수의 용매로 24시간 2차 추출하였다. 용매는 감압 농축기(N-1000, EYELA, Tokyo, Japan)를 이용하여 완전히 제거하였고, 잔사는 DMSO로 다시 용해하였다. 각 추출물은 질소 충진 후 -20℃에서 보관하면서 실험에 사용하였다.Sprout wheat used in the present invention was grown in the test field of the National Institute of Food Science, Rural Development Administration, Wanju, Jeollabuk-do in 2018 and used as a test material. As the extraction solvent, a mixture of water and alcohol was used, and the extract was obtained by primary extraction for 24 hours with 20 times the solvent of the weighed sprout wheat, and the remaining residue was extracted again for 24 hours with 10 times the solvent. The solvent was completely removed using a vacuum concentrator (N-1000, EYELA, Tokyo, Japan), and the residue was dissolved again with DMSO. Each extract was used in the experiment while being stored at -20°C after filling with nitrogen.
실시예 2. 새싹 밀 추출물의 세포 독성 평가Example 2. Cytotoxicity evaluation of sprout wheat extract
인체유래 간세포인 HepG2 세포를 이용하여, 새싹 밀 추출물의 세포 독성을 측정하였다.Cytotoxicity of sprout wheat extract was measured using HepG2 cells, which are human hepatocytes.
보다 구체적으로, HepG2 세포는 ATCC(American Type Culture Collection)에서 구입하여 사용하였으며 10% FBS, 100 units/mL penicillin/streptomycin이 함유된 DMEM 배지를 사용하여 37℃, 5% CO₂로 조절된 incubator에서 배양하였다. 이후, 새싹 밀 추출물의 세포 독성을 MTT assay 방법을 이용하여 측정하였다.More specifically, HepG2 cells were purchased from the American Type Culture Collection (ATCC) and cultured in an incubator conditioned at 37°C and 5% CO₂ using DMEM medium containing 10% FBS and 100 units/mL penicillin/streptomycin. did. Then, the cytotoxicity of the sprout wheat extract was measured using the MTT assay method.
그 결과, 도 1에서 볼 수 있듯이, HepG2 세포에서 새싹 밀 추출물은 50 μg/ml 이하에서 모두 세포 독성을 나타내지 않았다.As a result, as can be seen in FIG. 1 , the sprout wheat extract in HepG2 cells did not show any cytotoxicity at 50 μg/ml or less.
이를 통해, 본 발명의 새싹 밀 추출물이 세포에 독성을 띄지 않는 것을 확인하였다.Through this, it was confirmed that the sprout wheat extract of the present invention is not toxic to cells.
실시예 3. 새싹 밀 추출물의 간세포 내 산화스트레스 보호 효과Example 3. Oxidative stress protective effect in hepatocytes of sprout wheat extract
실시예 3-1. MTT 어세이Example 3-1. MTT assay
인체유래 간세포인 HepG2 세포를 이용하여, 산화스트레스에 의한 세포 생존율을 측정하고, 새싹 밀 추출물에 의한 산화스트레스로부터의 세포 보호 효과를 확인하였다.Cell viability by oxidative stress was measured using HepG2 cells, which are human hepatocytes, and the cytoprotective effect of wheat sprout extract from oxidative stress was confirmed.
구체적으로, 세포 배양은 실시예 2와 동일하게 배양하였다. 또한, TBHP를 1mM 처리하여 산화스트레스를 유도하고, 새싹 밀 추출물의 산화스트레스에 대한 보호효과를 MTT assay 방법을 이용하여 측정하였다.Specifically, the cell culture was cultured in the same manner as in Example 2. In addition, TBHP was treated with 1 mM to induce oxidative stress, and the protective effect of sprout wheat extract against oxidative stress was measured using the MTT assay method.
그 결과, 도 2에서 볼 수 있듯이, HepG2 세포에서 산화적 스트레스를 유도하는 TBHP 처리에 의해 감소된 세포 생존율이 새싹 밀 추출물 처리 시 농도 의존적으로 세포 보호효과를 나타냄을 확인하였다.As a result, as shown in FIG. 2 , it was confirmed that the cell viability reduced by TBHP treatment inducing oxidative stress in HepG2 cells exhibited a concentration-dependent cytoprotective effect upon treatment with sprout wheat extract.
이를 통해, 새싹 밀 추출물이 산화스트레스로부터 간세포를 보호하는 효능이 있음을 확인하였다.Through this, it was confirmed that the sprout wheat extract has the effect of protecting hepatocytes from oxidative stress.
실시예 3-2. DCFH-DA 어세이Example 3-2. DCFH-DA assay
인체유래 간세포인 HepG2 세포를 이용하여, 산화스트레스에 의한 활성산소(Reactive oxygen species; ROS) 생성 억제 효과를 측정하였다.Using HepG2 cells, which are human hepatocytes, the inhibitory effect of reactive oxygen species (ROS) production by oxidative stress was measured.
구체적으로, 실시예 3-1과 동일하게 실험한 뒤, HepG2 세포에 대한 새싹 밀 추출물의 ROS 생성 억제 효과는 DCFH-DA assay 방법을 이용하였다.Specifically, after the same experiment as in Example 3-1, the ROS generation inhibitory effect of the sprout wheat extract on HepG2 cells was performed using the DCFH-DA assay method.
그 결과, 도 3에서 볼 수 있듯이, TBHP 처리에 의해 증가된 ROS 생성이 새싹 밀 추출물에 의해 농도의존적으로 억제됨을 확인하였다.As a result, as shown in FIG. 3 , it was confirmed that ROS production increased by TBHP treatment was inhibited in a concentration-dependent manner by the sprout wheat extract.
이를 통해, 본 발명의 새싹 밀 추출물이 ROS를 억제하는 효능이 있음을 확인하였다.Through this, it was confirmed that the sprout wheat extract of the present invention has the effect of inhibiting ROS.
실시예 4. 새싹 밀 추출물의 간세포 내 GSH 함량 조절Example 4. Control of GSH Content in Hepatocytes of Sprout Wheat Extract
산화스트레스를 유도하는 TBHP에 의해 세포 내 항산화제인 글루타티온(Glutathione; GSH)의 함량 감소 및 새싹 밀 추출물을 처리하여 이의 변화를 측정하였다.The decrease in the content of intracellular antioxidant glutathione (GSH) by TBHP inducing oxidative stress and the treatment of sprout wheat extract were measured for its change.
구체적으로, 실시예 3-1과 동일하게 실험한 뒤, GSH의 함량을 측정하였다.Specifically, after the same experiment as in Example 3-1, the content of GSH was measured.
그 결과, 도 4에서 볼 수 있듯이, 세포 내 항산화제인 GSH 함량은 TBHP 처리에 의해 감소하였으나 새싹 밀 추출물 처리 시 농도의존적으로 증가하였다.As a result, as can be seen in FIG. 4 , the content of GSH, an intracellular antioxidant, was decreased by TBHP treatment, but increased in a concentration-dependent manner when treated with sprout wheat extract.
이를 통해, 본 발명의 새싹 밀 추출물이 HepG2 세포 내 GSH 함량을 증가시켜 산화스트레스에 의한 ROS 생성 억제 효능 및 세포 보호효능이 있음을 확인하였다.Through this, it was confirmed that the sprout wheat extract of the present invention increased the GSH content in HepG2 cells, thereby inhibiting ROS production by oxidative stress and protecting the cells.
실시예 5. 항산화 효소 활성 조절 효과Example 5. Antioxidant enzyme activity modulating effect
TBHP에 의해 자극되는 항산화 효소 catalase(CAT), glutathione reductase(GR) 및 glutathione peroxidase(GPx)의 새싹 밀 추출물에 의한 활성 변화를 측정하였다.The TBHP-stimulated antioxidant enzymes catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx) were measured for activity changes by sprout wheat extract.
구체적으로, 실시예 3-1과 동일하게 실험한 뒤, CAT, GR 및 GPx의 함량을 측정하였다.Specifically, after the same experiment as in Example 3-1, the contents of CAT, GR and GPx were measured.
그 결과, 도 5 내지 도 7에서 볼 수 있듯이, TBHP 처리에 의해 증가된 CAT, GR, GPx 등 항산화 효소의 활성이 새싹 밀 처리에 의해 농도 의존적으로 감소되며, 50 μg/ml 처리 시, TBHP를 처리하지 않은 대조군 수준으로 조절됨을 확인하였다.As a result, as can be seen in FIGS. 5 to 7, the activity of antioxidant enzymes such as CAT, GR, and GPx increased by TBHP treatment is reduced in a concentration-dependent manner by sprout wheat treatment, and when treated with 50 μg/ml, TBHP It was confirmed that it was adjusted to the untreated control level.
이를 통해, 본 발명의 새싹 밀 추출물이 HepG2 세포 내에서 항산화 효소의 활성을 조절하여 산화스트레스에 의한 ROS 생성 억제 효능 및 세포 보호효능이 있음을 확인하였다.Through this, it was confirmed that the sprout wheat extract of the present invention had the effect of inhibiting ROS generation by oxidative stress and protecting the cells by regulating the activity of antioxidant enzymes in HepG2 cells.
실시예 6. 새싹 밀 유래 화합물의 세포 독성 평가Example 6. Evaluation of Cytotoxicity of Sprout Wheat Derived Compounds
새싹 밀 추출물에서 추출물의 2차대사 화합물을 정제하고(도 8), 이 화합물(Isoorientin; WS1) 및 화합물(Isoschaftoside; WS2)을 농도별(0, 5, 10 및 20)로 처리하여, 화합물의 세포 독성을 측정하였다.Purifying the secondary metabolite compound of the extract from sprout wheat extract (Fig. 8), and treating this compound (Isoorientin; WS1) and compound (Isoschaftoside; WS2) by concentration (0, 5, 10 and 20), the compound Cytotoxicity was measured.
실험방법은 실시예 2와 동일하게 실험하였고, 양성 대조군으로 quercetin을 사용하였다.The experimental method was the same as in Example 2, and quercetin was used as a positive control.
그 결과, 도 9에서 볼 수 있듯이, WS1, WS2 및 querectin 모두 20 μg/mL 이하에서 세포 독성을 나타내지 않았다.As a result, as can be seen in FIG. 9, WS1, WS2 and querectin did not show cytotoxicity at 20 μg/mL or less.
이를 통해, 새싹 밀 유래 화합물이 세포에 독성을 띄지 않는 것을 확인하였다.Through this, it was confirmed that the compound derived from sprout wheat is not toxic to cells.
실시예 7. 새싹 밀 유래 화합물의 간세포 내 산화스트레스 보호 효과Example 7. Oxidative stress protective effect in hepatocytes of sprout wheat-derived compounds
인체유래 간세포인 HepG2 세포를 이용하여, 산화스트레스에 의한 세포 생존율을 측정하고, 새싹 밀 유래 화합물에 의한 산화스트레스로부터의 세포 보호 효과를 확인하였다.Cell viability by oxidative stress was measured using HepG2 cells, which are human hepatocytes, and the cytoprotective effect from oxidative stress by sprout wheat-derived compounds was confirmed.
실험은 실시예 3-1과 동일하게 실험하였고, 양성 대조군으로 quercetin을 사용하였다.The experiment was conducted in the same manner as in Example 3-1, and quercetin was used as a positive control.
그 결과, 도 10에서 볼 수 있듯이, HepG2 세포에서 산화적 스트레스를 유도하는 TBHP 처리에 의해 감소된 세포 생존율이 새싹 밀 유래 화합물 중 WS1을 처리 시 농도 의존적으로 세포 보호효과가 나타나는 반면, WS2를 처리하면 세포 보호효과가 없음을 확인하였다.As a result, as can be seen in FIG. 10, the cell viability decreased by TBHP treatment inducing oxidative stress in HepG2 cells showed a concentration-dependent cytoprotective effect upon treatment with WS1 among sprout wheat-derived compounds, whereas treatment with WS2 It was confirmed that there was no cytoprotective effect.
이를 통해, 새싹 밀 추출물 중 화합물 WS1(이소오리엔틴)이 산화스트레스로부터 간세포를 보호하는 효능이 있음을 확인하였다.Through this, it was confirmed that the compound WS1 (isoorientin) in the sprout wheat extract has the effect of protecting hepatocytes from oxidative stress.
이를 종합하면, 새싹 밀 추출물과 새싹 밀 추출물에서 유래된 화합물인 이소오리엔틴은 모두 간질환에 대한 예방 또는 치료 효과를 가지며, 또한 간 손상을 예방 또는 개선시킬 수 있음을 알 수 있다.Taken together, it can be seen that both sprout wheat extract and isoorientin, a compound derived from sprout wheat extract, have a preventive or therapeutic effect on liver disease, and can also prevent or improve liver damage.
이상의 설명으로부터, 본 발명이 속하는 기술 분야의 당업자는 본 발명이 그 기술적 사상이나 필수적 특징을 변경하지 않고서 다른 구체적인 형태로 실시될 수 있다는 것을 이해할 수 있을 것이다. 이와 관련하여, 이상에서 기술한 실시 예들은 모든 면에서 예시적인 것이며 한정적인 것이 아닌 것으로서 이해해야만 한다. 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허 청구범위의 의미 및 범위 그리고 그 등가 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.From the above description, those skilled in the art to which the present invention pertains will understand that the present invention may be embodied in other specific forms without changing the technical spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention, rather than the above detailed description, all changes or modifications derived from the meaning and scope of the claims described below and their equivalents.
Claims (12)
A food composition for preventing or improving non-alcoholic liver function damage comprising a sprout wheat extract or a compound derived therefrom as an active ingredient.
The food composition according to claim 1, wherein the compound is isoorientin (isoorientin).
The food composition of claim 1, wherein the liver function impairment is due to oxidative stress.
The food composition according to claim 1, wherein the extract is obtained by extracting sprout wheat with a solvent selected from the group consisting of water, lower alcohols having 1 to 4 carbon atoms, and mixed solvents thereof.
The food composition according to claim 1, wherein the composition protects hepatocytes from oxidative stress.
The food composition of claim 5, wherein the protection is achieved by inhibiting reactive oxygen species (ROS) production, increasing glutathione, and regulating the activity of antioxidant enzymes.
The food composition of claim 6, wherein the antioxidant enzyme is catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx).
A feed composition for preventing or improving non-alcoholic liver function damage, comprising a sprout wheat extract or a compound derived therefrom as an active ingredient.
A pharmaceutical composition for preventing or treating liver disease, comprising a wheat sprout extract or a compound derived therefrom as an active ingredient.
The pharmaceutical composition according to claim 9, wherein the liver disease is caused by impaired liver function by oxidative stress.
The pharmaceutical composition of claim 10, wherein the liver disease is non-alcoholic liver injury.
The pharmaceutical composition of claim 9, wherein the composition further comprises a pharmaceutically acceptable carrier, excipient or diluent.
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| KR1020220114174A KR20220128971A (en) | 2019-12-12 | 2022-09-08 | Composition comprising sprout wheat-derived for improvement of hepatopathy |
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| CN114869901A (en) * | 2022-05-06 | 2022-08-09 | 安徽医科大学 | Application of verbena glycoside in medicine for treating alcoholic liver injury |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114869901A (en) * | 2022-05-06 | 2022-08-09 | 安徽医科大学 | Application of verbena glycoside in medicine for treating alcoholic liver injury |
| CN114869901B (en) * | 2022-05-06 | 2023-10-27 | 安徽医科大学 | Application of verbena glycoside in medicine for treating alcoholic liver injury |
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