KR101814132B1 - Composition for Treatment of Lung Cancer and Functional Food Comprising Extract of Patriniae Radix - Google Patents
Composition for Treatment of Lung Cancer and Functional Food Comprising Extract of Patriniae Radix Download PDFInfo
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- KR101814132B1 KR101814132B1 KR1020110027371A KR20110027371A KR101814132B1 KR 101814132 B1 KR101814132 B1 KR 101814132B1 KR 1020110027371 A KR1020110027371 A KR 1020110027371A KR 20110027371 A KR20110027371 A KR 20110027371A KR 101814132 B1 KR101814132 B1 KR 101814132B1
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- lung cancer
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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Abstract
본 발명은 패장근 추출물의 신규한 용도에 관한 것에 관한 것으로서, 보다 상세하게는 패장근 에탄올 추출물을 유효성분으로 함유하는 폐암 예방 및 치료용 조성물 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 패장근 에탄올 추출물을 유효성분으로 함유하는 폐암 예방용 기능성 식품에 관한 것이다. 본 발명에 따른 폐암 치료용 조성물 및 기능성 식품은 폐암 세포의 성장을 억제하고 세포사멸을 유도하는 효과가 있어 폐암 치료 및 예방에 효과적으로 사용할 수 있다.
More particularly, the present invention relates to a composition for the prevention and treatment of lung cancer, which contains an extract of L. longissimus as an active ingredient, Ethanol extract as an active ingredient. The composition for treating lung cancer and the functional food according to the present invention have an effect of inhibiting the growth of lung cancer cells and inducing apoptosis, and thus can be effectively used for treatment and prevention of lung cancer.
Description
본 발명은 패장근 추출물의 신규한 용도에 관한 것이다. 구체적으로, 본 발명은 폐암에 대해 탁월한 예방 또는 치료 효능을 나타내는 패장근 추출물을 유효성분으로 함유하는 치료용 조성물 및 기능성 식품에 관한 것이다.
The present invention relates to a novel use of L. root extract. Specifically, the present invention relates to a therapeutic composition and a functional food containing an extract of Lycopersicon esculentum (Lycopersicon esculentum) showing an excellent prophylactic or therapeutic efficacy against lung cancer as an active ingredient.
현대인의 주요 질환 중에서, 암의 치료방법과 진단방법에 관한 연구는 발병빈도가 높은 폐암, 간암, 위암 등을 중심으로 진행되고 있다. 폐암이란 폐에 생긴 악성 종양을 말하며, 크게 암세포가 기관지, 세기관지, 폐포 등 폐를 구성하는 조직에서 처음 발생한 원발성 폐암과 암세포가 다른 기관에서 생겨나 혈관이나 림프관을 타고 폐로 이동해 증식하는 전이성 폐암으로 나눌 수 있다.Among the major diseases of modern people, studies on the methods of treatment and diagnosis of cancer are proceeding mainly on lung cancer, liver cancer and stomach cancer which have a high incidence. Lung cancer is a malignant tumor that occurs in the lungs and can be divided into metastatic lung cancer in which primary cancerous lung cancer and cancer cells first appear in tissues constituting the lungs such as bronchi, bronchioli, alveoli and the like, have.
2009년에 발표된 한국중앙암등록본부 자료에 의하면 2007년에 우리나라에서는 연 평균 161,920건의 암이 발생되었는데, 그 중 폐암은 남녀를 합쳐서 연 평균 17,846건으로 전체 암 발생의 11.0%로 4위를 차지하였다. 인구 10만명당 조발생률은 36.3건이다. 남녀의 성비는 3.51:1로 남자에게서 더 많이 발생하였다. 발생건수는 남자가 연 평균 12,841건으로 남성의 암 중에서 2위를 차지하였고, 여자는 연 평균 5,005건으로 여성의 암 중에서 5위를 차지하였다. 남녀를 합쳐서 본 연령대별로는 70대가 34.1%로 가장 많고, 60대가 31.3%, 50대가 14.8%의 순이다(보건복지가족부 중앙암등록본부 2009년 12월 21일 발표 자료).According to the data released by the Korea Central Cancer Registry in 2009, 161,920 cases of cancer were found annually in Korea, among which 17,846 cases of lung cancer were combined with men and women, accounting for 11.0% . The incidence rate per population of 100,000 is 36.3. The sex ratio of male and female was 3.51: 1 and more occurred in male. The average incidence of males was 12,841 males and females ranked second in males and fifth females with 5,005 a year. According to this age group, the age group showed the highest rate of 34.1% in the 70s, followed by 31.3% in the 60s and 14.8% in the 50s (published by Dec. 21, 2009, Ministry of Health, Welfare and Family Affairs Central Cancer Registry).
폐암은 현미경적으로 암세포의 크기와 형태에 따라 비소세포 폐암(non-small cell lung cancer: NSCLC)과 소세포 폐암(small cell lung cancer: SCLC)으로 구분된다. 이렇게 비소세포 폐암과 소세포 폐암을 구분하는 것은 임상적 경과와 치료가 다르기 때문인데, 전체 폐암의 80~85%를 차지하는 비소세포 폐암은 조기에 진단할 경우 수술적 요법으로 치료할 수 있으나, 조기 발견이 어려운 폐암의 특성상 늦게 발견되어 치료가 어려운 경우가 많다. 비소세포 폐암은 3종류의 서브 타입으로 구성된다: 40% 선암(adenocarcinoma), 40% 편평상피세포암(squamous cell carcinoma) 및 20% 대세포암(large cell carcinoma). TMN 병기 구분법 (staging system)이 폐암의 관리에 널리 받아들여지고 있다. 일차 종양은 종양 크기, 부위 및 국부적 병발(local involvement)에 따라 4개의 T 카테고리 (T1-T4)로 구분된다. 림프 절 확산(spread)은 폐 내의 기관지/폐(bronchio/pulmonary) 내로 전달(N1), 상기 일차 종양과 같은 측면 상의 종격동 확산(medistinal spread)(N2) 및 상기 일차 폐 종양의 맞은 편으로 종격동 확산 또는 상부클라비움 병발(supraclavilcular involvement)(N3)로 구분된다. 원격 또는 전이 확산(metastatic spread)은 없거나 있다(M0 또는 M1). 일반적으로 전이가 이루어지지 않은 폐암은 외과적 수술을 통하여 제거하는 방법으로 치료를 한다. 그러나, 폐암 제거 수술 후의 재발율은 20 내지 50%로 높다(Cancer: Principles & Practice of Oncology, 56th. ed. In: Devita DV, Hellman S, Rosenberg SA, eds. Philadelphia, PA: Lippincott Williams & Wilkins, 2001).Lung cancer is divided microscopically into non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) according to the size and shape of cancer cells. This is because the differentiation between non-small cell lung cancer and small cell lung cancer is due to the different clinical course and treatment. Non-small cell lung cancer, which accounts for 80% to 85% of all lung cancers, The nature of difficult lung cancer is often found late and difficult to treat. Non-small cell lung cancer is composed of three subtypes: 40% adenocarcinoma, 40% squamous cell carcinoma, and 20% large cell carcinoma. The TMN staging system is widely accepted in the management of lung cancer. Primary tumors are divided into four T categories (T1-T4) according to tumor size, site and local involvement. The lymphocyte spread is transmitted through the bronchio / pulmonary in the lung (N1), mediastinal spread (N2) on the same side as the primary tumor and mediastinal diffusion Or supraclavilcular involvement (N3). There is no remote or metastatic spread (M0 or M1). Generally, non-metastatic lung cancer is treated by surgical removal. However, the recurrence rate after lung cancer removal surgery is as high as 20 to 50% (Cancer: Principles & Practice of Oncology, 56th ed., Devita DV, Hellman S, Rosenberg SA, eds. Philadelphia, PA: Lippincott Williams & ).
특히 비소세포성 폐암은 치료 후 재발률이 높고, 높은 발병률에 비하여 기존의 항암제를 이용한 화학적 치료에 대한 내성이 쉽게 발생하여 효과적인 치료가 어려운 것으로 알려져 있다. 상기 비소세포성 폐암의 경우, 최근의 암 치료법의 발달에도 불구하고 10년 생존률이 10% 이하로 매우 낮다. 따라서 이러한 비소세포성 폐암에 대한 치료제 개발이 매우 중요하다.
In particular, non-small cell lung cancer has a high recurrence rate after treatment and is resistant to chemotherapy using conventional anticancer agents as compared with a high incidence, and it is known that effective treatment is difficult. In the case of non-small cell lung cancer, the 10-year survival rate is as low as 10% or less despite the recent development of cancer therapy. Therefore, development of therapeutic agents for such non-small cell lung cancer is very important.
패장근(Patriniae Radix)은 마타리라고도 하며, 쌍떡잎식물 합판화군 꼭두서니목 마타리과(Valerianaceae) 마타리속(Patrinia Juss.)의 여러해살이 풀로, 일본 열도의 북쪽부터 남으로 타이완, 중국 및 시베리아 동부에 분포하며, 높이 60~150 ㎝ 내외이고 뿌리줄기는 굵으며 옆으로 뻗는다. 마타리속은 전세계에 약 15종이 있으며 우리나라에는 약 4종인 마타리, 돌마타리, 금마타리, 뚝깔이 있다. 곧게 자라는 줄기에 마주나는 잎은 깃꼴겹잎으로 깊게 갈라지며 가장자리에 톱니가 있다. 8~10월에 줄기와 가지 끝의 산방꽃차례에 자잘한 노란색 꽃이 촘촘히 모여 핀다. 꽃은 여름부터 가을에 걸쳐서 피고 노란색이다. 연한 순을 나물로 이용하고 전초를 소염, 어혈 또는 고름 빼는 약으로 사용한다. Patriniae Radix , also known as matari , is a perennial herbaceous plant belonging to the family Madrinidae ( Valrinanaceae ) of the dicotyledonous division of the dicotyledonous plant. It is distributed in the Taiwan, China and eastern Siberia from north to south of the Japanese archipelago, It is 60 ~ 150 ㎝ in height and rootstock is thick and extends to the side. There are about 15 species of martari in the world, and there are four kinds of martari, stone martari, gold matsuri, Leaves that face on straight growing stem are deeply split into double leaf, with sawtooth on edge. In August ~ October, small yellow flowers bloom densely in the flower buds of the stem and end of branches. Flowers bloom from summer to autumn and are yellow. It uses soft seeds as herbs and uses the outpost as an anti-inflammatory, eosinophilic or pus-subtracting medicine.
성분은 뿌리에는 사포닌이 있다. 뿌리의 용혈지수는 1:500, 사포닌의 용혈지수는 1:50,000이다. 지금까지 4개의 사포닌, 즉 파트리니오시드 A, B, C, D가 분리되었는데, 주성분을 이루는 것은 파트리니오시드 D이다. 이 사포닌올레오놀산 C30H48O3을 아글루콘으로 하고 포도당, 아라비노오스, 크실로오스로 이루어졌다. 이밖에 정유 약 8%, 휘발성산 약 1.5%, 흔적의 알칼로이드가 있다. 전초에서는 사포닌(용혈지수 1:1,600~2,860)과 흔적의 알칼로이드가 확인되었다. 뚝갈뿌리에는 모노페르펜 배당체인 로가닌, 빌로시드, 모르로니시드가 있다.The ingredient has saponin in the root. The root hemolysis index is 1: 500 and the saponin hemolysis index is 1: 50,000. To date, four saponins have been isolated, namely, the parturinosides A, B, C and D, the main component being partirionosid D. This saponin oleanolic acid C 30 H 48 O 3 was composed of agglutinin, glucose, arabinose and xylose. In addition, there are about 8% of essential oil, about 1.5% of volatile acid, and trace alkaloids. Saponin (hemolysis index 1: 1,600 ~ 2,860) and traces of alkaloid were detected in the outpost. There are monoglyceride, bilocide, and moronicide monopelpene glycosides in the root.
작용은 뿌리줄기와 뿌리는 동물 실험과 임상에서 바구니나물뿌리와 비슷한 작용이 있으며 치료 효과는 그보다 더 좋은 것으로 밝혀졌다. 파트리니오시드는 다른 사포닌과 마찬가지로 용혈작용, 국소자극작용이 있다. 뚝갈전초 추출물은 간세포의 재생을 촉진하고 간세포변성을 막는다. 응용은 바구니나물과 같다. 동의치료에서 염증약, 배농약, 정혈약으로 종창과 부기, 산후 배아픔 등 부인과에서 주로 쓴다.The effects of rootstocks and roots are similar to those of baskets in animal experiments and clinics, and the therapeutic effect was found to be even better. Like other saponins, it has hemolytic and topical stimulants. Chrysanthemum cepa extract promotes hepatocyte regeneration and prevents hepatocyte degeneration. The application is like a basket of herbs. It is mainly used in gynecological medicine such as inflammation medicine, pesticide, hypertension medicine swelling and swelling and postpartum abdominal pain.
그러나, 아직까지 어떠한 문헌에서도 패장근의 폐암 억제 효과에 대해서는 알려진 바가 없다.
However, the efficacy of L-carnitine to inhibit lung cancer is unknown in any literature.
이에, 본 발명자들은 패장근에 대한 생약 연구를 하던 중, 패장근 추출물이 폐암 세포를 효과적으로 사멸시킬 수 있음을 확인함으로써 본 발명을 완성하였다.
Accordingly, the inventors of the present invention completed the present invention by confirming that the extract of L. japonica can effectively kill lung cancer cells while conducting herbal medicine for L. japonica.
본 발명의 목적은 패장근 추출물을 유효성분으로 함유하는 폐암 치료용 조성물 및 기능성 식품을 제공하는 것이다.
It is an object of the present invention to provide a composition for treating lung cancer and a functional food containing an extract of Lycopersicon esculentum as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 패장근(Patriniae Radix) 에탄올 추출물을 유효성분으로 함유하는 폐암 예방 및 치료용 조성물을 제공한다.In order to achieve the above object, the present invention provides a composition for preventing and treating lung cancer, which comprises an extract of Patriniae Radix ethanol as an active ingredient.
또한, 본 발명은 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 패장근(Patriniae Radix) 에탄올 추출물을 유효성분으로 함유하는 폐암 예방용 기능성 식품을 제공한다.
In addition, the present invention provides a functional food for lung cancer prevention containing an extract of Patriniae radix ethanol which comprises a food-acceptable food-aid additive as an active ingredient.
이하, 본 발명을 상세히 설명한다. Hereinafter, the present invention will be described in detail.
본 발명은 패장근(Patriniae Radix) 에탄올 추출물을 유효성분으로 함유하는 폐암 예방 및 치료용 조성물을 제공한다. 본 발명의 조성물은 활성성분으로서 패장근 추출물을 포함하며, 추가적으로 약제학적으로 허용가능한 담체 또는 희석제를 포함할 수 있다.The present invention provides a composition for preventing and treating lung cancer, which comprises an extract of Patriniae radix ethanol as an active ingredient. The composition of the present invention comprises Lycopersicon esculentum extract as an active ingredient and may further comprise a pharmaceutically acceptable carrier or diluent.
본 발명의 폐암 예방 및 치료용 조성물에 있어서, 상기 에탄올 추출물은 50℃에서 24시간 동안 추출되는 것이 바람직하고, 이때 상기 에탄올 추출물은 45℃ 감압 조건에서 건조 및 농축되는 것이 보다 바람직하며, 상기 에탄올은 95%인 것이 가장 바람직하다.In the composition for preventing and treating lung cancer according to the present invention, it is preferable that the ethanol extract is extracted at 50 ° C. for 24 hours, and the ethanol extract is more preferably dried and concentrated under reduced pressure at 45 ° C., Most preferably 95%.
또한, 본 발명의 폐암 예방 및 치료용 조성물에 있어서, 상기 폐암은 비소세포 폐암인 것이 바람직하고, 이때 상기 비소세포 폐암은 선암(adenocarcinoma), 편평상피세포암(squamous cell carcinoma) 및 대세포암(large cell carcinoma)으로 이루어진 군중에서 선택된 암종에 의하여 유발되는 것이 보다 바람직하다.
In the composition for preventing and treating lung cancer according to the present invention, the lung cancer is preferably non-small cell lung cancer, wherein the non-small cell lung cancer is adenocarcinoma, squamous cell carcinoma and large cell carcinoma and large cell carcinoma).
본 발명에서, 사용된 용어 "폐암(lung cancer)"은 폐에 발생하는 악성 종양을 의미하며, 소세포 폐암과 선암, 편평상피세포암, 대세포 폐암 등의 비소세포 폐암을 모두 포함하는 의미이다.In the present invention, the term "lung cancer " as used herein means a malignant tumor arising in the lung, including all small cell lung cancer, non-small cell lung cancer such as adenocarcinoma, squamous cell cancer, and large cell lung cancer.
상기 "약제학적으로 허용가능한 담체"는 신체의 한 기관 또는 부분으로부터 신체의 다른 기관 또는 부분으로 활성 성분을 수송하는 역할을 하는 액체 또는 고체 충진제, 희석제, 부형제 또는 용매와 같은 약제학적으로 허용되는 물질, 조성물 또는 운반체(vehicle)를 의미한다.The term "pharmaceutically acceptable carrier" is intended to encompass pharmaceutically acceptable substances, such as liquid or solid fillers, diluents, excipients or solvents, which serve to transport the active ingredient from one or more parts of the body to other organs or parts of the body , Composition or vehicle.
본 발명의 폐암 치료용 조성물은 유효성분과 함께 추가로 약제학적으로 허용되는 1종 이상의 담체를 첨가하여 약제로 제조할 수 있다. 상기 담체로는 식염수, 완충 식염수, 물, 글리세롤 및 에탄올 등이 있으나 이에 한정되지 않으며, 당해 기술 분야에 알려진 적합한 제제(Remingtons's Pharmaceutical Science(최근판), Mack Publishing Company, Easton PA)를 모두 사용 가능하다.The composition for treating lung cancer of the present invention may be prepared by adding one or more pharmaceutically acceptable carriers together with an active ingredient. Such carriers include, but are not limited to, saline, buffered saline, water, glycerol, and ethanol, and any suitable formulation known in the art (Remington's Pharmaceutical Science (recent edition), Mack Publishing Company, Easton PA) .
본 발명의 패장근 추출물을 약제화하기 위한 제제는 임상 투여시에 경구로 투여가 가능하며 일반적인 의약품 제제의 형태로 사용될 수 있으며, 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 경구를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드, 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제 감미제, 방향제, 보존제 등이 포함될 수 있다.The pharmaceutical composition of the present invention can be administered orally in clinical administration and can be used in the form of a general pharmaceutical preparation. In the case of formulation, the filler, extender, binder, wetting agent, , A surfactant, and the like. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like. Examples of liquid formulations for oral use include suspensions, solutions, emulsions and syrups. Common diluents such as water, In addition to liquids and paraffins, various excipients such as wetting agent sweetening agents, perfumes, preservatives and the like may be included.
또한, 본 발명의 조성물에 추가될 수 있는 생약재는 약학적으로 허용되는 임의의 생약재일 수 있으며, 예를 들면, 고본(Angelicae tenuissimae Radix), 천마(Gastrodiae Rhizoma), 시호(Bapleuri Radix), 당귀(Angelicae gigantis Radix), 도인(Persicae Semen), 계지(Cinnamomi Ramulus), 대황(Rhei Rhizoma), 감초(Glycyrrhizae Radix), 천궁(Cnidii Rhizoma), 진피(Aurantii nobilis Pericarpium), 택사(Alismatis Rhizoma), 황련(Coptidis Rhizoma), 황금(Scutellariae Radix), 복령(Hoelen), 작약(Paeoniae Radix), 백출(Atractylodis Rhizoma alba), 황백(Phellodendri Cortex), 치자(Gardeniae Fructus), 반하(Pinelliae Tuber), 조구등(Uncaria Ramuluset Uncus), 지실(Ponciri Fructus), 인삼(Gingseng), 맥문동(Liriopis Tuber), 원지(Polygalae Radix), 석창포(Acori graminei Rhizoma), 창출(Atractylodis Rhizoma alba), 감국(Chrysanthemi Flos), 방풍(Ledebouriellae Radix), 생강(Zingiberis Rhizoma crudus), 망초(Natrii sulfas), 대조(Zizyphi Fructus), 단삼(Salviae Radix), 목단피(Mautan Radicis Cortex), 지황(Rehmanniae Radix), 박하(Menthae Herba), 산약(Dioscoreae Rhizoma), 저령(Polyporus), 하수오(Polygonimultiflori Radix), 구자(Allii tuberosi Semen), 결명자(Cassiae Semen), 구기자(Lycii Fructus), 독활(Araliae cordatae Radix), 두충(Eucommiae Cortex), 백화사설초(Hedyotis Herba), 삼백초(Saururus Herba), 인진(Artemisiaecapillaris Herba), 지모(Anemarrhenae Rhizoma), 홍화(Carthami Flos), 황기(Astragali Radix), 석송자(Lycopodium), 은행잎(Ginkgonis Folium), 황정(Polygonati Rhizoma), 연자육(Nelumbinis Semen), 용골(Fossilia ossis Mastodi), 지골피(Lycii radicis Cortex), 우슬(Achyranthis Radix), 숙지황(Rehmanniae Radix preparata), 흑임자(Perillae Semen), 백자인(Thujae Semen), 맥아(Hordei Fructus germinatus), 토사자(Cuscutae Semen), 파극천(Morindae Radix), 해송(Pini koraiensis Radix) 등을 단독으로 또는 배합하여 사용할 수 있다.
In addition, the herbal medicine which may be added to the composition of the present invention may be any pharmaceutically acceptable herbal medicine, for example, Angelicae tenuissimae Radix, Gastrodiae Rhizoma, Bapleuri Radix, (Rhizoma), Angelicae gigantis Radix, Persicae Semen, Cinnamomi Ramulus, Rhei Rhizoma, Glycyrrhizae Radix, Cnidii Rhizoma, Aurantii nobilis Pericarpium, Alismatis Rhizoma, Coptidis Rhizoma, Scutellariae Radix, Hoelen, Paeoniae Radix, Atractylodis Rhizoma alba, Phellodendri Cortex, Gardeniae Fructus, Pinelliae Tuber, Uncaria Ramuluset, Uncus, Ponciri Fructus, Ginseng, Liriopis Tuber, Polygalae Radix, Acori graminei Rhizoma, Atractylodis Rhizoma alba, Chrysanthemi Flos, Ledebouriellae Radix, ), Ginger (Zingiberis Rhizoma crudus), ganoderma (Natrii sulfas), control yphi Fructus, Salviae Radix, Mautan Radicis Cortex, Rehmanniae Radix, Menthae Herba, Dioscoreae Rhizoma, Polyporus, Polygonimultiflori Radix, Allii tuberosi Semen, Cassiae Semen, Lycii Fructus, Araliae cordatae Radix, Eucommiae Cortex, Hedyotis Herba, Saururus Herba, Artemisiaecapillaris Herba, Anemarrhenae, Rhizoma, Rhizoma, Carthami Flos, Astragali Radix, Lycopodium, Ginkgonis Folium, Polygonati Rhizoma, Nelumbinis Semen, Fossilia ossis Mastodi, Lycii radicis Cortex ), Achyranthis Radix, Rehmanniae Radix preparata, Perillae Semen, Thujae Semen, Hordei Fructus germinatus, Cuscutae Semen, Morindae Radix, Pini koraiensis Radix) may be used alone or in combination.
본 발명의 조성물은 실제 임상 투여시에 비경구의 여러 가지 제형으로 투여될 수 있는데, 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드, 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제 감미제, 방향제, 보존제 등이 포함될 수 있다. 구체적으로, 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성용제, 현탁용제로는 프로필렌글리콜(Propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. 또한 치료제의 효능 증진을 위해 칼슘이나 비타민 D3를 첨가할 수 있다. 이러한 조성물은 단위-용량(1회분) 또는 다중-용량(수 회분) 용기, 예를 들면, 밀봉된 앰풀 및 바이알에 제시될 수 있고, 사용 직전에 멸균성 액상 담체, 예를 들면, 주사용 수의 부가 만을 요구하는 동결-건조 조건 하에 저장할 수 있다. 즉석의 주사 용제 및 현탁제는 멸균성 산제, 과립제 및 정제로부터 제조할 수 있다.The composition of the present invention can be administered in various forms of parenteral administration at the time of actual clinical administration. Solid formulations include tablets, pills, powders, granules, capsules and the like. Examples of the liquid formulations include suspensions, , Syrups, and the like. Various excipients such as wetting agent sweeteners, fragrances, preservatives, etc. may be included in addition to simple diluents commonly used in water, liquids and paraffins. Specifically, formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like can be used as the non-aqueous solvent and suspension agent. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like. Calcium or vitamin D 3 may also be added to enhance the efficacy of the therapeutic agent. Such compositions may be presented in unit-dose (one-time) or multi-dose (several-dose) containers, such as sealed ampoules and vials, and may be presented in a sterile liquid carrier, Lt; RTI ID = 0.0 > freeze-drying < / RTI > Immediate injectable solutions and suspensions may be prepared from sterile powders, granules and tablets.
본 발명의 제제는 대상의 연령, 성별, 상태, 체내에서 활성 성분의 흡수도, 불활성율 및 배설속도, 병용되는 약물에 따라 달리 적용될 수 있다. 본 발명은 또한 투약 단위의 제형들을 포함한다. 제형은 개별 투약 형태, 예를 들면 정제, 피복 정제, 캡슐제, 환제, 좌약 및 앰플제로 존재하고, 약제 중 유효 화합물의 함량은 개별 투약량의 분율 또는 배수에 해당한다. 투약 단위는, 예를 들면 개별 투여량의 1, 2, 3 또는 4배로, 또는 1/2, 1/3 또는 1/4배를 함유할 수 있다. 개별 투여량은 바람직하기로는 유효 화합물이 1회에 투여되는 양을 함유하며, 이는 통상 1일 투여량의 전부, 1/2, 1/3 또는 1/4배에 해당한다.
The formulations of the present invention may be applied differently depending on the age, sex, condition of the subject, the degree of absorption of the active ingredient in the body, the rate of inactivation and the rate of excretion, and the drugs used in combination. The present invention also includes formulations of dosage units. The formulations are presented in separate dosage forms, such as tablets, coated tablets, capsules, pills, suppositories, and ampoules, and the content of active compound in the drug is a fraction or multiple of the individual dosage. Dosage units may contain, for example, 1, 2, 3 or 4 times, or 1/2, 1/3 or 1/4 times the individual doses. The individual doses preferably contain amounts in which the active compound is administered in a single dose, which usually corresponds to the full, half, one-third or one-fourth of the daily dose.
본 발명에서 용어, "추출물(extract)"은 천연물로부터 분리된 활성성분을 의미한다. 추출물은 물, 유기용매, 또는 이의 혼합용매를 이용하는 추출과정으로 획득할 수 있으며, 추출액, 이의 건조 분말 또는 이를 이용하여 제형화된 모든 형태를 포함한다.
As used herein, the term "extract " means an active ingredient isolated from a natural product. The extract can be obtained by an extraction process using water, an organic solvent, or a mixed solvent thereof, and includes an extract, a dry powder thereof, or all the forms formulated with it.
본 발명의 구체적인 실시에서, 패장근의 에탄올 추출물은 A549 비소세포성 폐암 세포를 100 ㎍/㎖에서 98.1% 사멸시키고, EC50(half maximal effective concentration)은 63.7 ㎍/㎖이었다. 상기 결과는 본 발명의 패장근 추출물이 우수한 A549 비소세포성 폐암 세포의 사멸 활성을 가지며, 나아가 폐암 치료 및 예방 활성을 가진다는 것을 입증하는 것이다.
In a specific embodiment of the present invention, the ethanol extract of L. japonica killed 98.1% of A549 non-small cell lung cancer cells at 100 μg / ml and the EC 50 (half maximal effective concentration) was 63.7 μg / ml. The above results demonstrate that the extracts of the present invention have good killing activity against A549 non-small cell lung cancer cells and further have a therapeutic and preventive activity against lung cancer.
본 발명에서 용어, "예방"이란 조성물의 투여로 폐암 발병을 억제 또는 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" means any act that inhibits or delays the onset of lung cancer by administration of the composition.
본 발명에서 용어, "치료"란 조성물의 투여로 폐암의 증세가 호전되거나 이롭게 변경하는 모든 행위를 의미한다.The term "treatment" in the present invention means all the actions of improving or alleviating symptoms of lung cancer by administration of the composition.
본 발명에서 패장근 추출물은 물, 유기 용매, 또는 이의 혼합 용매를 사용하여 추출하여 사용할 수 있다. 바람직하게는 유기 용매, 특히 에탄올을 사용하여 추출한다. 추출한 액은 바로 사용하거나 또는 농축 및/또는 건조하여 사용할 수 있다. 유기용매를 사용하여 추출하는 경우, 메탄올, 에탄올, 이소프로판올, 부탄올, 에틸렌, 아세톤, 헥산, 에테르, 클로로포름, 에틸아세테이트, 부틸아세테이트, 디클로로메탄, N, N-디메틸포름아미드(DMF), 디메틸설폭사이드(DMSO), 1,3-부틸렌글리콜, 프로필렌글리콜 또는 이들의 혼합용매인 유기용매를 사용하며 생약의 유효 성분이 파괴되지 않거나 최소화된 조건에서 실온 또는 가온하여 추출할 수 있다. 추출하는 유기용매에 따라 약제의 유효성분의 추출정도와 손실정도가 차이가 날 수 있으므로, 알맞은 유기용매를 선택하여 사용하도록 한다. 추출 방법은 특별히 제한되지 않고, 예를 들어 냉침 추출, 초음파 추출, 환류 냉각 추출 등이 있다. 여과는 추출액으로부터 부유하는 고체 입자를 제거하는 과정으로, 면, 나일론 등을 이용하여 입자를 걸러내거나 한외여과, 냉동여과법, 원심분리법 등을 사용할 수 있으나 이에 제한되지 않는다.In the present invention, the extract may be extracted using water, an organic solvent, or a mixed solvent thereof. Preferably an organic solvent, especially ethanol. The extracted liquid can be used directly or by concentrating and / or drying. In the case of extraction using an organic solvent, an organic solvent such as methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF) (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof. The active ingredient of the herbal medicine may be extracted at room temperature or warmed under the condition that the active ingredient is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and the degree of loss of the active ingredient of the medicament may differ. Therefore, an appropriate organic solvent should be selected and used. The extraction method is not particularly limited, and examples thereof include cold extraction, ultrasonic extraction, and reflux cooling extraction. Filtration is a process of removing suspended solid particles from an extract, and may be performed by filtering particles using a cotton, nylon, or the like, or by ultrafiltration, freezing filtration, centrifugation, and the like.
추출액의 농축에는 감압농축, 역삼투압 농축 등의 방법이 사용될 수 있다. 농축 후 건조 단계는 동결건조, 진공건조, 열풍건조, 분무건조, 감압건조, 포말건조, 고주파건조, 적외선건조 등을 포함하나 이에 제한되지 않는다. 경우에 따라, 최종 건조된 추출물을 분쇄하는 공정을 추가할 수 있다.Concentration of the extract can be carried out by reduced-pressure concentration, reverse osmosis concentration, or the like. The post-concentration drying step includes, but is not limited to, freeze drying, vacuum drying, hot air drying, spray drying, vacuum drying, foam drying, high frequency drying, infrared drying and the like. In some cases, a step of pulverizing the final dried extract may be added.
또한, 상기 추출물은 추가의 분획 공정을 수행할 수 있다. 바람직하게는 상기 추출물을 증류수에 현탁시켜 비극성 유기 용매, 예를 들어, 헥산, 에테로, 디클로로메탄, 클로로포름, 에틸아세테이드 또는 이들의 혼합 용매로 비극성용매 가용층을 추출, 분리하여 수득하도록 하고, 이를 농축 및/또는 건조하여 사용할 수 있다.In addition, the extract may be subjected to an additional fractionation process. Preferably, the extract is suspended in distilled water, and the non-polar solvent soluble layer is extracted and separated by using a nonpolar organic solvent such as hexane, ether, dichloromethane, chloroform, ethyl acetate, or a mixed solvent thereof , Which can be used by concentration and / or drying.
본 발명에서, 용어 "약제학적으로 허용가능한 염"이란 약리학적 또는 생리학적으로 허용되는 무기산, 유기산 및 염기로부터 유도된 염을 의미한다. 적합산 산의 예로는 염산, 브롬산, 황산, 질산, 과염소산, 푸마르산, 말레산, 인산, 글리콜산, 락트산, 살리실산, 숙신산, 톨루엔-p-설폰산, 타르타르산, 아세트산, 시트르산, 메탄설폰산, 포름산, 벤조산, 말론산, 나프탈렌-2-설폰산, 벤젠설폰산 등을 포함할 수 있다. 적합한 염기로부터 유도된 염은 알칼리 금속, 예들 들어, 나트륨, 알칼리토금속, 예들 들어, 마그네슘, 암모늄 등을 포함할 수 있다.
In the present invention, the term "pharmaceutically acceptable salt" means salts derived from pharmacologically or physiologically acceptable inorganic acids, organic acids and bases. Examples of suitable acids include hydrochloric, hydrobromic, sulfuric, nitric, perchloric, fumaric, maleic, phosphoric, glycolic, lactic, salicylic, succinic, toluene- Formic acid, benzoic acid, malonic acid, naphthalene-2-sulfonic acid, benzenesulfonic acid, and the like. Salts derived from suitable bases may include alkali metals, such as sodium, alkaline earth metals, such as magnesium, ammonium, and the like.
본 발명의 폐암 질환 예방 및 치료용 약학조성물은 조성물 총 중량에 대하여 상기 추출물 또는 화합물을 0.1 내지 50 중량%로 포함한다. 또한, 상기 조성물은 약효를 증가시키지는 않으나 약재 조성물에 통상 사용되어 냄새, 맛, 시각 등을 향상시킬 수 있는 추가성분을 포함할 수 있다. 또한, 상기 조성물은 비타민 B1, B2, B6, C, E, 니아신, 카르니친, 베타인, 엽산 판토텐산, 비오틴, 아연, 철, 칼슘, 크롬, 마그네슘, 이들의 혼합물 등의 무기, 유기 첨가물들을 추가로 포함할 수 있다. 또한, 상기 조성물은 단독 사용하거나 기존 사용되어진 폐암에 대한 치료 활성을 가지는 물질을 포함할 수 있다.
The pharmaceutical composition for prevention and treatment of lung cancer diseases according to the present invention comprises 0.1 to 50% by weight of the extract or the compound, based on the total weight of the composition. In addition, the composition does not increase the efficacy, but may contain additional ingredients that are commonly used in pharmaceutical compositions to improve odor, taste, visual appearance, and the like. In addition, the composition may further comprise inorganic and organic additives such as vitamins B1, B2, B6, C, E, niacin, carnitine, betaine, folic acid pantothenic acid, biotin, zinc, iron, calcium, chromium, magnesium, As shown in FIG. In addition, the composition may contain a substance which has therapeutic activity on lung cancer which has been used alone or has been used.
본 발명에서 용어, "환자"는 폐암 및 이의 직, 간접적 원인에 의해 유발된 질환을 가지고, 본 발명의 조성물을 투여에 의하여 증상이 호전될 수 있는 인간과 말, 양, 돼지, 염소, 낙타, 영양, 개 등의 동물을 의미한다. 본 발명의 패장근 추출물을 포함하는 조성물을 환자에게 투여함으로써, 상기에서 언급한 폐암을 효과적으로 예방 및 치료할 수 있다. 본 발명의 조성물을 기존의 폐암 치료제와 병행하여 투여할 수 있다.The term "patient" in the present invention refers to a disease caused by lung cancer and its direct or indirect cause, and can be administered to humans and horses, sheep, pigs, goats, camels, Nutrition, and dogs. The above-mentioned lung cancer can be effectively prevented and treated by administering to a patient a composition comprising the L. elm extract of the present invention. The composition of the present invention may be administered in combination with a conventional lung cancer therapeutic agent.
본 발명에서 용어, "투여"는 어떠한 적절한 방법으로 환자에게 소정의 물질을 도입하는 것을 의미하며, 본 발명의 조성물의 투여 경로는 목적 조직에 도달할 수 있는 한 어떠한 일반적인 경로를 통하여 경구 또는 비경구 투여될 수 있다. 또한, 조성물은 활성 물질이 표적 세포로 이동할 수 있는 임의의 장치에 의해 투여될 수 있다.The term "administering" in the present invention means introducing a predetermined substance into a patient in any appropriate manner, and the administration route of the composition of the present invention may be oral or parenteral ≪ / RTI > The composition may also be administered by any device capable of transferring the active agent to the target cell.
본 발명의 조성물은 약제학적으로 유효한 양으로 투여한다. 본 발명에서 용어, "약제학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효 용량 수준은 환자의 성병, 연령, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료 기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 알려진 요소에 따라 결정될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래의 치료제와는 순차적 또는 동시에 투여될 수 있다. 단일 또는 다중 투여될 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. 본 발명의 제조 방법에 따라 제조된 추출물 또는 화합물을 포함하는 조성물의 투여방법은 경구투여 또는 정맥투여가 바람직하고, 일반적으로 그 유효 용량은 경구투여인 경우에는 보통 성인을 기준으로 1회에 1 내지 500 ㎎/㎏이 바람직하며, 정맥투여인 경우에는 1 내지 100 ㎎/㎏이 바람직하며, 하루 2-3 회 투여될 수 있다. 특정 환자에 대한 투여용량 수준은 성별, 연령, 건강상태, 식이, 투여시간, 투여방법, 약제혼합, 환자의 상태 및 신경 질환의 발병 정도에 따라 변화될 수 있다.
The composition of the present invention is administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the sex of the patient, age, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. May be administered singly or multiply. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without adverse effect, and can be easily determined by those skilled in the art. The method for administration of the composition comprising the extract or the compound prepared according to the preparation method of the present invention is preferably oral or intravenous. Generally, the effective dose is usually 1 to 10 times, 500 mg / kg is preferable, and in case of intravenous administration, 1 to 100 mg / kg is preferable, and it can be administered 2-3 times a day. The dosage level for a particular patient can be varied depending on the sex, age, health condition, diet, time of administration, method of administration, drug mix, patient condition, and severity of neurological disease.
또한, 본 발명은 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 패장근(Patriniae Radix) 에탄올 추출물을 유효성분으로 함유하는 폐암 예방용 기능성 식품을 제공한다.In addition, the present invention provides a functional food for lung cancer prevention containing an extract of Patriniae radix ethanol which comprises a food-acceptable food-aid additive as an active ingredient.
본 발명의 기능성 식품은 에탄올 추출물을 총중량에 대해 0.1 내지 5중량% 포함하는 것이 바람직하며, 1중량%로 포함하는 것이 더욱 바람직하다. 본 발명의 기능성 식품은 그 제형에 있어서 특별히 한정되는 바는 없으며, 예를 들어 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.The functional food of the present invention preferably contains 0.1 to 5% by weight, more preferably 1% by weight, of the ethanol extract in the total weight. The functional food of the present invention is not particularly limited in its formulation and can be used in various food products such as dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramie noodle, Various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and includes all the health foods in the ordinary sense.
본 발명에 있어서 "기능성 식품"은 건강기능식품에 관한 법률 제6727호에 따른 인체에 유용한 기능성을 가진 원료나 성분을 사용하여 제조 및 가공한 식품을 의미하며, "기능성"이라 함은 인체의 구조 및 기능에 대하여 영양소를 조절하거나 생리학적 작용 등과 같은 보건 용도에 유용한 효과를 얻을 목적으로 섭취하는 것을 의미한다.
In the present invention, the term "functional food" means a food prepared and processed by using a raw material or ingredient having useful functionality in the human body according to Law No. 6727 on health functional foods, and " And function of the nutrient for the purpose of obtaining a beneficial effect in health use such as controlling the nutrient or physiological action.
본 발명에서는 A549 비소세포성 폐암 세포에 본 발명의 패장근 추출물을 투입한 결과, A549 비소세포성 폐암 세포가 사멸하는 것을 확인하였다(도 1 참조). 따라서, 패장근 추출물이 폐암 치료 효과가 있다는 것을 새롭게 알 수 있다. 이에, 본 발명에서는 패장근 추출물을 유효성분으로 함유하는 폐암 예방용 기능성 식품을 제조함으로써 본 발명을 완성하였다(제조예 2 참조)
In the present invention, A549 non-small cell lung cancer cells were killed by A549 non-small cell lung cancer cells (see FIG. 1 ). Therefore, it can be newly found that L. japonica extract is effective in treating lung cancer. Thus, the present invention has completed the present invention by preparing a functional food for lung cancer prevention containing an extract of Lycopersicon esculentum as an active ingredient (see Production Example 2 )
상기에서 살펴본 바와 같이, 본 발명의 패장근 추출물은 폐암 세포의 성장을 억제하고 세포사멸을 유도한다. 따라서 본 발명에 따른 폐암 치료용 조성물은 폐암 환자의 치료에 매우 효과적일 것이다.
As described above, the L. elm extract of the present invention inhibits the growth of lung cancer cells and induces apoptosis. Therefore, the composition for treating lung cancer according to the present invention will be very effective for the treatment of lung cancer patients.
도 1은 인간 폐암 세포인 A549 비소세포성 폐암 세포에서 패장근의 도입이 폐암 세포의 성장에 미치는 영향을 알아보기 위한 Alamar Blue 분석 결과이고, 이때 X축은 패장근 추출물의 농도이고, Y축은 생존한 인간 폐암 세포의 생존율을 나타낸다. FIG. 1 shows the results of Alamar Blue analysis to examine the effect of the introduction of L-carnitine on the growth of lung cancer cells in human lung cancer cell A549 non-small cell lung cancer cells, wherein the X-axis is the concentration of the L. root extract and the Y- Indicating the survival rate of human lung cancer cells.
이하, 본 발명을 하기 실시예에 의거하여 보다 상세하게 설명하고자 한다. 단, 하기 실시예는 본 발명을 예시하기 위한 것일 뿐, 본 발명은 하기 실시예에 의해 한정되는 것이 아니고, 본 발명의 기술적 사상을 벗어나지 않는 범위 내에서 치환 및 균등한 타 실시예로 변경할 수 있음은 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자에게 있어서 명백할 것이다.
Hereinafter, the present invention will be described in more detail based on the following examples. It should be noted, however, that the following examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The present invention is not limited to the following examples. Will be apparent to those skilled in the art to which the present invention pertains.
<실시예 1> 패장근 추출물의 제조<Example 1> Preparation of extracts of L. japonica
서울 약재상에서 구입한 패장근(중국산) 3 ㎏을 음지 및 실온에서 5일간 건조하고 분쇄하였다. 상기 분쇄된 패장근을 95% 에탄올(ethanol) 30 ℓ에 침지시키고 50℃에서 24시간 동안 추출하였다. 이것을 여과지를 통하여 여과한 후 45℃ 감압 조건에서 건조 및 농축하여 총 추출물 411 g을 수득하고, -20℃에서 보관하였다.
3 kg of L. jangun (Chinese origin) purchased from Seoul Medicinal Plant was dried and pulverized for 5 days at the shade and room temperature. The ground pulverulent muscle was immersed in 30 L of 95% ethanol and extracted at 50 DEG C for 24 hours. This was filtered through a filter paper, dried and concentrated under reduced pressure at 45 ° C to obtain 411 g of a total extract, which was stored at -20 ° C.
<실시예 2> 패장근 추출물이 폐암 세포의 성장에 미치는 영향<Example 2> Effect of Lactobacillus root Extract on Growth of Lung Cancer Cells
상기 실시예 1에서 추출한 패장근 추출물이 폐암 세포의 성장에 미치는 영향을 알아보기 위하여, 인간 폐암 세포인 A549 비소세포성 폐암 세포에 패장근의 에탄올 추출물을 48시간 처리하고 Alamar Blue 분석을 시행하였다. Alamar Blue 분석은 MTT 분석의 변형된 형태인데, 특정 효소에 의해서 분해되는 화합물을 살아있는 세포에 처리한 후 화합물이 분해되면서 나오는 생성물의 형광 세기를 측정함으로써 약물을 처리한 후 살아있는 세포의 상대적인 숫자를 알아내는 실험방법이다. 하기에서 보다 상세히 설명한다.
To investigate the effect of L. elm extract on the growth of lung cancer cells, A549 non-small cell lung cancer cells were treated with Alamar Blue for 48 hours. The Alamar Blue assay is a modified form of MTT assay that measures the fluorescence intensity of a product after degradation of a compound that is degraded by a specific enzyme into a living cell to determine the relative number of living cells after treatment It is an experimental method. This will be described in more detail below.
<2-1> 인간 폐암 세포주의 준비 및 처리<2-1> Preparation and treatment of human lung cancer cell line
본 발명에 사용된 폐암 세포주인 A549 세포는 한국세포주은행(Korean Cell Line Bank, KCLB)으로부터 분양받아 실험에 이용하였다. 구체적으로, A549 폐암 세포를 10% FBS(fetal bovine serum, 소태아혈청)(Welgene)와 25 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)를 포함하는 RPMI 1640 배지(Robwell Park Memorial Institute에 의하여 연구 개발된 부유배양용 배지)에서 계대배양하였다.
A549 cells, the lung cancer cell line used in the present invention, were purchased from Korean Cell Line Bank (KCLB) and used for the experiments. Specifically, A549 lung cancer cells were cultured in RPMI 1640 medium (Robwell Park Memorial) containing 10% fetal bovine serum (Welgene) and 25 mM HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid) ≪ RTI ID = 0.0 > Institute). ≪ / RTI >
<2-2> A549 비소세포성 세포의 세포 성장 억제 측정<2-2> Measurement of cell growth inhibition of A549 non-small cell
상기 실시예 1에서 추출한 패장근 추출물이 폐암 세포인 A549 비소세포성 세포 성장 억제효과를 확인하였다. 구체적으로, 96 웰 플레이트에 각 웰 당 1.4 X 103 개의 A549 폐암 세포를 주입(seeding)하고 24시간 동안 배양한 후, DMSO(Dimethyl sulfoxide)에 녹인 상기 패장근 에탄올 추출물을 각각 0 내지 100 ㎍/㎖ 농도(구체적으로, 각각 0, 3.125, 6.25, 12.5, 25, 50 및 100 ㎍/㎖ 농도)로 48시간 동안 처리하였을 때, 세포 성장을 저해하는 정도를 확인하였다(표 1). 각 농도의 추출물을 처리한 후, 96-웰 플레이트에서 각 웰에 채워진 0.2 ㎖의 세포 배양액에 20 ㎕의 Alamar Blue 시약을 첨가한 후 플레이트를 인큐베이터에서 2시간 동안 배양하였다. 각 웰의 세포를 고르게 반응시키기 위하여 플레이트를 천천히 흔들고, 544 ㎚의 파장에서 조사광을 조사하면서 590 ㎚에서 형광의 세기를 형광광도계(Fluorescence Microplate Reader; Molecular Devices Corp.)로 측정하였고, 폐암 세포의 생존율을 도 1에 나타내었다.
The extracts of L. elicitor from Example 1 inhibited A549 non-small cell growth, a lung cancer cell. Specifically, 1.4 X 10 3 A549 lung cancer cells were seeded on a 96-well plate and cultured for 24 hours. Then, the cells were cultured in DMSO (dimethyl sulfoxide) Ethanol extract was treated for 48 hours at a concentration of 0 to 100 μg / ml (specifically, 0, 3.125, 6.25, 12.5, 25, 50 and 100 μg / ml, respectively) (Table 1). After treating each concentration of the extract, 20 μl of Alamar Blue reagent was added to 0.2 ml of the cell culture solution filled in each well in a 96-well plate, and the plate was cultured in an incubator for 2 hours. The plate was gently shaken to react cells in each well, and the intensity of fluorescence was measured at 590 nm using a fluorescence photometer (Molecular Devices Corp.) while irradiating light with a wavelength of 544 nm. Survival rate is shown in Fig.
그 결과, 표 1 및 도 1에서 나타난 바와 같이, 패장근의 처리 농도가 높을수록 폐암 세포의 성장이 감소하였으며, 이로부터 패장근이 폐암 치료 효과를 가짐을 알 수 있었다. 즉, 3.125 ㎍/㎖에서 2.7%, 6.25 ㎍/㎖에서 6.7%, 12.5 ㎍/㎖에서 5.3%, 25 ㎍/㎖에서 8.9%, 50 ㎍/㎖에서 11.1%, 100 ㎍/㎖에서 98.1%로 폐암 세포를 사멸시켰다. 아울러, EC50(half maximal effective concentration)은 63.7 ㎍/㎖로 측정되었다. 상기 표 1에서 패장근을 처리하지 않은 대조군의 폐암 세포의 생존율 수를 1을 기준으로 하여 각각의 패장근 처리 농도에 따른 48시간 후의 폐암 세포의 상대적 세포수를 기재하였다. 이와 같이, 본 발명의 패장근 추출물은 우수한 A549 비소세포성 폐암 세포 사멸 활성을 가지며, 나아가 폐암 치료 및 예방 활성을 가진다는 것을 입증한다.
As a result, as shown in Table 1 and FIG. 1, the higher the treatment concentration of the callus root, the less the growth of lung cancer cells, and it was found that the callosal root was effective in the treatment of lung cancer. In other words, it was 2.7% at 3.125 占 퐂 / ml, 6.7% at 6.25 占 퐂 / ml, 5.3% at 25 占 퐂 / ml, 8.9% at 12 占 퐂 / ml, 11.1% at 100 占 퐂 / Lung cancer cells. In addition, EC 50 (half maximal effective concentration) was measured to be 63.7 / / ml. In Table 1, the relative number of cells of lung cancer cells after 48 hours was shown based on the number of survival rate of lung cancer cells in the control group not treated with L. Thus, the L. elm extract of the present invention has excellent A549 non-small cell lung cancer cell killing activity, further demonstrating that it has lung cancer treatment and prevention activity.
<실시예 3> 패장근 추출물에 의한 급성독성 시험≪ Example 3 > Acute Toxicity Test with Extract of Lycopersicon esculentum
본 발명에 이용된 패장근은 널리 약재로 이용되고 있어서 안정성에 문제가 없을 것으로 판단하였으나, 경구 투여시 및 복강내 투여시의 독성 실험을 수행하여 이를 확인하고자 하였다.The inventors of the present invention have determined that the disintegrator used in the present invention is widely used as a medicinal agent, so that there is no problem in stability.
6주령의 특정병원부재(SPF) SD계 랫트를 사용하여 급성독성실험을 실시하였다. 군당 2 마리씩의 동물에 본 발명의 실시예 1의 패장근 추출물을 각각 0.5% 메틸셀룰로즈 용액에 현탁하여 5 g/㎏의 용량으로 단회 경구투여하였다. 시험물질 투여후 동물의 폐사여부, 임상증상, 체중변화를 관찰하고 혈액학적 검사와 혈액생화학적검사를 실시하였으며, 부검하여 육안으로 복강장기와 흉강장기의 이상여부를 관찰하였다.Acute toxicity tests were carried out using 6-week-old SPF SD rats. The extracts of Example 1 of the present invention were suspended in a 0.5% methylcellulose solution and administered to a 2-group animal at a dose of 5 g / kg in a single oral dose. After the administration of the test substance, the mortality, clinical symptoms, and weight changes of the animals were observed, and hematological tests and blood biochemical tests were carried out, and autopsy was performed to observe the abnormalities of the abdominal organs and thoracic organs.
시험결과, 시험물질을 투여한 모든 동물에서 특기할 만한 임상증상이나 폐사된 동물은 없었으며, 체중변화, 혈액검사, 혈액생화학 검사, 부검소견 등에서도 독성변화는 관찰되지 않았다. 이상의 결과 패장근 추출물은 모두 랫트에서 5 g/㎏까지 독성변화를 나타내지 않으며 경구 투여 최소치사량 (LD50)은 5 g/㎏이상인 안전한 물질로 판단되었다.
As a result of the test, there were no clinically symptomatic or dead animals in all the animals to which the test substance was administered, and no toxic change was observed in weight change, blood test, blood biochemical test, and autopsy findings. As a result, the extracts of Root Root Extract did not show any change in toxicity up to 5 g / kg in rats and it was judged that the minimum LDL (LD 50 ) was over 5 g / kg.
<제조예 1> 패장근 추출물을 유효성분으로 함유하는 폐암 치료제의 제조<Preparation Example 1> Preparation of a therapeutic agent for lung cancer containing L. japonica extract as an active ingredient
본 발명자들은 상기 실시예를 통해 패장근 추출물의 폐암 치료 효능이 뛰어남을 확인하여 패장근 추출물을 유효성분으로 함유하는 폐암 치료제를 하기와 같이 제조하였다. 또한, 하기 치료제의 제조예는 치료제 뿐만 아니라 건강식품의 제조에도 응용하여 사용될 수 있다.
The inventors of the present invention confirmed that the extract of L. japonica was excellent in the treatment of lung cancer and thus, a therapeutic agent for lung cancer containing L. japonica extract as an active ingredient was prepared as follows. In addition, the preparation examples of the following therapeutic agents can be applied not only to therapeutic agents but also to the production of health foods.
<1-1> 패장근 추출물을 함유하는 연질캅셀(soft gelatin capsules)≪ 1-1 > Soft gelatin capsules containing Lycopersicon esculentum Extract
패장근 추출물 20%Root Root Extract 20%
비타민 C 4.5%Vitamin C 4.5%
비타민 D3 0.001%Vitamin D 3 0.001%
황산망간 0.1%Manganese sulfate 0.1%
밀납 10%Wax 10%
팜유 25%Palm oil 25%
홍화씨유 30.399%
Safflower oil 30.399%
<1-2> 패장근 추출물을 함유하는 정맥주사용 제제의 제조 ≪ 1-2 > Preparation of an intravenous formulation containing L. japonica extract
패장근 추출물 0.2%Root Root Extract 0.2%
만니톨 0.3%Mannitol 0.3%
생리식염수 9.5%
Physiological saline 9.5%
<1-3> 패장근 추출물을 함유하는 정제(tablet)≪ 1-3 > Tablets containing Lycopersicon esculentum Extract
패장근 추출물 35%Lilium root extract 35%
비타민 C 10%Vitamin C 10%
비타민 D3 0.001%Vitamin D 3 0.001%
황산망간 0.1%Manganese sulfate 0.1%
결정셀룰로오즈 25.0%Crystalline cellulose 25.0%
유당 17.999%Lactose 17.999%
스테아린산마그네슘 2%
Magnesium stearate 2%
<제조예 2> 패장근 추출물을 유효성분으로 함유하는 기능성 식품의 제조Preparation Example 2 Preparation of Functional Foods Containing Lentinus edodes Extracts as Active Ingredients
본 발명자들은 상기 실시예를 통해 패장근 추출물이 폐암 치료 활성이 뛰어남을 확인하여 이를 유효성분으로 함유하는 기능성 식품을 하기와 같이 제조하였다.
The inventors of the present invention confirmed that the extracts of L. elder were excellent in lung cancer therapeutic activity through the above examples, and prepared functional foods containing the active ingredients as follows.
<2-1> 음료의 제조<2-1> Production of beverage
꿀 522 ㎎Honey 522 mg
치옥토산아미드 5 ㎎5 mg < RTI ID = 0.0 >
니코틴산아미드 10 ㎎Nicotinic acid amide 10 mg
염산리보플라빈나트륨 3 ㎎3 mg of sodium riboflavin hydrochloride
염산피리독신 2 ㎎Pyridoxine hydrochloride 2 mg
이노시톨 30 ㎎Inositol 30 mg
오르트산 50 ㎎Orthoic acid 50 mg
패장근 추출물 0.48 ~ 1.28 ㎎L. japonica extract 0.48 ~ 1.28 ㎎
물 200 ㎖200 ml of water
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 음료를 제조하였다.
A beverage was prepared using the above-mentioned composition and content by a conventional method.
<2-2> 츄잉껌의 제조<2-2> Production of chewing gum
껌베이스 20 %
설탕 76.36 ~ 76.76 %Sugar 76.36 ~ 76.76%
패장근 추출물 0.24 ~ 0.64 %L. japonica extract 0.24 ~ 0.64%
후르츠향 1 %Fruit flavor 1%
물 2 %Water 2%
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 츄잉껌을 제조하였다.
Chewing gum was prepared using the above-mentioned composition and content by a conventional method.
<2-3> 캔디의 제조<2-3> Manufacture of candy
설탕 50 ~ 60 %Sugar 50 to 60%
물엿 39.26 ~ 49.66 %Syrup 39.26 ~ 49.66%
패장근 추출물 0.24 ~ 0.64 %L. japonica extract 0.24 ~ 0.64%
오렌지향 0.1 %Orange fragrance 0.1%
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 캔디를 제조하였다.
The composition and the content of the candy were prepared using a conventional method.
<2-4> 비스켓의 제조<2-4> Production of biscuit
박력1급 88 ㎏First class 88 ㎏ power
중력1급 76.4 ㎏Gravity Class 1 76.4 kg
정백당 16.5 ㎏16.5 kg
식염 2.5 ㎏Salt 2.5 ㎏
포도당 2.7 ㎏Glucose 2.7 kg
팜쇼트닝 40.5 ㎏Palm shortening 40.5 kg
암모 5.3 ㎏Ammonium 5.3 kg
중조 0.6 ㎏0.6 kg
중아황산나트륨 0.55 ㎏Sodium bisulfite 0.55 kg
쌀가루 5.0 ㎏Rice powder 5.0 kg
비타민 B1 0.003 ㎏Vitamin B1 0.003 kg
비타민 B2 0.003 ㎏Vitamin B2 0.003 kg
밀크향 0.16 ㎏Milk flavor 0.16 kg
물 71.1 ㎏Water 71.1 kg
전지분유 4 ㎏Whole milk powder 4 ㎏
대용분유 1 ㎏1 kg of substitute milk powder
제일인산칼슘 0.1 ㎏Calcium phosphate 0.1 kg
살포염 1 ㎏Spray salt 1 kg
분무유 25 ㎏Spray oil 25 kg
패장근 추출물 0.2 ~ 0.5 ㎏Root Root Extract 0.2 ~ 0.5 ㎏
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 비스켓을 제조하였다.
The biscuits were prepared using the above-mentioned composition and content by a conventional method.
<2-5> 아이스크림의 제조<2-5> Production of ice cream
유지방 10.0 %Fat milk 10.0%
무지유고형분 10.8 %Solid oil content 10.8%
설탕 12.0 %Sugar 12.0%
물엿 3.0 %Starch syrup 3.0%
유화안정제(스팬, span) 0.5 %Emulsion stabilizer (span) 0.5%
향료(스트로베리) 0.15 %Perfume (Strawberry) 0.15%
물 63.31 ~ 62.91 %Water 63.31 ~ 62.91%
패장근 추출물 0.24 ~ 0.64 %L. japonica extract 0.24 ~ 0.64%
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 아이스크림을 제조하였다.
Ice cream was prepared using the above-mentioned composition and content by a conventional method.
<2-6> 쵸코렛의 제조<2-6> Manufacture of Chocolate
설탕 34.36 ~ 34.76 %Sugar 34.36 ~ 34.76%
코코아 버터 34 %Cocoa Butter 34%
코코아 매스 15 %Cocoa mass 15%
코코아 파우다 15 %Cocoa powder 15%
레시틴 0.5 %Lecithin 0.5%
바닐라향 0.5 %Vanilla flavor 0.5%
패장근 추출물 0.24 ~ 0.64 %L. japonica extract 0.24 ~ 0.64%
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 초코렛을 제조하였다.
The composition and the content thereof were used to prepare chocolate by a conventional method.
한편, 본 발명의 구체적 범위는 상기 기술한 실시예 보다는 특허청구범위에 의하여 한정지어지며, 특허청구 범위의 의미와 범위 및 그 등가적 개념으로 도출되는 모든 변경 및 변형된 형태를 본 발명의 범위로 포함하여 해석하여야 한다.It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory and are intended to provide further explanation of the invention, and are not intended to limit the scope of the invention. .
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