KR20150051159A - Composition for preventing or treating thyroid disorders comprising phytolacca esculenta houttuyn extracts or fraction thereof - Google Patents
Composition for preventing or treating thyroid disorders comprising phytolacca esculenta houttuyn extracts or fraction thereof Download PDFInfo
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- KR20150051159A KR20150051159A KR1020140148429A KR20140148429A KR20150051159A KR 20150051159 A KR20150051159 A KR 20150051159A KR 1020140148429 A KR1020140148429 A KR 1020140148429A KR 20140148429 A KR20140148429 A KR 20140148429A KR 20150051159 A KR20150051159 A KR 20150051159A
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Abstract
Description
본 발명은 상륙(Phytolacca esculenta Houttuyn) 추출물 또는 이의 분획물을 함유하는 갑상선 질환의 예방, 개선 또는 치료용 조성물에 관한 것이다. 구체적으로, 본 발명은 상륙 추출물 또는 이의 분획물을 유효 성분으로 함유하는 갑상선 질환의 예방 또는 치료용 약학 조성물, 또는 갑상선 질환의 예방 또는 개선용 식품 조성물에 관한 것이며, 상기 조성물을 이용하여 갑상선 질환을 예방, 개선 또는 치료하는 방법에 관한 것이다.
The present invention is based on the finding that landing ( Phytolacca esculenta Houttuyn) extract or a fraction thereof, to a composition for preventing, ameliorating or treating thyroid disease. Specifically, the present invention relates to a pharmaceutical composition for prevention or treatment of thyroid disease, which contains Land Root Extract or its fraction as an active ingredient, or a food composition for preventing or ameliorating thyroid disease, , ≪ / RTI >
갑상선은 요오드를 원료로 갑상선 호르몬을 만들어 혈류로 분비하는 내분비 기관이다. 갑상선 호르몬은 우리 몸의 신진대사의 속도를 조절하는 역할을 한다. 이러한 갑상선 호르몬 중에서 가장 중요한 것은 T4(요오드 원자4개)라고 불리는 티록신(thyroxine)과 T3(요오드 원자 3개)라고 불리는 트리요오드티로닌(triiodothyronine)이다. 이들 호르몬은 신체의 성장 발육과 인체에 필요한 당질, 단백질, 지방질 등의 생성 등, 몸이 스스로를 유지해나가는 각종 물질대사 과정을 조절한다. 또한 대사 과정을 촉진시켜 에너지를 만드는 역할을 하여 열을 발생시킴으로써 체온 조절에도 관여하여 모든 기관이 제 기능을 적절히 유지하도록 한다.
Thyroid is an endocrine organ that produces iodine as a raw material and secretes thyroid hormone into the bloodstream. Thyroid hormones control the rate of metabolism in our bodies. The most important of these thyroid hormones is thyroxine called T4 (four iodine atoms) and triiodothyronine called T3 (three iodine atoms). These hormones regulate various metabolic processes that the body maintains itself, such as the growth and development of the body and the production of carbohydrates, proteins and fats necessary for the human body. It also promotes metabolic processes to generate energy, which generates heat, which is involved in body temperature control, so that all organs can function properly.
최근 갑상선 질환 환자의 수가 급증하고 있는데, 갑상선 질환의 주된 원인 중 하나가 갑상선 호르몬의 분비 조절의 이상에 있다.Recently, the number of patients with thyroid disease is rapidly increasing. One of the main causes of thyroid disease is abnormality of thyroid hormone secretion control.
체내에서 갑상선 호르몬의 분비를 위해서는 뇌하수체에서 갑상선 자극 호르몬(Thyroid Stimulating Hormone; TSH)이 분비되어야 한다. 이는 피드백 작용에 의해서 조절되는데, 구체적으로 TSH의 분비가 억제된 상태이면 갑상선에서 호르몬 분비에 대한 자극이 감소되어 갑상선 호르몬이 부족하게 되며, 이에 뇌하수체에서 TSH의 분비를 촉진하게 되어 갑상선 호르몬 분비가 증가되도록 한다. 반대로 TSH의 분비가 촉진된 상태이면 갑상선 호르몬이 과잉 분비되어, 이에 뇌하수체에서 TSH의 분비를 억제하게 되어 갑상선 호르몬 분비가 억제되도록 한다.For the secretion of thyroid hormones in the body, thyroid stimulating hormone (TSH) should be secreted from the pituitary gland. This is regulated by feedback. Specifically, when the secretion of TSH is suppressed, the stimulation of hormone secretion is reduced in the thyroid gland and the thyroid hormone becomes deficient. Thus, the secretion of TSH from the pituitary gland is promoted and the thyroid hormone secretion is increased . Conversely, when TSH secretion is promoted, excess thyroid hormone is secreted, which inhibits the secretion of TSH from the pituitary gland and suppresses thyroid hormone secretion.
갑상선 호르몬 분비 조절 이상에 따른 질환으로는 대표적으로 갑상선 기능 항진증(hyperthyroidism) 및 갑상선 기능 저하증(hypothyroidism)이 있다. 갑상선 기능 항진증은 갑상선 호르몬이 혈액내로 과다 방출되어 일어나는 증상을 가진 모든 질환을 총칭한다. 갑상선 기능 항진증의 주요 특징은 갑상선이 증대되고 기초 대사율이 증가하며 자율 신경계통의 실조를 야기하는 것인데, 최근 통계에 따르면 여성이 남성에 비해 4:1의 비율로 많이 발생하며, 그 중에서도 20대 내지 50대에서 많이 발생한다.Hyperthyroidism and hypothyroidism are the most common disorders associated with abnormal thyroid hormone secretion. Hyperthyroidism refers to all diseases with symptoms that occur when thyroid hormones are overdosed into the blood. The main features of hyperthyroidism are thyroid hyperplasia, increased basal metabolic rate, and causing autonomic nervous system failure. According to recent statistics, women are more likely to develop at a ratio of 4: 1 compared to men, It occurs a lot in 50s.
갑상선 기능 저하증이란 갑상선 호르몬의 생성 부족으로 인한 모든 질환을 총칭한다. 갑상선 기능 저하증은 일반적으로 매우 서서히 진행하는 질환이므로 인식하기 어려운 경우가 많으며, 그 증상이 질환의 진행 정도에 따라 매우 다양하나, 일반적으로 우리 몸의 대사 상태가 감소되어 쉽게 피로해지고 무기력해지며 의욕이 상실되고, 추위에 민감해지며 체중이 증가하고 기억력 감퇴, 식욕 감퇴, 근육통, 관절통, 발한 감소 등의 증상을 야기한다.
Hypothyroidism is a general term for all diseases caused by lack of production of thyroid hormones. Hypothyroidism is usually a very slowly progressing disease, and it is often difficult to recognize. The symptoms vary widely depending on the progression of the disease, but generally the metabolic state of our body is reduced, making it easily fatigued and helpless. Lose weight, become sensitive to cold, increase weight, and cause symptoms such as decreased memory, loss of appetite, muscle aches, arthralgia, and decreased sweating.
이러한 갑상선 기능 항진증 및 저하증을 포함하는 갑상선 질환은 갑상선 호르몬과 관련된 것이 대부분이며, 따라서 현재 갑상선 질환을 치료하기 위해서는 호르몬 등을 외부로부터 체내로 공급해야 하는 등 지속적인 약물 치료에 의존해야 하는 경우가 대부분이어서, 급성 심근경색이나 불안정성 협심증 등의 허혈성 심질환 유발의 위험성, 환자의 체내 갑상선 호르몬 요구량의 변화 등 여러 문제점이 나타나는 것으로 보고되고 있다. 따라서 부작용이 적고 적용이 보다 용이한 갑상선 질환 치료제의 개발이 요구되는 실정이다.
Thyroid diseases including hyperthyroidism and hypothyroidism are mostly related to thyroid hormones. Therefore, in order to treat thyroid diseases at present, most of them have to rely on continuous medication such as supplying hormones to the body from the outside , The risk of ischemic heart disease induction such as acute myocardial infarction or unstable angina, and changes in the body's thyroid hormone requirement. Therefore, it is required to develop a therapeutic agent for thyroid disease which has fewer side effects and is easier to apply.
이러한 배경하에, 본 발명자들은 갑상선 질환을 치료할 수 있는 물질에 대한 연구를 수행하였으며, 이에 상륙(Phytolacca esculenta Houttuyn) 추출물이 갑상선 호르몬의 분비를 조절하는 효능이 있음을 규명하고, 이를 활용한 갑상선 질환의 예방, 개선 또는 치료용 조성물을 완성하기에 이르렀다.
Under this background, the present inventors have carried out research on the matter to treat thyroid disorders, and thus landing (Phytolacca esculenta Houttuyn extract has been found to be effective in regulating the secretion of thyroid hormone and has been completed to prepare a composition for preventing, ameliorating or treating thyroid disease using the extract.
본 발명은 상륙 추출물 또는 이의 분획물을 유효 성분으로 함유하는 조성물을 이용하여 갑상선 질환을 예방, 개선 또는 치료하는 것을 목적으로 한다.The present invention aims to prevent, ameliorate, or treat thyroid disease by using a composition containing Land Root Extract or its fraction as an active ingredient.
구체적으로, 본 발명의 하나의 목적은 상륙 추출물 또는 이의 분획물을 유효 성분으로 함유하는 갑상선 질환의 예방 또는 치료용 조성물, 또는 갑상선 질환의 예방 또는 개선용 식품 조성물을 제공하는 것이다.Specifically, one object of the present invention is to provide a composition for preventing or treating a thyroid disease or a food composition for preventing or ameliorating a thyroid disease, which comprises a landing extract or a fraction thereof as an active ingredient.
본 발명의 다른 목적은 상기 조성물을 개체에 투여하는 것을 포함하는 갑상선 질환의 예방, 개선 또는 치료 방법을 제공하는 것이다.
It is another object of the present invention to provide a method for preventing, ameliorating or treating thyroid disease, which comprises administering the composition to a subject.
본 발명은 상기의 과제를 해결하기 위한 하나의 구현예로서, 상륙 추출물 또는 이의 분획물을 유효 성분으로 함유하는 갑상선 질환의 예방 또는 치료용 조성물을 제공한다.The present invention provides, as one embodiment for solving the above problems, a composition for preventing or treating thyroid disease, which comprises an immersion extract or a fraction thereof as an active ingredient.
본 발명의 상륙(商陸; Phytolacca esculenta Houttuyn)은 자리공과(Phytolaccaceae)에 속하는 다년생 초본으로 자리공이라고도 불리며, 전초(全草)가 약으로 사용될 수 있는 식물이다. 열대 및 아열대 특히 아메리카 등지에 약 35종이 분포하고 있으며 우리나라에는 1속 2종이 분포되어 있다. 한국과 일본에서는 Phytolacca esculenta 및 그 동속 식물을 같은 기원 식물로 사용하고 있으며, 중국에서는 Phytolacca acinosa 및 Phytolacca americana 식물이 같은 기원 식물로 사용되고 있다. 상륙으로부터 분리된 화합물로는 페놀계열의 갈산(gallic acid), 프로토카테규산(protocatechuic acid), 클로로제닉산(clorogenic acid), 카페인산(caffeic acid), m-하이드록시벤조산(m-hydroxybenzoic acid), p-쿠마릭산(coumaric acid) 및 시나믹산(cinnamic acid) 등이 있으며, 사포닌계열의 에스큘렌토사이드(esculentoside) A, E, F, H 등이 알려져 있으며, 이외에도 트리테르펜(triterpene) 성분도 알려져 있다.
The landing (commercial; Phytolacca esculenta Houttuyn) is a perennial herb that belongs to the phytolaccaceae and is also known as a perennial herb, and can be used as a herbal medicine. About 35 species are distributed in the tropical and subtropical regions, especially in the Americas, and 2 species in the first category are distributed in Korea. In Korea and Japan, Phytolacca esculenta and its subordinate plants are used as the same origin plant, and in China Phytolacca acinosa and Phytolacca Americana plants are used as the same origin plants. Compounds isolated from landings include phenolic compounds such as gallic acid, protocatechuic acid, clorogenic acid, caffeic acid, m-hydroxybenzoic acid, coumaric acid and cinnamic acid. Saponin esculentosides A, E, F and H are known. In addition, triterpene components are also known. have.
본 발명에서 사용되는 용어, "추출물"은 상륙의 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 추출물 또는 분획물은 바람직하게는 추출 후 건조 분말 형태로 제조되어 사용될 수 있다.The term "extract" used in the present invention means an extract obtained by an extraction treatment of landing, a diluted solution or a concentrate of the extract, a dried product obtained by drying the extract, a controlled preparation or a purified product of the extract, Extracts themselves and extracts of all formulations which can be formed using extracts. The extract or fraction of the present invention can be prepared and used in the form of a dry powder after extraction.
본 발명의 상기 상륙 추출물은, 상기 상륙의 천연, 잡종 또는 변종 식물의 다양한 기관으로부터 추출될 수 있고, 예를 들어 상륙의 뿌리, 지상부, 줄기, 잎, 꽃, 열매의 몸통, 열매의 껍질, 종자뿐만 아니라 식물 조직 배양물로부터도 추출이 가능하다.The landing extract of the present invention can be extracted from various organs of the natural, hybrid, or variant plants of the above landing, and can be extracted from various organs such as roots, land parts, stems, leaves, flowers, fruit bodies, It can also be extracted from plant tissue culture.
본 발명의 상기 상륙 추출물은 바람직하게는 상륙의 뿌리, 줄기 또는 잎의 추출물일 수 있다.The landing extract of the present invention may preferably be an extract of root, stem or leaf of landing.
본 발명의 상기 상륙 추출물에 있어서, 상기 상륙을 추출하는 방법은 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 추출할 수 있다. 상기 추출 방법의 비제한적인 예로는, 열수 추출법, 초음파 추출법, 여과법, 환류 추출법 등을 들 수 있으며, 이들은 단독으로 수행되거나 2 종 이상의 방법을 병용하여 수행될 수 있다.In the landing extract of the present invention, the method for extracting the landing is not particularly limited, and may be extracted according to a method commonly used in the art. Non-limiting examples of the extraction method include hydrothermal extraction, ultrasonic extraction, filtration, and reflux extraction. These may be performed alone or in combination with two or more methods.
본 발명에서 상기 상륙을 추출하는 데에 사용되는 추출 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 추출 용매의 비제한적인 예로는 물, 알코올 또는 이들의 혼합 용매 등을 들 수 있으며, 알코올을 용매로 사용하는 경우에는 보다 바람직하게는 C1 내지 C4의 알코올을 사용할 수 있다. 본 발명의 상륙 추출물은 바람직하게는 물 또는 에탄올 추출물일 수 있다.
In the present invention, the kind of the extraction solvent used for extracting the landing is not particularly limited, and any solvent known in the art can be used. Nonlimiting examples of the extraction solvent include water, alcohols or mixed solvents thereof. When alcohol is used as a solvent, C 1 to C 4 alcohols may be more preferably used. The landing extract of the present invention may preferably be water or an ethanol extract.
본 발명에서 사용되는 용어, "분획물"은 여러 다양한 구성 성분들을 포함하는 혼합물로부터 특정 성분 또는 특정 성분 그룹을 분리하기 위하여 분획을 수행하여 얻어진 결과물을 의미한다.The term "fraction " as used herein means a product obtained by performing fractionation to separate a specific component or a specific component group from a mixture containing various components.
본 발명에서 상기 분획물을 얻는 분획 방법은 특별히 제한되지 아니하며, 당해 기술 분야에서 통상적으로 사용하는 방법에 따라 수행될 수 있다. 상기 분획 방법의 비제한적인 예로는, 상륙을 추출하여 얻은 추출물에 소정의 용매를 처리하여 상기 추출물로부터 분획물을 얻는 방법을 들 수 있다.The fractionation method for obtaining the fraction in the present invention is not particularly limited and may be carried out according to a method commonly used in the art. As a non-limiting example of the above-mentioned fractionation method, there can be enumerated a method of obtaining a fraction from the extract by treating a predetermined solvent with an extract obtained by extracting landfall.
본 발명에서 상기 분획물을 얻는 데에 사용되는 분획 용매의 종류는 특별히 제한되지 아니하며, 당해 기술 분야에서 공지된 임의의 용매를 사용할 수 있다. 상기 분획 용매의 비제한적인 예로는 물, 알코올 등의 극성 용매; 헥산(Hexan), 에틸 아세테이트(Ethyl acetate), 클로로포름(Chloroform), 디클로로메탄(Dichloromethane) 등의 비극성 용매 등을 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상 혼합하여 사용될 수 있다. 상기 분획 용매 중 알코올을 사용하는 경우에는 바람직하게는 C1 내지 C4의 알코올을 사용할 수 있다.
The kind of the fraction solvent used for obtaining the fraction in the present invention is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fraction solvent include polar solvents such as water and alcohol; And non-polar solvents such as hexane, ethyl acetate, chloroform, and dichloromethane. These may be used alone or in combination of two or more. When alcohols are used in the fraction solvent, C 1 to C 4 alcohols can be preferably used.
본 발명의 상기 상륙 추출물 또는 이의 분획물은 갑상선 호르몬, 특히 티록신의 분비를 조절하는 효능을 나타내며, 이는 본 발명에서 최초로 규명된 것이다. 구체적으로 본 발명의 상륙 추출물 또는 이의 분획물을 함유하는 조성물은 갑상선 자극 호르몬(TSH)이 과잉된 조건에서는 티록신의 분비를 감소시키고, TSH가 부족한 조건에서는 티록신의 분비를 증가시켜 티록신의 분비를 조절하는 효능을 나타낸다.The landing extract or fraction thereof of the present invention exhibits an effect of regulating the secretion of thyroid hormone, in particular thyroxine, which was first identified in the present invention. Specifically, the composition containing the landing extract or its fractions of the present invention reduces the secretion of thyroxine under the condition that thyroid stimulating hormone (TSH) is excessive and regulates thyroxine secretion by increasing the secretion of thyroxine under the condition of lacking TSH Show efficacy.
본 발명의 구체적인 일 실시예에 따르면, TSH가 포함된 배지(6H 배지) 및 TSH가 포함되지 않은 배지(5H 배지) 각각에 본 발명의 상륙 추출물을 제공하고, 상기 각 배지에서 배양된 랫트의 갑상선 세포주 FRTL-5 세포에서의 티록신 분비 변화를 측정한 결과, 상륙 추출물 무처리 대조군과 비교하여, 6H 배지에서는 티록신의 분비가 감소되고, 5H 배지에서는 반대로 티록신의 분비가 증가되는 결과를 나타내어, 상륙 추출물이 티록신의 분비를 조절하는 효능이 있음을 확인할 수 있었다(실시예 2 및 3).
According to a specific embodiment of the present invention, the landing extract of the present invention is provided to each of the medium containing TSH (6H medium) and the medium not containing TSH (5H medium), and the thyroid gland As a result of measuring the change of thyroxine secretion in cell line FRTL-5 cells, the secretion of thyroxine was decreased in 6H medium and the secretion of thyroxine was reversed in 5H medium compared to the control without landing extract, And it was confirmed that there was an effect of controlling the secretion of thyroxine (Examples 2 and 3).
본 발명의 구체적인 일 실시예에 따르면, TSH가 포함되지 않은 배지(5H 배지)에 상륙 추출물을 처리한 경우, FRTL-5 세포의 증식을 유도함을 확인하였으며, 이러한 효과는 estradiol과 시너지 효과를 가짐을 확인하였다(실시예 4).
According to a specific embodiment of the present invention, it has been confirmed that when the landing extract is treated with a medium containing no TSH (5H medium), FRTL-5 cell proliferation is induced, and this effect has a synergistic effect with estradiol (Example 4).
또한, 본 발명의 상기 조성물은 갑상선 질환의 예방, 개선 또는 치료 효과를 충분히 나타내는 농도 범위에서 세포 독성을 나타내지 않으며, 천연물로부터 유래된 것이어서 체내에 심각한 자극을 가한다거나 유해한 작용을 유발함이 없이 안정하게 사용될 수 있는 이점이 있다.
In addition, the composition of the present invention does not exhibit cytotoxicity in a concentration range sufficiently exhibiting the preventive, ameliorating or therapeutic effect of thyroid disease, and is derived from a natural product, so that the composition stably stimulates the body without causing serious irritation or causing harmful action There is an advantage that can be used.
본 발명에서 사용되는 용어, "갑상선 질환"은 갑상선에 야기된 모든 질환 및 갑상선의 기능 이상 증상을 나타내는 모든 질환을 의미한다. 상기 갑상선 질환의 비제한적인 예로 갑상선 기능 항진증, 갑상선 기능 저하증, 갑상선염, 갑상선 결절, 갑상선암 등을 들 수 있다. 본 발명의 상기 갑상선 질환은 특별히 제한되지 아니하나, 본 발명의 목적상, 바람직하게는 갑상선 기능 항진증 및/또는 갑상선 기능 저하증을 의미할 수 있다.As used herein, the term "thyroid disease" refers to any disease that causes thyroid disease and dysfunctional symptoms of the thyroid gland. Non-limiting examples of the thyroid disease include hyperthyroidism, hypothyroidism, thyroiditis, thyroid nodule, thyroid cancer and the like. The disease of the thyroid according to the present invention is not particularly limited, but it may preferably mean hyperthyroidism and / or hypothyroidism for the purpose of the present invention.
상기 갑상선 기능 항진증(hyperthyroidism)이란 갑상선 호르몬의 과잉 생성으로 인한 모든 질환을 총칭하며, 갑상선 기능 저하증(hypothyroidism)이란 갑상선 호르몬의 생성 부족으로 인한 모든 질환을 총칭한다.
The hyperthyroidism refers to all diseases caused by excessive production of thyroid hormone. Hypothyroidism refers to all diseases caused by lack of production of thyroid hormone.
본 발명에서 사용되는 용어, "예방"이란, 본 발명의 상기 조성물을 개체에 투여하여 갑상선 질환의 발병을 억제시키거나 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" means any action that inhibits or delays the onset of thyroid disease by administering the composition of the invention to a subject.
본 발명에서 사용되는 용어, "치료"란, 본 발명의 상기 조성물을 개체에 투여하여 갑상선 질환의 증세가 호전되도록 하거나 이롭게 되도록 하는 모든 행위를 의미한다.
The term "treatment" as used in the present invention means all the actions that cause the symptom of thyroid disease to be improved or benefited by administering the composition of the present invention to an individual.
본 발명의 상기 약학적 조성물에 있어서, 상기 상륙 추출물 또는 이의 분획물은 상기 약학적 조성물의 전체의 중량을 기준으로 바람직하게는 1 중량% 내지 99.99 중량%로 함유될 수 있고, 보다 바람직하게는 10 중량% 내지 99.99 중량%로 함유될 수 있으며, 더욱 바람직하게는 50 중량% 내지 99.99 중량%로 함유될 수 있다. 상기 범위내에서, 상기 상륙 추출물 또는 이의 분획물에 따른 갑상선 호르몬 분비 조절 효과를 비롯한 갑상선 질환 치료 효과가 충분히 발휘되어 본 발명의 목적을 달성하기에 보다 적합해지는 이점이 있다.
In the pharmaceutical composition of the present invention, the abovementioned landing extract or its fraction may be contained in an amount of preferably 1% by weight to 99.99% by weight, more preferably 10% by weight % To 99.99% by weight, and more preferably 50% by weight to 99.99% by weight. Within the above range, the therapeutic effect of the thyroid disease including the thyroid hormone secretion-controlling effect according to the landing extract or its fraction is sufficiently exhibited, which is advantageous to achieve the object of the present invention.
본 발명의 상기 약학적 조성물은, 상기 상륙 추출물 또는 이의 분획물을 유효 성분으로 함유하는 것에 더하여, 약학적으로 허용 가능한 담체를 추가로 포함할 수 있다.The pharmaceutical composition of the present invention may further comprise a pharmaceutically acceptable carrier in addition to the above landing extract or a fraction thereof as an active ingredient.
본 발명에서, 상기 "약학적으로 허용 가능"하다는 것은, 이를 투여 시 생물체를 자극하지 않으면서, 투여되는 화합물의 생물학적 활성 및 특성을 저해하지 않는, 약학 분야에서 통상적으로 사용되는 것을 의미한다.In the present invention, the above-mentioned "pharmaceutically acceptable" means that it is commonly used in the pharmaceutical field, which does not disturb the biological activity and properties of the compound administered, without irritating the organism upon administration thereof.
본 발명의 상기 약학적 조성물은, 상기 담체와 함께 제제화되어, 식품, 의약품, 사료 첨가제, 음용수 첨가제 등으로 활용될 수 있다.The pharmaceutical composition of the present invention may be formulated together with the carrier to be used as food, medicines, feed additives, drinking water additives, and the like.
본 발명에서, 상기 담체의 종류는 특별히 제한되지 아니하며 당해 기술 분야에서 통상적으로 사용되는 담체라면 어느 것이든 사용할 수 있다. 상기 담체의 비제한적인 예로는, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사 용액, 락토오스, 덱스트로오스, 수크로오스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 말토 덱스트린, 글리세롤, 에탄올 등을 들 수 있다. 이들은 단독으로 사용되거나 2 종 이상을 혼합하여 사용될 수 있다.In the present invention, the type of the carrier is not particularly limited and any carrier conventionally used in the art can be used. Examples of the carrier include, but are not limited to, saline, sterilized water, Ringer's solution, buffered saline, albumin injection solution, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, maltodextrin, glycerol, . These may be used alone or in combination of two or more.
또한, 본 발명의 상기 약학적 조성물은 필요한 경우, 부형제, 희석제, 항산화제, 완충액 또는 정균제 등 기타 약학적으로 허용 가능한 첨가제 들을 첨가하여 사용할 수 있으며, 충진제, 증량제, 습윤제, 붕해제, 분산제, 계면 활성제, 결합제 또는 윤활제 등을 부가적으로 첨가하여 사용할 수 있다.
In addition, the pharmaceutical composition of the present invention may be supplemented with other pharmaceutically acceptable additives such as excipients, diluents, antioxidants, buffers or bacteriostats, if necessary, and may be used as fillers, extenders, wetting agents, disintegrants, An activator, a binder, a lubricant, or the like may be additionally used.
본 발명의 상기 약학적 조성물은 경구 투여 또는 비경구 투여를 위한 적합한 다양한 제형으로 제제화되어 사용될 수 있다.The pharmaceutical composition of the present invention may be formulated into various formulations suitable for oral administration or parenteral administration.
상기 경구 투여용 제제의 비제한적인 예로는, 트로키제(troches), 로젠지(lozenge), 정제, 수용성 현탁액, 유성 현탁액, 조제 분말, 과립, 에멀젼, 하드 캡슐, 소프트 캡슐, 시럽 또는 엘릭시르제 등을 들 수 있다.Non-limiting examples of such oral dosage forms include troches, lozenges, tablets, aqueous suspensions, oily suspensions, prepared powders, granules, emulsions, hard capsules, soft capsules, syrups or elixirs .
본 발명의 상기 약학적 조성물을 경구 투여용으로 제제화하기 위하여, 락토오스, 사카로오스, 솔비톨, 만니톨, 전분, 아밀로펙틴(Amylopectin), 셀룰로오스(Cellulose) 또는 젤라틴(Gelatin) 등과 같은 결합제; 디칼슘 포스페이트(Dicalcium phosphate) 등과 같은 부형제; 옥수수 전분 또는 고구마 전분 등과 같은 붕괴제; 스테아르산 마그네슘(Magnesium stearate), 스테아르산 칼슘(Calcium stearate), 스테아릴 푸마르산 나트륨(Sodium stearyl fumarate) 또는 폴리에틸렌 글리콜 왁스(Polyethylene glycol wax) 등과 같은 윤활유 등을 사용할 수 있으며, 감미제, 방향제, 시럽제 등도 사용할 수 있다.In order to formulate the pharmaceutical composition of the present invention for oral administration, a binder such as lactose, saccharose, sorbitol, mannitol, starch, amylopectin, cellulose, or gelatin; Excipients such as dicalcium phosphate and the like; Disintegrating agents such as corn starch or sweet potato starch; Lubricants such as magnesium stearate, calcium stearate, sodium stearyl fumarate or polyethylene glycol wax may be used, and sweeteners, fragrances, syrups and the like may also be used. .
나아가 캡슐제의 경우에는 상기 언급한 물질 외에도 지방유와 같은 액체 담체 등을 추가로 사용할 수 있다.
Furthermore, in the case of capsules, in addition to the above-mentioned substances, liquid carriers such as fatty oils can be further used.
상기 비경구용 제제의 비제한적인 예로는, 주사액, 좌제, 호흡기 흡입용 분말, 스프레이용 에어로졸제, 연고, 도포용 파우더, 오일, 크림 등을 들 수 있다.Non-limiting examples of the parenteral preparation include injections, suppositories, respiratory inhalation powders, aerosol preparations for spraying, ointments, powder for application, oils, creams and the like.
본 발명의 상기 약학적 조성물을 비경구 투여용으로 제제화하기 위하여, 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결 건조 제제, 외용제 등을 사용할 수 있으며, 상기 비수성용제, 현탁제로는 프로필렌글리콜, 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다.In order to formulate the pharmaceutical composition of the present invention for parenteral administration, a sterilized aqueous solution, a non-aqueous solvent, a suspending agent, an emulsion, a lyophilized preparation, an external agent, and the like may be used. Examples of the non-aqueous solution and the suspension include propylene glycol, Polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like.
또한, 보다 구체적으로 본 발명의 상기 약학적 조성물을 주사액으로 제제화하는 경우, 본 발명의 상기 조성물을 안정제 또는 완충제와 함께 물에서 혼합하여 용액 또는 현탁액으로 제조하고 이를 앰플(ampoule) 또는 바이알(vial)의 단위 투여용으로 제제화할 수 있다. 또한, 본 발명의 상기 약학적 조성물을 에어로졸제로 제제화하는 경우, 수분산된 농축물 또는 습윤 분말이 분산되도록 추진제 등이 첨가제와 함께 배합할 수 있다.More specifically, when the pharmaceutical composition of the present invention is formulated into an injection, the composition of the present invention is mixed with water or a stabilizer or a buffer in water to prepare a solution or suspension, which is then mixed with an ampoule or a vial, And the like. When the pharmaceutical composition of the present invention is formulated into an aerosol formulation, a propellant or the like may be added together with the additive such that the water-dispersed concentrate or the wet powder is dispersed.
또한, 본 발명의 상기 약학적 조성물을 연고, 크림 등으로 제제화하는 경우에는, 동물성 유, 식물성 유, 왁스, 파라핀, 전분, 트라칸트, 셀룰로오스 유도체, 폴리에틸렌 글리콜, 실리콘, 벤토나이트, 실리카, 탈크, 산화 아연 등을 담체로 사용하여 제제화할 수 있다.
When the pharmaceutical composition of the present invention is formulated into an ointment, cream, or the like, it may be an animal oil, a vegetable oil, a wax, a paraffin, a starch, a tracer, a cellulose derivative, a polyethylene glycol, a silicone, a bentonite, Zinc or the like as a carrier.
본 발명의 상기 약학적 조성물의 약학적 유효량, 유효 투여량은 상기 약학적 조성물의 제제화 방법, 투여 방식, 투여 시간 및/또는 투여 경로 등에 의해 다양해질 수 있으며, 상기 약학적 조성물의 투여로 달성하고자 하는 반응의 종류와 정도, 투여 대상이 되는 개체의 종류, 연령, 체중, 일반적인 건강 상태, 질병의 증세나 정도, 성별, 식이, 배설, 해당 개체에 동시 또는 이시에 함께 사용되는 약물 기타 조성물의 성분 등을 비롯한 여러 인자 및 의약 분야에서 잘 알려진 유사 인자에 따라 다양해질 수 있으며, 당해 기술 분야에서 통상의 지식을 가진 자는 목적하는 치료에 효과적인 투여량을 용이하게 결정하고 처방할 수 있다.The pharmaceutically effective amount and the effective dose of the pharmaceutical composition of the present invention may be varied depending on the formulation method, administration method, administration time and / or route of administration of the pharmaceutical composition. The type and severity of the response, the type of subject being treated, the age, body weight, general health status, severity or severity of the disease, sex, diet, excretion, drugs used simultaneously or simultaneously with the subject, , And the like, and those skilled in the art will readily determine and prescribe dosages that are effective for the desired treatment.
본 발명의 상기 약학적 조성물의 보다 바람직한 효과를 위한 투여량은, 바람직하게는 1일 1 mg/kg 내지 1,000 mg/kg, 보다 바람직하게는 10 mg/kg 내지 500 mg/kg일 수 있다. 본 발명의 상기 약학적 조성물의 투여는 하루에 1 회 투여될 수 있고, 수회에 나누어 투여될 수 있다. 따라서 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.
The dose for the more preferable effect of the pharmaceutical composition of the present invention may be preferably 1 mg / kg to 1,000 mg / kg per day, more preferably 10 mg / kg to 500 mg / kg per day. Administration of the pharmaceutical composition of the present invention can be administered once a day, or divided into several doses. Accordingly, the dosage is not limited in any way to the scope of the present invention.
본 발명의 상기 약학적 조성물의 투여 경로 및 투여 방식은 각각 독립적일 수 있으며, 그 방식에 있어 특별히 제한되지 아니하며, 목적하는 해당 부위에 상기 약학적 조성물이 도달할 수 있는 한 임의의 투여 경로 및 투여 방식에 따를 수 있다. 상기 약학적 조성물은 경구 투여 또는 비경구 투여 방식으로 투여할 수 있다.The route of administration and the mode of administration of the pharmaceutical composition of the present invention may be independent of each other and are not particularly limited in the method and may be arbitrarily administered and administered as long as the pharmaceutical composition can reach the desired site It is possible to follow the method. The pharmaceutical composition may be administered orally or parenterally.
상기 비경구 투여하는 방법으로는, 예를 들어 정맥 내 투여, 복강 내 투여, 근육 내 투여, 경피 투여 또는 피하 투여 등을 이용할 수 있으며, 상기 조성물을 질환 부위에 도포하거나 분무, 흡입하는 방법 또한 이용할 수 있으나 이들에 제한되지 아니한다.The parenteral administration may be, for example, intravenous administration, intraperitoneal administration, intramuscular administration, transdermal administration or subcutaneous administration, and a method of applying, spraying or inhalation the composition to a diseased site may also be used But are not limited to.
본 발명의 상기 약학적 조성물은 바람직하게는 경구 투여 또는 주사 투여될 수 있다.
The pharmaceutical composition of the present invention can preferably be administered orally or by injection.
본 발명의 또 다른 일 구현예에 따르면, 본 발명의 상기 약학적 조성물을 개체에 투여하여 갑상선 질환을 예방 또는 치료하는 방법을 제공한다.According to another embodiment of the present invention, there is provided a method for preventing or treating thyroid disease by administering the pharmaceutical composition of the present invention to an individual.
본 발명에서 사용되는 용어, "개체"는 쥐, 가축, 인간 등을 포함하는 포유 동물을 비롯한 모든 동물을 의미한다.The term "individual" as used herein refers to all animals, including mammals including rats, livestock, humans, and the like.
본 발명의 상기 갑상선 질환의 예방 또는 치료 방법에서, 상기 약학적 조성물의 투여량, 투여 경로, 투여 방식 등은 본 발명의 상기 약학적 조성물과 관련하여 상기에서 설명한 바와 동일하다.In the method for the prevention or treatment of thyroid disease according to the present invention, the dose, route of administration, mode of administration, etc. of the pharmaceutical composition are the same as those described above in connection with the pharmaceutical composition of the present invention.
본 발명의 또 다른 일 구현예에 따르면, 상륙 추출물 또는 이의 분획물을 유효 성분으로 함유하는 갑상선 질환의 예방 또는 개선용 식품 조성물을 제공한다.According to another embodiment of the present invention, there is provided a food composition for preventing or ameliorating thyroid disease, which comprises an ashram extract or a fraction thereof as an active ingredient.
본 발명의 상기 식품 조성물은 특별히 제한되지 아니하며, 건강 기능 식품 조성물을 포함한다.The food composition of the present invention is not particularly limited and includes a health functional food composition.
본 발명의 상기 건강 기능 식품 조성물을 식품 첨가물로 사용할 경우, 상기 조성물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다.When the health functional food composition of the present invention is used as a food additive, the composition may be added as it is or may be used together with other food or food ingredients, and may be appropriately used according to a conventional method.
상기 식품의 종류는 특별히 제한되지 아니하며, 통상적인 의미에서의 식품을 모두 포함한다. 상기 물질을 첨가할 수 있는 식품의 비제한적인 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 등을 들 수 있다.The kind of the food is not particularly limited, and includes food in a conventional sense. Non-limiting examples of the food to which the substance can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice creams, , A drink, an alcoholic beverage, and a vitamin complex.
본 발명의 상기 건강 기능 식품 조성물이 음료 조성물인 경우, 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 천연 탄수화물의 비제한적인 예로 포도당, 과당과 같은 모노사카라이드; 말토스, 수크로오스와 같은 디사카라이드; 덱스트린, 사이클로덱스트린과 같은 천연 감미제; 사카린, 아스파르탐과 같은 합성 감미제 등을 들 수 있다. 상기 첨가되는 추가 성분의 비율은 당업자의 선택에 의해 적절하게 결정될 수 있다.When the health functional food composition of the present invention is a beverage composition, various flavoring agents, natural carbohydrates, and the like may be contained as additional components such as ordinary beverages. Non-limiting examples of such natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweetening agents such as dextrin, cyclodextrin; Synthetic sweetening agents such as saccharin and aspartame, and the like. The proportion of the additional component added may be appropriately determined by a person skilled in the art.
상기 외에 본 발명의 건강 기능 식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강 기능 식품 조성물은 천연 과일 주스, 과일 음료 또는 야채 음료 등의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 사용되거나 2 이상을 조합하여 사용할 수 있다. 이러한 첨가물의 비율 또한 당업자에 의해 적절히 선택될 수 있다.In addition to the above, the health functional food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloid thickening agents, pH adjusting agents, stabilizers, , Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food composition of the present invention may contain pulp for the production of natural fruit juice, fruit drink or vegetable drink. These components may be used independently or in combination of two or more. The ratios of these additives can also be suitably selected by those skilled in the art.
본 발명에서 사용되는 용어, "개선"은 치료되는 상태와 관련된 파라미터, 예를 들면 증상의 정도를 적어도 감소시키는 모든 행위를 의미한다.
As used herein, the term "improvement" means any action that at least reduces the degree of symptom associated with the condition being treated.
본 발명의 상륙 추출물 또는 이의 분획물을 함유하는 조성물은 갑상선 질환을 효과적으로 예방, 개선 또는 치료하는 효과를 나타낸다.The composition containing the landing extract or the fraction thereof of the present invention has an effect of effectively preventing, ameliorating or treating thyroid disease.
구체적으로, 본 발명의 상기 상륙 추출물 또는 이의 분획물을 함유하는 조성물은 갑상선 호르몬, 특히 티록신의 분비를 조절하는 효능이 있어, 갑상선 호르몬 조절 이상 증상을 나타내는 갑상선 질환에 특히 유용하며, 독성이 없고 천연물로부터 유래된 물질이어서 인체에 적용하여도 부작용없이 활용할 수 있는 이점이 있다.
In particular, the composition containing the above-mentioned landing extract or its fractions of the present invention has an effect of regulating the secretion of thyroid hormone, especially thyroxine, and is particularly useful for thyroid disease which exhibits abnormal thyroid hormone control. It is advantageous to apply it to human body without any side effect.
도 1은 TSH 결핍 조건에서, 상륙 추출물의 세포 증식에 대한 효과를 확인한 것으로, 도 1a는 FRTL-5 세포에 17β-estradiol (E2) (10 nM)를 처리하지 않거나 단독 처리하거나, 또는 상륙의 물 추출물, 상륙의 에탄올 추출물 10 μg/mL 또는 100 μg/mL 과 함께 처리하여 24시간 동안 5H 배지에서 배양한 후, WST를 처리하여 490nm에서 흡광도를 측정한 결과이다. 도 1b는 FRTL-5 세포에 17β-estradiol (E2) (10 nM)를 처리하지 않거나 단독 처리하거나, 또는 상륙의 물 추출물, 상륙의 에탄올 추출물 100 μg/mL과 함께 처리하여 24시간 동안 5H 배지에서 배양한 후 세포의 형태를 100배 배율의 현미경에서 관찰한 도이다. FIG. 1 shows the effect of the landing extract on cell proliferation in the presence of TSH deficiency. FIG. 1a shows the effect of treatment with 17β-estradiol (E2) (10 nM) in FRTL- Extracts, and 10 μg / mL or 100 μg / mL of the ethanol extracts from the landings, cultured in 5H medium for 24 hours, and then treated with WST to measure the absorbance at 490 nm. Figure 1B shows that FRTL-5 cells are treated with or without alone 17p-estradiol (E2) (10 nM) or treated with landing water extract, landing ethanol extract at 100 μg / ml for 24 hours in 5H medium After culturing, the morphology of the cells was observed under a microscope at a magnification of 100 times.
이하, 실시예를 통하여 본 발명을 보다 상세히 설명한다. 다만, 이들 실시예는 본 발명을 예시적으로 설명하기 위한 것에 불과하므로 본 발명의 범위가 이들 실시예에 의해 한정되는 것으로 해석되어서는 아니된다.
Hereinafter, the present invention will be described in more detail by way of examples. It should be understood, however, that these examples are for illustrative purposes only and are not to be construed as limiting the scope of the invention.
제조예Manufacturing example 1: 상륙 추출물의 제조 1: Manufacture of Landing Extracts
(1) 상륙의 물 추출물(1) Water extract of landing
시중의 상륙을 입수하여, 상륙 100 g 당 1L의 물을 가하여 환류 추출기를 이용해 100℃에서 2시간 동안 환류 추출하는 과정을 2회 반복하였다. 그 다음, 환류 추출한 결과물을 여과지로 여과한 후 회전 진공 농축기를 이용하여 농축하였다. 농축된 샘플을 물에 녹인 후 -70℃에서 10시간 동안 보관한 다음 동결 건조기를 이용하여 -40℃에서 동결 건조하여 분말로 제조하었다.
A landing on the market was obtained, and 1 L of water was added per 100 g of landing, followed by reflux extraction for 2 hours at 100 ° C using a reflux extractor. This procedure was repeated twice. Then, the resultant obtained by refluxing was filtered through a filter paper, and then concentrated using a rotary vacuum concentrator. The concentrated sample was dissolved in water, stored at -70 ° C for 10 hours, and lyophilized at -40 ° C using a freeze dryer to prepare a powder.
(2) 상륙의 에탄올 추출물(2) Landing ethanol extract
상륙 100 g 당 1L의 70% 에탄올을 가하고, 2시간 동안 소니케이션(Sonication) 처리를 한 후, 70시간 동안 침출 추출을 수행한 다음, 여과지로 여과한 후 회전 진공 농축기를 이용하여 농축하였다. 농축된 샘플을 물에 녹인 후 -70℃에서 10시간 동안 보관한 다음 동결 건조기를 이용하여 -40℃에서 동결 건조하여 분말로 제조하었다.
1 L of 70% ethanol was added per 100 g of landing, sonicated for 2 hours, leached out for 70 hours, filtered through filter paper and concentrated using rotary vacuum concentrator. The concentrated sample was dissolved in water, stored at -70 ° C for 10 hours, and lyophilized at -40 ° C using a freeze dryer to prepare a powder.
실시예Example 1: 상륙 추출물의 효능을 확인하기 위한 갑상선 세포주 배양 1: Cultivation of thyroid cell line to confirm efficacy of landing extract
랫트의 갑상선 세포주인 FRTL-5(Rat thyroid follicular cell)을 ATCC사(미국)에서 분양받아 사용하였다. FRTL-5 갑상선 세포주를, 0.5%의 신생 우 혈청(newborn calf serum) 및 1%의 페니실린-스트렙토마이신을 포함하고, 6가지 호르몬 혼합물(10 mU/ml의 TSH, 0.01 mg/ml의 인슐린, 10 nM의 하이드로코르티솔(hydrocortisone), 0.005 mg/ml의 트랜스페린(transferrin), 10 ng/ml의 소마토스탄틴(somatostatin), 10 ng/ml의 글리실-L-히스티딜-L-리신 아세테이트)이 포함된 Ham's F12 배지(이하, 6H 배지)에서, 온도 37℃, CO2 5%인 조건에서 배양기에서 배양하였다. 배양액은 23일마다 교환하였고, 세포는 57일마다 계대 배양하였다.
The rat thyroid cell line FRTL-5 (Rat thyroid follicular cell) was purchased from ATCC (USA). The FRTL-5 thyroid cell line was cultured in RPMI-16 medium containing 0.5% neonatal calf serum and 1% penicillin-streptomycin and containing 6 hormone mixtures (10 mU / ml TSH, 0.01 mg / ml insulin, 10 ml of hydrocortisone, 0.005 mg / ml transferrin, 10 ng / ml somatostatin, 10 ng / ml glycyl-L-histidyl-L-lysine acetate) Were cultured in a Ham's F12 medium (hereinafter referred to as 6H medium) at 37 DEG C and 5% CO 2 in an incubator. The culture medium was changed every 23 days, and the cells were subcultured every 57 days.
실시예Example 2: 2: TSHTSH 과잉 조건에서, 상륙 추출물 처리에 따른 티록신 분비 감소 확인 Confirmed reduction of thyroxine secretion by treatment with landing extract in excess condition
상기 제조예 1에서 제조한 상륙의 물 추출물 및 에탄올 추출물 각각을 1 mg/ml로 6H 배지에 녹인 후, 0.2 μm의 멤브레인 필터로 여과한 다음, -20℃에서 보관하였다.Each of the water extract and ethanol extract of the landing prepared in Preparation Example 1 was dissolved in 6H medium at 1 mg / ml, filtered through a 0.2 μm membrane filter, and stored at -20 ° C.
상기 실시예 1에서 배양한 FRTL-5 세포를 96 웰-조직 배양 플래이트에 웰당 1.5 ×104 세포가 되도록 분주하여 24시간 동안 배양한 후, 이를 6H 배지에 옮기고 상기에서 처리한 상륙의 물 추출물 또는 에탄올 추출물을 각각 10 μg/ml가 되도록 처리하고, 10분 간 37℃에서 배양하였다. 10분 후 1 nM의 NaI를 배지에 첨가하여 3시간 더 배양하고, 세포 배양액의 상청액을 원심 분리기로 1000 rpm으로 5분 동안 원심 분리하여 상층액을 모아 Total thyroxine(Total T4) ELISA kit(Alpha Diagnostic Intl. Inc, USA)의 각 웰에 25 μl의 standards, control 그리고 상기 상층액 시료를 넣고, 100 μl의 enzyme conjugate를 더 넣어 살짝 흔들어준 후 1시간 동안 실온에서 배양하였다.The FRTL-5 cells cultured in Example 1 were divided into a 96-well tissue culture plate at a density of 1.5 x 10 4 cells per well, cultured for 24 hours, transferred to a 6H medium, Ethanol extracts were each treated at 10 μg / ml and cultured at 37 ° C for 10 minutes. After 10 minutes, 1 nM of NaI was added to the culture medium, followed by further incubation for 3 hours. The supernatant of the cell culture was centrifuged at 1000 rpm for 5 minutes using a centrifuge to collect the supernatant. Total thyroxine (Total T4) ELISA kit 25 μl of standards, control, and supernatant samples were added to each well of a 100 μl enzyme conjugate and incubated at room temperature for 1 hour.
그 다음, 상층액을 모두 버리고, 300 μl의 증류수로 웰을 5회 반복하여 세척하였다. 그 다음, 200 μl의 HRP-substrate mix(soln. A 와 B)를 넣고 다시 실온에서 10분간 배양한 후, 50 μl의 stop solution을 넣고 450 nm에서 ELISA reader로 흡광도를 측정하였다.The supernatant was then discarded and the wells were washed five times with 300 μl of distilled water. Subsequently, 200 μl of HRP-substrate mix (soln. A and B) was added and incubated at room temperature for 10 minutes. Then, 50 μl of stop solution was added and absorbance was measured with an ELISA reader at 450 nm.
하기 표 1은 상기 실험 결과를 나타낸 것이다. 하기 표1에서 티록신의 양은 무처리 대조군에서의 티록신 양 대비 백분율로 계산한 수치이다.Table 1 below shows the results of the experiment. In Table 1 below, the amount of thyroxine is calculated as a percentage of the amount of thyroxine in the untreated control group.
*P＜0.05
* P < 0.05
상기 실험 결과, TSH이 존재하는 조건인 6H 배지에서는, 상륙 추출물을 처리한 경우 무처리 대조군에 비해 티록신 양이 감소된 것으로 확인되었다.As a result of the above experiment, it was confirmed that the amount of thyroxine was reduced in the 6H medium in the presence of TSH compared to the untreated control group when the landing extract was treated.
구체적으로, 6H 배지는 TSH가 함유되어 있어 티록신의 분비가 촉진되는 조건에 해당하는데, 상륙의 물 추출물 또는 에탄올 추출물 모두에서 티록신의 분비가 유의성있게 감소한 것으로 확인되었으며, 70% 에탄올 추출에서 티록신 감소 활성이 더욱 강하게 나타났다.Specifically, the 6H medium contained TSH, which corresponds to the condition that thyroxine secretion is promoted. It was confirmed that the secretion of thyroxine was significantly decreased in both the water extract and the ethanol extract of the landing. In 70% ethanol extraction, .
즉, 갑상선 호르몬은 뇌하수체 전엽에서 TSH가 분비되어 피드백 작용에 의해서 조절되는데, 티록신 분비가 과잉되면 TSH의 분비가 억제되어 갑상선 호르몬 분비가 억제된다. 상륙 추출물이 6H 배지의 조건에서 티록신을 증가시키지 않고 감소시킨 것은 상륙 추출물이 갑상선 호르몬의 피드백을 잘 수용하고 있다는 것을 뒷받침하는 것으로, 본 발명의 상륙 추출물이 TSH와 유사한 작용을 하여 갑상선 호르몬의 분비 조절에 효과적으로 사용될 수 있음을 보여준다.
That is, thyroid hormone is secreted from the anterior pituitary gland by TSH secretion, which is regulated by feedback. When thyroxine secretion is excessive, TSH secretion is suppressed and thyroid hormone secretion is suppressed. The reduction of the landing extract without increasing the thyroxine under the condition of 6H medium supports the fact that the landing extract has good feedback of the thyroid hormone. The landing extract of the present invention has a similar action to TSH and regulates the secretion of thyroid hormone As shown in FIG.
실시예Example 3: 3: TSHTSH 결핍 조건에서, 상륙 추출물 처리에 따른 티록신 분비 증가 확인 Confirmation of increase of thyroxine secretion by treatment of landing extract in deficient condition
상기 실시예 2에 있어서, 하기의 사항을 제외하고는 상기 실시예 2와 동일한 방법으로 실험을 수행하였다.In Example 2, the experiment was carried out in the same manner as in Example 2 except for the following.
상기 제조예 1에서 제조한 상륙의 물 추출물 및 에탄올 추출물 각각을 1 mg/ml로 5H 배지(6H 배지 중 TSH만이 불포함된 배지)에 녹인 후, 0.2 μm의 멤브레인 필터로 여과한 다음, -20℃에서 보관하였다.Each of the water extract and ethanol extract of the landing prepared in Preparation Example 1 was dissolved in 5H medium (medium containing no TSH in 6H medium) at a concentration of 1 mg / ml, filtered through a 0.2 μm membrane filter, Lt; / RTI >
상기 실시예 1에서 배양한 FRTL-5 세포를 96 웰-조직 배양 플래이트에 웰당 1.5 ×104 세포가 되도록 분주하여 24시간 동안 배양한 후, 이를 5H 배지에 옮기고 상기에서 처리한 상륙의 물 추출물 또는 에탄올 추출물을 각각 10 μg/ml가 되도록 처리하였다.The FRTL-5 cells cultured in Example 1 were added to a 96-well tissue culture plate at a rate of 1.5 × 10 4 cells per well. After the cells were cultured for 24 hours, they were transferred to 5H medium, Ethanol extracts were each treated at 10 μg / ml.
하기 표 2은 상기 실험 결과를 나타낸 것이다. 하기 표2에서 티록신의 양은 무처리 대조군에서의 티록신 양 대비 백분율로 계산한 수치이다.Table 2 shows the results of the experiment. In Table 2 below, the amount of thyroxine was calculated as a percentage of the amount of thyroxine in the untreated control group.
**P＜0.01
** P < 0.01
상기 실험 결과, TSH이 불포함된 조건인 5H 배지에서는 6H 배지에서의 결과와 정반대로, 상륙 추출물을 처리한 경우 무처리 대조군에 비해 티록신 양이 증가된 것으로 확인되었다.As a result of the above experiment, it was confirmed that the amount of thyroxine was increased in the 5H medium, which is a condition without TSH, when the landing extract was treated, as opposed to the result in the 6H medium.
구체적으로 상륙의 물 추출물 및 에탄올 추출물은 TSH가 함유되어 있지 않은 5H 배지에서 무처리 대조군에 비해 티록신의 분비를 유의성있게 증가시키는 것으로 확인되었으며, 특히 에탄올 추출물의 경우 티록신의 분비 촉진 활성이 더욱 우수하였다. 상기 결과는 갑상선 호르몬이 부족한 상태에서 본 발명의 상륙 추출물이 TSH와 유사한 작용하여 갑상선 호르몬의 분비를 조절한다는 것을 뒷받침한다.
In particular, the water extracts and ethanol extracts from the landing were found to significantly increase thyroxine secretion in the TSH-free 5H medium compared to the untreated control. In particular, the ethanol extracts were superior to thyroxine in promoting secretion of thyroxine . These results support that the landing extract of the present invention acts like TSH to regulate the secretion of thyroid hormone in the state of lack of thyroid hormone.
실시예Example 4: 4: TSHTSH 결핍 조건에서, 상륙 추출물의 세포 증식 효과 확인 Identification of the cell proliferation effect of the landing extract in the deficient condition
TSH가 포함되지 않은 배지(5H 배지)에서 상륙 추출물이 FRTL-5 세포의 증식을 유도하는지를 확인하기 위하여, WST (4-[3-(4-iodophenyl)-2-(4-nitrophenyl)-2H-5-tetrazolio]-1,3-benzene disulfonate) assay (Daeil Lab Service Co., Ltd)를 사용하였다. (4-iodophenyl) -2- (4-nitrophenyl) -2H-pyrazol-3-yl) -propionamide 5-tetrazolio] -1,3-benzene disulfonate assay (Daeil Lab Service Co., Ltd.) was used.
먼저 FRTL-5 세포를 96-웰 플레이트에 1.5×104 세포/ml씩 분주하여 6H 배지에서 24시간 동안 배양하였다. 이후, 17β-estradiol (E2) (10 nM)를 처리하지 않거나 단독 처리하거나, 또는 상륙의 물 추출물, 상륙의 에탄올 추출물 10 μg/mL 또는 100 μg/mL 과 함께 처리하여 24시간 동안 5H 배지에서 배양하였다. 이 후, WST를 2시간 동안 더 처리하고, 490nm에서 흡광도를 측정하였다. 그 결과, 세포에 상륙의 물 추출물, 상륙의 에탄올 추출물 10 μg/mL 또는 100 μg/mL를 24시간 동안 처리 후에, 상대적인 세포의 수는 농도 의존적으로 증가하였다(도 1a). 또한, E2 (10 nM)과 상륙의 물 추출물, 상륙의 에탄올 추출물을 함께 처리한 경우, 상륙 추출물에 의한 세포 증식이 더욱 증가하였다. FRTL-5 cells were first plated in 96-well plates at 1.5 x 104 cells / ml and cultured in 6H medium for 24 hours. Thereafter, 17β-estradiol (E2) (10 nM) was treated untreated or alone, or treated with landing water extract, landing ethanol extract at 10 μg / mL or 100 μg / mL and cultured in 5H medium for 24 hours Respectively. Thereafter, the WST was further treated for 2 hours and absorbance was measured at 490 nm. As a result, the relative number of cells was increased in a dose-dependent manner (Fig. 1A) after 24 h of treatment with water extract of landing water, ethanol extract of landing 10 μg / mL or 100 μg / mL of cells for 24 hours. In addition, when E2 (10 nM) and water extract of landing and ethanol extract of landing were treated together, the cell proliferation by the landing extract was further increased.
또한, TSH가 포함되지 않은 배지(5H 배지)에서 상륙 추출물이 세포 증식에 미치는 영향을 확인하기 위하여, 세포 형태를 관찰하였다. 이를 위해, FRTL-5 세포는 96-웰 플레이트에 1.5×104 세포/ml씩 분주하여 6H 배지에서 24시간 동안 배양하였다. 이후, 17β-estradiol (E2) (10 nM)를 처리하지 않거나 단독 처리하거나, 또는 상륙의 물 추출물, 상륙의 에탄올 추출물 100 μg/mL과 함께 처리하여 24시간 동안 5H 배지에서 배양한 후 세포의 형태를 100배 배율의 현미경에서 관찰하였다. 그 결과, 무처리 대조군과 비교하여, 상륙의 물 추출물, 상륙의 에탄올 추출물 100 μg/mL을 처리한 세포에서 세포의 수와 부피가 증가하였다(도 1b). 또한, E2 (10 nM)과 상륙 추출물을 함께 처리한 경우, 세포 수와 부피가 더욱 증가하였다. 상기 결과를 통해 상륙 추출물이 5H 배지에서 FRTL-5 세포의 증식을 유도함을 확인하였으며, 이러한 효과는 estradiol과 시너지 효과를 가짐을 확인하였다.
The cell morphology was also examined in order to examine the effect of the landing extract on cell proliferation in the medium containing no TSH (5H medium). For this purpose, FRTL-5 cells were cultured in 6H medium for 24 hours at a density of 1.5 × 10 4 cells / ml in a 96-well plate. Thereafter, 17β-estradiol (E2) (10 nM) was treated with or without alone, or with a water extract of landing, 100 μg / mL of ethanol extract from landing, cultured in 5H medium for 24 hours, Were observed under a microscope with a magnification of 100 times. As a result, the number and volume of cells increased in cells treated with 100 μg / mL of water extract and landing ethanol extract, compared with the untreated control group (FIG. 1B). In addition, when E2 (10 nM) and Landing extract were treated together, cell number and volume increased. From the above results, it was confirmed that the landing extract induces the proliferation of FRTL-5 cells in the 5H medium, and this effect has synergistic effect with estradiol.
실시예Example 5: 상륙 추출물의 세포 독성 확인 5: Cytotoxicity of Landing Extracts
상륙 추출물의 세포 독성 여부를 확인하기 위하여 다음과 같은 실험을 수행하였다.The following experiment was carried out to confirm cytotoxicity of the landing extract.
24-웰 플레이트에 FRTL-5 세포를 1.5×104 세포/ml씩 분주하여 24시간 동안 배양한 후, 10 g/ml 내지 100 g/ml의 농도로 희석시킨 상기 제조예 1에서 제조한 상륙 추출물이 첨가된 새 배지로 교체하여 다시 24시간 동안 배양하였다. 그 다음, 상기 배지에 MTT(3-(4,5-디메틸티아졸-2-일)-2,5-디페닐테트라졸륨 브로마이드) 5 mg/ml를 첨가한 후 37℃, 5% CO2 배양기에서 2시간 동안 배양한 다음, 형성된 포마잔(fomazan)을 DMSO로 녹이고, 96 웰 플레이트로 옮긴 후 570 nm에서 ELISA reader로 흡광도를 측정하였다.FRTL-5 cells were seeded at a concentration of 1.5 x 10 4 cells / ml in a 24-well plate, cultured for 24 hours, and then treated with the Landing extract prepared in Preparation Example 1, which was diluted to a concentration of 10 g / ml to 100 g / ml And the cells were cultured again for 24 hours. Subsequently, 5 mg / ml of MTT (3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) was added to the medium and incubated at 37 ° C in a 5% CO 2 incubator For 2 hours, the formed fomazan was dissolved in DMSO, transferred to a 96-well plate, and absorbance was measured with an ELISA reader at 570 nm.
하기 표 3은 상기 실험 결과를 나타낸 것이다.Table 3 below shows the results of the experiment.
상기 실험 결과, 상륙의 물 추출물 및 에탄올 추출물을 처리했을 때, 세포 생존율이 50%가 되는 농도와 갑상선 호르몬 조절 활성을 나타내는 농도를 비교하면, 갑상선 호르몬 조절 활성을 나타내는 농도에서는 세포 독성은 나타내지 않는 것으로 확인되었다.
As a result of the above experiment, when the water extract and the ethanol extract of the landing were treated, the cytotoxicity was not shown at the concentration showing the thyroid hormone controlling activity when the cell survival rate was 50% and the thyroid hormone controlling activity was compared .
실시예Example 6: 상륙 추출물의 급성 독성 확인- 6: Identification of acute toxicity of landing extract - inin vivovivo
상륙 추출물의 급성 독성 여부를 확인하기 위하여 다음과 같은 실험을 수행하였다.The following experiment was carried out to confirm the acute toxicity of the landing extract.
실험 동물로 6주령의 특정 병원체 부재(specific pathogen-free, SPF) SD계 랫트를 사용하여 급성 독성 실험을 수행하였다. 각군당 2 마리씩의 랫트에 제조예 1에서 제조한 상륙 추출물을 각각 0.5% 메틸셀룰로즈 용액에 현탁하여 10g/kg/㎖의 용량으로 1회 단회 경구투여 하였다. 시험 물질 투여 후 랫트의 폐사 여부, 임상 증상, 체중 변화를 관찰하고 혈액학적 검사 및 혈액 생화학적 검사를 실시하였으며, 부검하여 육안으로 복강 장기 및 흉강 장기의 이상 여부를 관찰하였다.
Acute toxicity tests were carried out using specific pathogen-free (SPF) SD rats at 6 weeks of age as experimental animals. Each of the two rats in each group was suspended in 0.5% methylcellulose solution, and each of them was orally administered once at a dose of 10 g / kg / ml. After the administration of the test substance, death, clinical symptoms and weight change of the rats were observed, and hematological test and blood biochemical test were performed, and autopsy was performed to observe whether or not the abdominal organs and thoracic organs were abnormal.
상기 실험 결과, 상륙의 물 추출물 또는 에탄올 추출물을 투여한 모든 랫트에서 특기할 만한 임상 증상이나 폐사된 동물은 없었으며, 체중 변화, 혈액 검사, 혈액 생화학 검사, 부검 소견 등에서도 독성 변화는 관찰되지 않았다.As a result of the above experiment, no rat clinical symptoms or dead animals were observed in all of the rats administered with the water extract of the landing or the ethanol extract, and no toxic change was observed in weight change, blood test, blood biochemical test, .
구체적으로, 본 발명의 상륙 추출물은 모두 랫트에서 10g/kg 까지도 독성 변화를 나타내지 않았으며, 경구 투여 최소 치사량(LD50)은 추출물 10g/kg 이상으로 판단되었다. 랫트의 복강에 상륙의 70% 에탄올 추출물(에탄올은 증발시켜 없앰)을 주사한 경우의 LD50은 18g/kg이었다.Specifically, the landing extract of the present invention did not show any toxic change up to 10 g / kg in rats, and the oral dose of LD 50 was determined to be 10 g / kg or more of the extract. LD 50 was 18 g / kg when 70% ethanol extract of landing in abdominal cavity of rats was injected (ethanol was not evaporated).
따라서, 본 발명의 상륙 추출물은 1회에 지나치게 과량으로 투여하지 않는 한, 생체 내에서 독성을 일으키지 않는 것으로 확인되었다
Therefore, it was confirmed that the landing extract of the present invention does not cause toxicity in vivo unless it is administered in excess amount once at a time
본 명세서는 본 발명의 기술 분야에서 통상의 지식을 가진 자이면 충분히 인식하고 유추할 수 있는 내용은 그 상세한 기재를 생략하였으며, 본 명세서에 기재된 구체적인 예시들 이외에 본 발명의 기술적 사상이나 필수적 구성을 변경하지 않는 범위내에서 보다 다양한 변형이 가능하다. 따라서 본 발명은 본 명세서에서 구체적으로 설명하고 예시한 것과 다른 방식으로 실시될 수 있으며, 이는 본 발명의 기술 분야에 통상의 지식을 가진 자이면 이해할 수 있는 사항이다.It is to be understood that both the foregoing general description and the following detailed description of the present invention are exemplary and explanatory and are intended to provide further explanation of the invention as claimed. More variations are possible within a range that does not. Accordingly, the present invention may be embodied in other ways than those specifically described and illustrated herein, and it may be understood by those skilled in the art.
Claims (12)
Landing ( Phytolacca esculenta Houttuyn) extract or fractions thereof as an active ingredient.
The pharmaceutical composition according to claim 1, wherein the landing extract is obtained by extracting landing with water, a C 1 to C 4 alcohol, or a mixed solvent thereof.
3. The pharmaceutical composition according to claim 2, wherein the landing extract is obtained by extracting landing with water or ethanol.
The pharmaceutical composition according to claim 1, wherein the landing extract is obtained by extracting roots, stems or leaves of landing.
The pharmaceutical composition according to claim 1, wherein the landing extract or the fraction thereof is to prevent or treat thyroid disease by controlling secretion of thyroxine.
The pharmaceutical composition according to claim 1, wherein the thyroid disease is selected from the group consisting of hyperthyroidism, hypothyroidism, thyroiditis, thyroid nodule, and thyroid cancer.
A method for preventing or treating a thyroid disease, comprising administering the pharmaceutical composition of any one of claims 1 to 6 to a subject other than a human.
Landing ( Phytolacca esculenta Houttuyn) extract or fractions thereof as an active ingredient for preventing or ameliorating a thyroid disease.
The food composition according to claim 8, wherein the landing extract is obtained by extracting landing with water, a C 1 to C 4 alcohol, or a mixed solvent thereof.
The food composition according to claim 9, wherein the landing extract is obtained by extracting landing with water or ethanol.
9. The food composition according to claim 8, wherein the landing extract is obtained by extracting roots, stems or leaves of landing.
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| KR20190099958A (en) * | 2018-02-20 | 2019-08-28 | 한국식품연구원 | Composition for Preventing, Improving or Treating of muscular disease containing Phytolacca esculenta extract |
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| KR20120131956A (en) | 2011-05-27 | 2012-12-05 | 연세대학교 산학협력단 | A Composition for Treating or Preventing Thyroid Disorders |
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| KR20120131956A (en) | 2011-05-27 | 2012-12-05 | 연세대학교 산학협력단 | A Composition for Treating or Preventing Thyroid Disorders |
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| KR20190099958A (en) * | 2018-02-20 | 2019-08-28 | 한국식품연구원 | Composition for Preventing, Improving or Treating of muscular disease containing Phytolacca esculenta extract |
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