KR20120111125A - Composition for treatment of renal cell carcinoma and functional food comprising extract of cannabis semen - Google Patents
Composition for treatment of renal cell carcinoma and functional food comprising extract of cannabis semen Download PDFInfo
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- KR20120111125A KR20120111125A KR1020110029445A KR20110029445A KR20120111125A KR 20120111125 A KR20120111125 A KR 20120111125A KR 1020110029445 A KR1020110029445 A KR 1020110029445A KR 20110029445 A KR20110029445 A KR 20110029445A KR 20120111125 A KR20120111125 A KR 20120111125A
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- extract
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- kidney cancer
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- active ingredient
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Abstract
Description
본 발명은 마자인 추출물의 신규한 용도에 관한 것이다. 구체적으로, 본 발명은 신장암에 대해 탁월한 예방 또는 치료 효능을 나타내는 마자인 추출물을 유효성분으로 함유하는 치료용 조성물 및 기능성 식품에 관한 것이다.
The present invention relates to a novel use of madin extract. Specifically, the present invention relates to a therapeutic composition and functional food containing as soon as the active ingredient extract showing excellent prophylactic or therapeutic efficacy against kidney cancer.
현대인의 주요 질환 중에서, 암의 치료방법과 진단방법에 관한 연구는 발병빈도가 높은 폐암, 간암, 위암 등을 중심으로 진행되고 있다.Among the major diseases of modern people, researches on the treatment and diagnosis of cancer are being conducted mainly on lung cancer, liver cancer, and stomach cancer, which have high incidence.
신장은 혈액을 여과하여 소변을 생성함으로써 생체 내의 노폐물을 체외로 배설하는 역할을 갖는 중요한 비뇨기계 기관이다. 또한, 동시에 혈압을 조절하는 안지오텐신, 적혈구 조혈 인자인 에리스로포이에틴 등의 호르몬을 생산하는 중요한 내분비 기관이기도 하다. 신장에 발생하는 종양에는, 성인에게서 발생하는 신세포암(kindney cell cancer)과 소아에게서 발생하는 윌름스(Wilms) 종양, 및 보기 드문 종양으로서 육종(sarcoma)이 있지만, 이하에서는 가장 발생률이 높은 악성 종양인 신세포암을 신장암이라고 부른다. 신장암의 발생 빈도는 인구 10만명당 2.5명 정도이고, 남녀비는 2 내지 3:1로 남성에게 많은 경향이 있다. 비뇨기과계 악성 종양 중에서는, 전립선암, 방광암에 이어서 많은 종양이다.The kidney is an important urinary system that has the role of excreting waste products in vitro by filtering blood to produce urine. At the same time, it is also an important endocrine organ that produces hormones such as angiotensin, which regulates blood pressure, and erythropoietin, a red blood cell hematopoietic factor. Renal tumors include kidney cell cancer in adults, Wilms' tumor in children, and sarcoma as a rare tumor, but the most common malignancy below Renal cell cancer, a tumor, is called kidney cancer. The incidence of renal cancer is about 2.5 per 100,000 population, and male to female ratio is 2 to 3: 1. Among urological malignancies, there are many tumors following prostate cancer and bladder cancer.
신장암의 위험 인자로서는 유전학적 요인도 알려져 있지만, 일반적으로는 흡연, 과도한 지방 섭취 등을 들 수 있다. 또한, 장기간 투석을 받고 있는 환자에게서 상기 종양의 발생률이 높은 것으로 알려져 있다. 신장암에서는 종양의 최대 직경이 5 ㎝ 이하인 경우에는 어떠한 자각 증상이 있는 경우가 드물고, 검진시의 CT 스캔 등에 의해 발견되는 경우가 많다. 크기가 큰 종양에서는 혈뇨, 복부 종양, 동통 등이 나타날 수 있다. 또한, 전신적 증상으로서 발열, 체중 감소, 빈혈 등을 초래하는 경우가 있으며, 드물게는 내분비 인자에 의해 적혈구 증다증이나 고혈압, 고칼슘혈증 등이 발생되는 경우가 있다. 한편, 신장암의 하대정맥 내로의 진전에 의해 복부 체표의 정맥의 노장(怒張)이나 정소 정맥류가 발생하는 경우가 있다. 신장암의 약 2할은, 폐나 골 전이로부터 발견된다. 신장암에서는 정맥 중에 종양이 확산되는 경향이 강하고, 다른 장기로의 전이가 발생하기 쉽다.Genetic factors are also known as risk factors for kidney cancer, but generally include smoking and excessive fat intake. It is also known that the incidence of such tumors is high in patients undergoing long-term dialysis. In kidney cancer, when the tumor has a maximum diameter of 5 cm or less, some subjective symptoms rarely occur and are often found by CT scan or the like at the time of examination. Larger tumors may include hematuria, abdominal tumors, and pain. In addition, systemic symptoms may cause fever, weight loss, anemia and the like, and rarely, endocrine factors may cause erythrocytosis, hypertension, hypercalcemia, and the like. On the other hand, there may be cases in which vein veins of the abdominal body surface or venous varicose veins occur due to the progression of kidney cancer into the inferior vena cava. About 20% of kidney cancers are found from lung or bone metastases. In kidney cancer, tumors tend to spread in veins, and metastasis to other organs is likely to occur.
신장암의 검사법으로서는 초음파 검사, CT 검사, 혈관 조영 검사 등이 있지만, 특이적인 생화학 마커는 알려져 있지 않기 때문에 기기를 이용한 검진이 필요로 된다. 또한, 신장암은 병리학적으로 몇 개의 분류가 가능하지만, 그 중 약 90 %를 차지하는 투명 세포형 신장암의 발생 원인은 암 억제 유전자인 VHL(von-Hippel-Lindau) 유전자의 결손인 것으로 알려져 있다. VHL 유전자의 결손은 HIF-α/VHF의 회합 장해를 통해 저산소 유도 유전자군의 전사 활성화를 초래하여, VEGF, TGFβ 등의 유전자의 발현 증강이 발생한다고 알려져 있다.Ultrasound, CT, angiography, etc. are examined for kidney cancer, but specific biochemical markers are not known. Therefore, examination using a device is necessary. In addition, although several types of kidney cancer can be classified pathologically, the cause of the development of clear cell type kidney cancer, which accounts for about 90%, is known to be a defect in the von-Hippel-Lindau (VHL) gene, a cancer suppressor gene. . Deletion of the VHL gene results in transcriptional activation of the hypoxic inducible gene group through association of HIF-α / VHF, leading to enhanced expression of genes such as VEGF and TGFβ.
신장암의 주 치료법은 외과 요법이다. 병의 시기에 관계없이, 적출할 수 있는 경우에는 신장의 적출, 또는 신장의 부분적 적출이 가장 일반적이며, 만일 전이가 있다 하더라도 신장의 외과적 적출이 고려되는 경우가 있다. 외과 요법 이외의 방법으로서는 신동맥의 동맥색전술이 있으며, 이 방법은 적출이 불가능한 경우나, 큰 종양을 적출하는 경우에, 수술에 앞서서 시행되는 경우가 있다. 최근 화상 진단 기술의 발전에 의해 신장암은 매우 작은 크기여도 조기에 발견되도록 되어, 초기 암에서의 치료에서는 90% 이상이 치유된다. 그러나, 5 ㎝ 이상의 큰 종양이나 전이가 있는 종양의 치료 성과는 빈약하기 때문에(신장 세포암 전체의 5년 생존율은 약 50 내지 60%), 조기 발견의 중요성이 인식되고 있다.
The main treatment for kidney cancer is surgical therapy. Regardless of the timing of the disease, extraction of the kidneys or partial extraction of the kidneys is the most common where possible, and even if there is metastasis, surgical removal of the kidneys may be considered. Arterial embolization of the renal artery may be used as a method other than the surgical therapy. This method may be performed prior to surgery in cases where it is impossible to extract or when a large tumor is removed. Recent advances in burn diagnostics have led to early detection of renal cancer, even at very small sizes, resulting in more than 90% healing in early cancer. However, since the therapeutic results of large tumors larger than 5 cm or tumors with metastases are poor (the 5-year survival rate of all renal cell carcinomas is about 50 to 60%), the importance of early detection is recognized.
한편, 마자인(Cannabis Semen)은 뽕나무과(Moraceae)의 삼(Cannabis sativa L.)의 종자를 건조한 것으로서, 그 기원식물인 삼은 중앙아시아 원산의 일년초로서 원줄기는 높이 1-2.5 m로서 곧추 자라며 둔한 사각형이고 잔털이 있으며 녹색이다. 잎은 밑부분에서는 대생하고 엽병이 길며 5-9개로 갈라진 장상복엽이지만, 윗부분에서는 3개로 갈라지거나 갈라지지 않고 엽병도 짧다. 삼의 성분으로 알려진 테트라하이드로카나비놀(tetrahydrocannabinol)은 향정신성 작용을 가지며, 중추신경, 대뇌 척수에 대하여 처음에는 쾌락, 환각을 동반하는 흥분이 오는 데, 이것이 경과되면 마비작용으로 이행된다. 동시에 감각령에 둔마가 와서 통각의 소실을 가져오고, 결국 동공산대에 이른다. 마자인은 장을 윤택하게 하며, 행인, 작약, 지실, 대황, 후박과 배합한 마자인환은 변비를 치료하는 데 사용한다. 즉 마자인은 장을 통하게 하여 변이 잘 나오게 한다. 변비에 응용할 수 있다(박종희, 이정규 공저, 상용 약용식물도감, p199-p201). 마자인은 마른 것을 윤택하게 하고, 장을 부드럽게 소통하도록 도와주며 혈액순환을 순조롭게 하는 효능이 있는 약재이다.On the other hand, Cannabis Semen is a dried seed of Cannabis sativa L. of the genus Moraceae, whose origin is the annual herb of Central Asia, whose main stem is 1-2.5 m high and grows straight. With fine hairs and green. The leaves are vivid at the bottom, long lobes, long intestinal bile divided into 5-9, but the upper part is shorted without splitting or splitting into three. Tetrahydrocannabinol, known as hemp, has psychotropic effects and initially produces pleasure and hallucination with the central nervous system and cerebral spinal cord, which pass into paralysis. At the same time, the dense horse comes to the sensory spirits, bringing about the loss of pain, and eventually reaches the communist zone. As soon as it is intestine moisturizes, and combined with passersby, peony, fruit room, rhubarb, bakbak is used to treat constipation. In other words, as soon as the chapter through the stools to come out well. It can be applied to constipation (Park Jong-hee, co-author Lee Jeong-gyu, commercial medicinal plant illustration, p199-p201). As soon as it is dry, it helps to smoothly communicate the intestines and improves blood circulation.
즉, 마자인(Cannabis Semen)은 삼(cannabis sativa L.)(뽕나무과 Moraceae)의 씨로서 한방과 민간에서 윤장통변, 혈허진휴, 장조변비, 위장조결, 비약변비 등의 목적으로 사용하였다.
In other words, Cannabis Semen ( Cannabis Semen ) is a seed of hemp ( cannabis sativa L.) (Moraceae family) used for the purpose of yunjangcheong, hemophilia, intestinal constipation, gastrointestinal tightening, medicinal constipation in oriental medicine and private.
대한민국 공개특허공보 제2003-0059951호에는 마자인을 포함하는 생약 복합제 즉, 대황, 차전자, 욱리인, 빈랑, 마자인, 토사자, 우슬, 산약, 산수유, 지실, 방풍 및 독활로 이루어진 추출물을 포함하는 당뇨병 예방 및 치료용 조성물이 개시되어 있고, 대한민국 공개특허공보 특1999-0084271호에는 마자인을 포함하는 변비증상 개선 효과가 있는 차 및 이의 제조방법이 개시되어 있으며, 대한민국 등록특허공보 제10-0630410호에는 마자인 추출물을 함유하는 여드름 예방 및 치료를 위한 외용제 조성물이 개시되어 있다. 이 발명은 낮은 농도에서 여드름균(P. acnes)의 증식억제 효과를 보이는 귀판, 노회, 마자인, 백굴채 및 현호색의 효과를 확인하고 이러한 증식 억제능을 가진 생약추출물을 함유하는 조성물에 대한 것이다. 그러나, 마자인의 신장암 관련 질환과 관련하여 상기 문헌 어디에도 개시되거나 공개된 바가 없다.
Korean Unexamined Patent Publication No. 2003-0059951 includes a herbal compound containing mazain, that is, an extract consisting of rhubarb, chason, ulriin, betel nut, mazain, earth and sand, dew, pesticide, cornus, fruiting, windproof and poisonous A composition for preventing and treating diabetes is disclosed, and Korean Laid-Open Patent Publication No. 1999-0084271 discloses a tea having a constipation symptom improvement effect including mazain and a method for manufacturing the same, and the Republic of Korea Patent Publication No. 10- 0630410 discloses a topical composition for the prevention and treatment of acne containing the extract of Mazarin. The present invention relates to a composition containing the herbal extract having the ability to inhibit the growth of the ear, synagogues, maine, white oysters and corydalis showing a proliferation inhibitory effect of P. acnes at low concentrations. However, nothing has been disclosed or disclosed in this document in connection with kidney cancer-related diseases of mazain.
이에, 본 발명자들은 마자인에 대한 생약 연구를 하던 중, 마자인 추출물이 신장암 세포를 효과적으로 사멸시킬 수 있음을 확인함으로써 본 발명을 완성하였다.
Thus, the present inventors completed the present invention by confirming that the extract of the mother can effectively kill kidney cancer cells during the study of herbal medicine for the mother.
본 발명의 목적은 마자인 추출물을 유효성분으로 함유하는 신장암 치료용 조성물 및 기능성 식품을 제공하는 것이다.
It is an object of the present invention to provide a composition for treating kidney cancer and a functional food containing the extract as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 마자인(Cannabis Semen) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방 및 치료용 조성물을 제공한다.In order to achieve the above object, the present invention provides a composition for preventing and treating kidney cancer containing Cannabis Semen ethanol extract as an active ingredient.
또한, 본 발명은 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 마자인(Cannabis Semen) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방용 기능성 식품을 제공한다.
In addition, the present invention provides a functional food for preventing kidney cancer containing Cannabis Semen ethanol extract containing a food supplement acceptable food supplement as an active ingredient.
이하, 본 발명을 상세히 설명한다. Hereinafter, the present invention will be described in detail.
본 발명은 마자인(Cannabis Semen) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방 및 치료용 조성물을 제공한다. 본 발명의 조성물은 활성성분으로서 마자인 추출물을 포함하며, 추가적으로 약제학적으로 허용가능한 담체 또는 희석제를 포함할 수 있다.The present invention provides a composition for the prevention and treatment of kidney cancer containing Cannabis Semen ethanol extract as an active ingredient. The composition of the present invention comprises a extract as soon as the active ingredient, and may further comprise a pharmaceutically acceptable carrier or diluent.
본 발명의 신장암 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 kidney cancer of the present invention, the ethanol extract is preferably extracted at 50 ° C. for 24 hours, wherein the ethanol extract is more preferably dried and concentrated at 45 ° C. under reduced pressure, and the ethanol Is most preferably 95%.
또한, 본 발명의 신장암 예방 및 치료용 조성물에 있어서, 상기 신장암은 신세포암(kindney cell cancer)인 것이 바람직하다.In addition, in the composition for preventing and treating kidney cancer of the present invention, the kidney cancer is preferably renal cell cancer.
본 발명의 마자인 추출물은 마자인의 어느 부위에서도 가능하지만, 바람직하게는 뿌리로부터 추출되는 것이 바람직하다. 그리고 추출용액은 물 또는 유기용매로 추출하여 얻을 수 있는데, 유기용매로는 저급 알콜, 아세톤, 클로로포름, 메틸렌클로라이드, 에테르, 에틸아세테이트, 헥산 등을 예시할 수 있다. 저급알콜로는 메탄올, 에탄올, 프로판올 및 부탄올을 예시할 수 있으며, 에탄올이 가장 바람직하다.As soon as the extract of the present invention is possible at any site of the mother, it is preferably extracted from the root. The extraction solution may be obtained by extraction with water or an organic solvent, and examples of the organic solvent may include lower alcohols, acetone, chloroform, methylene chloride, ether, ethyl acetate, and hexane. Lower alcohols include methanol, ethanol, propanol and butanol, with ethanol being most preferred.
구체적으로, 마자인 건조물 또는 분말에 1 내지 5 배, 바람직하게는 3배의 95% 에탄올을 첨가하고 20 내지 100℃, 바람직하게는 40 내지 60℃의 온도에서 10 내지 100시간, 바람직하게는 15 내지 40시간, 더욱 바람직하게는 24시간 동안 추출한 후 여과하여 마자인의 에탄올 추출물을 제조할 수 있다. 바람직하게는 상기 추출액을 여과하여 얻어진 여액을 감압농축하여 제조할 수 있다. 상기 추출방법들에서 추출 공정은 필요에 따라 2회 이상 반복하여 실시할 수 있으며, 여과 후 얻어진 추출물을 동결 건조 또는 감압건조시켜 분말 형태로 만들 수도 있다.
Specifically, 1 to 5 times, preferably 3 times, 95% ethanol is added to the dried or powder as soon as 10 to 100 hours, preferably 15 at a temperature of 20 to 100 ° C, preferably 40 to 60 ° C. After extracting for 40 hours, more preferably 24 hours, the ethanol extract may be prepared by filtering. Preferably, the filtrate obtained by filtering the extract may be concentrated under reduced pressure. In the above extraction methods, the extraction process may be repeated two or more times as necessary, and the extract obtained after filtration may be lyophilized or dried under reduced pressure to obtain a powder form.
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 "pharmaceutically acceptable carrier" is a pharmaceutically acceptable substance such as a liquid or solid filler, diluent, excipient or solvent which serves to transport the active ingredient from one organ or part of the body to another organ or part of the body. , Composition or vehicle.
본 발명의 신장암 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 kidney cancer of the present invention may be prepared as a medicament by adding one or more pharmaceutically acceptable carriers together with the active ingredient. The carrier may include, but is not limited to, saline, buffered saline, water, glycerol and ethanol, and any suitable agent known in the art (Remingtons's Pharmaceutical Science (Recent Edition), Mack Publishing Company, Easton PA) may be used. .
본 발명의 마자인 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.Formulations for the formulation of the extract of the present invention can be administered orally at the time of clinical administration and can be used in the form of general pharmaceutical formulations, when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants And diluents such as surfactants or excipients. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, etc., and liquid preparations for oral use include suspensions, solvents, emulsions, and syrups. In addition to liquids and paraffins, various excipients may be included, such as wetting sweeteners, fragrances, preservatives and the like.
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 that may be added to the composition of the present invention may be any pharmaceutically acceptable herbal medicine, for example, Angelica tenuissimae Radix, Gastrodiae Rhizoma, Bapleuri Radix, Angelica ( Angelicae gigantis Radix, Persicae Semen, Cinnamomi Ramulus, Rhubarb (Rhei Rhizoma), Licorice (Glycyrrhizae Radix), Cnidii Rhizoma, Aurantii nobilis Pericarpium, Taxa (Alismatis Rhizoma) Coptidis Rhizoma, Scutellariae Radix, Hoelen, Peeoniae Radix, Atractylodis Rhizoma alba, Phellodendri Cortex, Gardeniae Fructus, Pinelliae Tuber, Ramulu Set (Uncaria) Uncus, Ponciri Fructus, Ginseng (Gingseng), Liriopis Tuber, Polygalae Radix, Acori graminei Rhizoma, Atractylodis Rhizoma alba, Chrysanthemi Flos, Windproof (Ledebouri) ), Ginger (Zingiberis Rhizoma crudus), forget-me-not (Natrii sulfas), control (Ziz) yphi Fructus, Salviae Radix, Mautan Radicis Cortex, Rehmanniae Radix, Mint Herba, Dioscoreae Rhizoma, Polyporus, Polygoni multiflori Radix, Gusi (Allii tube) Semen, Cassiae Semen, Lycii Fructus, Araliae cordatae Radix, Eucommiae Cortex, Hedyotis Herba, Saururus Herba, Artemisiaecapillaris Herba, Anemarrhen Rhizoma, Carthami Flos, Astragali Radix, Lycopodium, Ginkgonis Folium, Polygonati Rhizoma, Nelumbinis Semen, Fossilia ossis Mastodi, Cortexi radics ), Achyranthis Radix, Rehmanniae Radix preparata, Perillae Semen, Thujae Semen, Malt (Hordei Fructus germinatus), Cuscutae Semen, Mordaedae Radix, Sea Song (Pini koraiensis) Radix) and the like can 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 may be administered in various parenteral formulations during actual clinical administration, and solid preparations include tablets, pills, powders, granules, capsules, and the like. In addition to water, liquid, and paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. Specifically, preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, 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. In addition, calcium or vitamin D 3 may be added to enhance the efficacy of the treatment. Such compositions may be presented in unit-dose (single) or multi-dose (several) containers, such as sealed ampoules and vials, and immediately before use, sterile liquid carriers such as injectable water. Can be stored under freeze-drying conditions requiring only the addition of. Immediate injection solutions and suspensions can 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 can be applied differently depending on the age, sex, condition of the subject, the absorption of the active ingredient in the body, the inactivation rate and excretion rate, the drug used in combination. The invention also includes formulations of dosage units. The formulations are present in individual dosage forms, such as tablets, coated tablets, capsules, pills, suppositories, and ampoules, wherein the amount of active compound in the drug corresponds to the fraction or multiple of the individual dosage. Dosage units may contain, for example, one, two, three or four times the individual dosage, or 1/2, 1/3 or 1/4 times. The individual dosages preferably contain an amount in which the active compound is administered at one time, which usually corresponds to all, 1/2, 1/3 or 1/4 times the daily dosage.
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" refers to an active ingredient isolated from natural products. The extract may 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 any form formulated using the same.
본 발명의 구체적인 실시에서, 마자인의 에탄올 추출물은 ACHN 신장암 세포를 100 ㎍/㎖에서 89.9% 사멸시키고, EC50(half maximal effective concentration)은 39.7 ㎍/㎖이었다. 상기 결과는 본 발명의 마자인 추출물이 우수한 ACHN 신장암 세포의 사멸 활성을 가지며, 나아가 신장암 치료 및 예방 활성을 가진다는 것을 입증하는 것이다.
In a specific embodiment of the present invention, the ethanol extract of mazain killed 89.9% of ACHN kidney cancer cells at 100 μg / ml and the EC 50 (half maximal effective concentration) was 39.7 μg / ml. The above results demonstrate that the extract of the present invention has excellent killing activity of ACHN renal cancer cells, and further has renal cancer treatment and prophylactic activity.
본 발명에서 용어, "예방"이란 조성물의 투여로 신장암 발병을 억제 또는 지연시키는 모든 행위를 의미한다.As used herein, the term "prevention" means any action that inhibits or delays the development of kidney cancer by administration of the composition.
본 발명에서 용어, "치료"란 조성물의 투여로 신장암의 증세가 호전되거나 이롭게 변경하는 모든 행위를 의미한다.As used herein, the term "treatment" means any action that improves or advantageously changes the symptoms of kidney cancer by administration of the composition.
본 발명에서 마자인 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.As soon as the extract in the present invention can be extracted using water, an organic solvent, or a mixed solvent thereof. Preferably it is extracted using an organic solvent, in particular ethanol. The extracted solution can be used directly or can be concentrated and / or dried. When extracted with an organic solvent, methanol, ethanol, isopropanol, butanol, ethylene, acetone, hexane, ether, chloroform, ethyl acetate, butyl acetate, dichloromethane, N, N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO), 1,3-butylene glycol, propylene glycol, or a mixed solvent thereof may be used and extracted by room temperature or warming under conditions where the active ingredient of the herbal medicine is not destroyed or minimized. Depending on the organic solvent to be extracted, the degree of extraction and loss of the active ingredient of the drug may vary, so select an appropriate organic solvent. The extraction method is not particularly limited, and examples thereof include cold needle extraction, ultrasonic extraction, reflux cooling extraction, and the like. Filtration is a process of removing the suspended solid particles from the extract, it may be used to filter the particles using cotton, nylon or the like, or may be used, such as ultrafiltration, cryofiltration, centrifugal separation, but is not limited thereto.
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 may be used, such as concentrated under reduced pressure, reverse osmosis concentration. The drying step after concentration includes freeze drying, vacuum drying, hot air drying, spray drying, reduced pressure drying, foam drying, high frequency drying, infrared drying, and the like. If desired, a process of grinding 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 can perform an additional fractionation process. Preferably, the extract is suspended in distilled water to obtain a nonpolar solvent soluble layer by extraction and separation with a nonpolar organic solvent such as hexane, ether, dichloromethane, chloroform, ethyl acetate, or a mixed solvent thereof. It can be used by concentrating and / or drying it.
본 발명에서, 용어 "약제학적으로 허용가능한 염"이란 약리학적 또는 생리학적으로 허용되는 무기산, 유기산 및 염기로부터 유도된 염을 의미한다. 적합한 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 salts" means salts derived from pharmacologically or physiologically acceptable inorganic acids, organic acids and bases. Examples of suitable acids include hydrochloric acid, bromic acid, sulfuric acid, nitric acid, perchloric acid, fumaric acid, maleic acid, phosphoric acid, glycolic acid, lactic acid, salicylic acid, succinic acid, toluene-p-sulfonic acid, tartaric acid, acetic acid, citric acid, methanesulfonic acid, 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 신장암 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 신장암에 대한 치료 활성을 가지는 물질을 포함할 수 있다.
The pharmaceutical composition for preventing and treating kidney cancer diseases of the present invention comprises 0.1 to 50% by weight of the extract or compound based on the total weight of the composition. In addition, the composition does not increase the efficacy, but may include additional ingredients that are commonly used in the pharmaceutical composition to improve the smell, taste, time and the like. In addition, the composition adds inorganic and organic additives such as vitamins B1, B2, B6, C, E, niacin, carnitine, betaine, folate pantothenic acid, biotin, zinc, iron, calcium, chromium, magnesium, and mixtures thereof. It can be included as. In addition, the composition may include a substance having a therapeutic activity against kidney cancer used alone or used in the past.
본 발명에서 용어, "환자"는 신장암 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 마자인 추출물을 포함하는 조성물을 환자에게 투여함으로써, 상기에서 언급한 신장암을 효과적으로 예방 및 치료할 수 있다. 본 발명의 조성물을 기존의 신장암 치료제와 병행하여 투여할 수 있다.As used herein, the term "patient" refers to humans, horses, sheep, pigs, goats, and camels having a disease caused by kidney cancer and its direct and indirect causes, and whose symptoms may be improved by administering the composition of the present invention. Means animals such as nutrition, dogs. By administering to a patient a composition comprising the extract of as soon as the present invention, it is possible to effectively prevent and treat the above-mentioned kidney cancer. The composition of the present invention can be administered in parallel with existing kidney cancer treatments.
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.As used herein, the term "administration" means introducing a predetermined substance into a patient by any suitable method, and the route of administration of the composition of the present invention is oral or parenteral via any general route as long as the target tissue can be reached. May be administered. In addition, the composition may be administered by any device in which the active agent may migrate 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. As used herein, the term “pharmaceutically effective amount” means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and an effective dose level refers to a patient's sexually transmitted disease, age, severity, and drug activity. , Drug sensitivity, time of administration, route of administration and rate of release, duration of treatment, factors including concurrently used drugs, and other factors well known in the medical arts. The compositions of the present invention may be administered as individual therapeutic agents or in combination with other therapeutic agents and may be administered sequentially or simultaneously with conventional therapeutic agents. It may be single or multiple doses. 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 of administering the composition comprising the extract or compound prepared according to the preparation method of the present invention is preferably oral administration or intravenous administration. In general, when the effective dose is oral administration, it is usually 1 to 1 time per adult. 500 mg / kg is preferred, and in the case of intravenous administration, 1 to 100 mg / kg is preferred, and may be administered 2-3 times a day. Dosage levels for a particular patient may vary depending on sex, age, health condition, diet, time of administration, method of administration, drug mixture, the condition of the patient, and the extent of the onset of neurological disease.
또한, 본 발명은 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 마자인(Cannabis Semen) 에탄올 추출물을 유효성분으로 함유하는 신장암 예방용 기능성 식품을 제공한다.In another aspect, the present invention provides a functional food for preventing kidney cancer containing Cannabis Semen ethanol extract containing a food supplement acceptable food 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.Functional food of the present invention preferably contains 0.1 to 5% by weight, more preferably 1% by weight of the ethanol extract relative to the total weight. Functional food of the present invention is not particularly limited in the formulation, for example, dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, Various soups, beverages, tea, drinks, alcoholic beverages and vitamin complexes, etc., and includes all of the health food in the usual 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, "functional food" means a food manufactured and processed using raw materials or ingredients having functional properties useful for the human body according to Act No. 6767 of the Health Functional Food Act, and "functional" refers to the structure of the human body And ingestion for the purpose of obtaining nutrients for function or for obtaining useful effects in health uses such as physiological actions.
본 발명에서는 ACHN 신장암 세포에 본 발명의 마자인 추출물을 투입한 결과, ACHN 신장암 세포가 사멸하는 것을 확인하였다(도 1 참조). 따라서, 마자인 추출물이 신장암 치료 효과가 있다는 것을 새롭게 알 수 있다. 이에, 본 발명에서는 마자인 추출물을 유효성분으로 함유하는 신장암 예방용 기능성 식품을 제조함으로써 본 발명을 완성하였다(제조예 2 참조).
In the present invention, as a result of injecting the extract of the present invention into ACHN kidney cancer cells, it was confirmed that the ACHN kidney cancer cells die (see Fig. 1 ). Therefore, it can be seen that the extract as soon as there is a therapeutic effect for kidney cancer. Thus, in the present invention, the present invention was completed by preparing a functional food for preventing kidney cancer containing the extract as soon as the active ingredient (see Preparation Example 2 ).
상기에서 살펴본 바와 같이, 본 발명의 마자인 추출물은 신장암 세포의 성장을 억제하고 세포사멸을 유도한다. 따라서 본 발명에 따른 신장암 치료용 조성물은 신장암 환자의 치료에 매우 효과적일 것이다.
As described above, as soon as the extract of the present invention inhibits the growth of kidney cancer cells and induces apoptosis. Therefore, the composition for treating renal cancer according to the present invention will be very effective in the treatment of renal cancer patients.
도 1은 인간 신장암 ACHN 세포에서 마자인의 도입이 신장암 세포의 성장에 미치는 영향을 알아보기 위한 Alamar Blue 분석 결과이다. 도면의 가로축은 추출물의 농도를, 세로축은 신장암 세포의 생존률을 나타낸다.
1 is a result of the Alamar Blue analysis to determine the effect of the introduction of Mazarin in human kidney cancer ACHN cells on the growth of kidney cancer cells. In the figure, the horizontal axis shows the concentration of the extract, and the vertical axis shows the survival rate of kidney 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> 마자인 추출물의 제조Example 1 Preparation of Mazarine Extracts
서울 약재상에서 구입한 마자인(중국산) 3 ㎏을 실온에서 5일간 건조하고 분쇄하였다. 상기 분쇄된 마자인을 95% 에탄올(ethanol) 30 ℓ에 침지시키고 50℃에서 24시간 동안 추출하였다. 이것을 여과지를 통하여 여과한 후 45℃ 감압 조건에서 건조 및 농축하여 총 추출물 418 g을 수득하고, -20℃에서 보관하였다.
3 kg of mazain (made in China) purchased from Seoul medicinal herb was dried and ground at room temperature for 5 days. The ground mazain was soaked in 30 L of 95% ethanol and extracted at 50 ° C. for 24 hours. This was filtered through filter paper, dried and concentrated under reduced pressure at 45 ° C. to obtain 418 g of the total extract, which was stored at −20 ° C.
<실시예 2> 마자인 추출물이 신장암 세포의 성장에 미치는 영향Example 2 Effect of Mazain Extract on the Growth of Kidney Cancer Cells
상기 실시예 1에서 추출한 마자인 추출물이 신장암 세포의 성장에 미치는 영향을 알아보기 위하여, 인간 신장암 세포인 ACHN 세포에 대하여 마자인의 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.
In order to determine the effect of the extract extracted from Example 1 on the growth of renal cancer cells, ACHN cells, which are human renal cancer cells, were treated with ethanol extracts of madin for 48 hours and subjected to Alamar Blue analysis. The Alamar Blue assay is a modified form of the MTT assay, in which a specific enzyme degrades a living cell and then measures the fluorescence intensity of the product as the compound breaks down to determine the relative number of living cells after treatment. I am an experimental method. It will be described in more detail below.
<2-1> 인간 신장암 세포주의 준비 및 처리<2-1> Preparation and Treatment of Human Kidney Cancer Cell Line
본 발명에 사용된 신장암 세포주인 ACHN 세포는 한국세포주은행(Korean Cell Line Bank, KCLB)으로부터 분양받아 실험에 이용하였다. 구체적으로, ACHN 신장암 세포를 10% FBS(fetal bovine serum, 소태아혈청)(Welgene)와 25 mM HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid)를 포함하는 MEM(Minimum essential medium) 배지에서 계대배양하였다.
ACHN cells, a kidney cancer cell line used in the present invention, were distributed from Korean Cell Line Bank (KCLB) and used for experiments. Specifically, ACHN kidney cancer cells (Minimum essential medium) containing 10% FBS (fetal bovine serum, fetal bovine serum) (Welgene) and 25 mM HEPES (4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid) Subcultured in medium.
<2-2> ACHN 신장암 세포의 세포 성장 억제 측정<2-2> Cell growth inhibition measurement of ACHN kidney cancer cells
상기 실시예 1에서 추출한 마자인 추출물이 신장암 세포인 ACHN 세포에 대하여 성장을 억제시키는 효과를 확인하였다. 구체적으로, 96 웰 플레이트에 각 웰 당 6.0× 103 개의 ACHN 신장암 세포를 주입(seeding)하고 24시간 동안 배양한 후, DMSO(Dimethyl sulfoxide)에 녹인 상기 마자인 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 및 도 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, 신장암 세포의 생존율을 도 1에 나타내었다.
As soon as the extract extracted in Example 1 was confirmed the effect of inhibiting growth against ACHN cells which are kidney cancer cells. Specifically, after injection of 6.0 × 10 3 ACHN kidney cancer cells per well into a 96 well plate and incubating for 24 hours, the mother dissolve in DMSO (dimethyl sulfoxide) When the ethanol extract was treated at concentrations of 0 to 100 μg / ml (specifically, concentrations of 0, 3.125, 6.25, 12.5, 25, 50, and 100 μg / ml, respectively) for 48 hours, the degree of inhibition of cell growth was confirmed. (Table 1 and FIG. 1). After treatment with each concentration of extract, 20 μl of Alamar Blue reagent was added to 0.2 ml of cell culture filled in each well in a 96-well plate, and the plates were incubated for 2 hours in an incubator. The plate was slowly shaken to evenly react the cells in each well, and the intensity of fluorescence was measured at 590 nm with a Fluorescence Microplate Reader (Molecular Devices Corp.) while irradiating irradiated light at a wavelength of 544 nm. The survival rate of is shown in FIG.
그 결과, 표 1 및 도 1에서 나타난 바와 같이, 마자인의 처리 농도가 높을수록 신장암 세포의 성장이 감소하였으며, 이로부터 마자인이 신장암 치료 효과를 가짐을 알 수 있었다. 즉, 3.125 ㎍/㎖에서 2.0%, 6.25 ㎍/㎖에서 3.3%, 12.5 ㎍/㎖에서 8.3%, 25 ㎍/㎖에서 26.2%, 50 ㎍/㎖에서 61.7%, 100 ㎍/㎖에서 89.9%로 신장암 세포를 사멸시켰다. 아울러, EC50(half maximal effective concentration)은 39.7 ㎍/㎖로 측정되었다. 상기 표 1에서 마자인을 처리하지 않은 대조군의 신장암 세포의 생존율 수를 1을 기준으로 하여 각각의 마자인 처리 농도에 따른 48시간 후의 신장암 세포의 상대적 세포수를 기재하였다. 이와 같이, 본 발명의 마자인 추출물은 우수한 ACHN 세포 사멸 활성을 가지며, 나아가 신장암 치료 및 예방 활성을 가진다는 것을 입증한다.
As a result, as shown in Table 1 and Figure 1, as the concentration of the treatment of the higher the growth of kidney cancer cells was reduced, it can be seen that as soon as having the effect of renal cancer treatment. Ie 2.0% at 3.125 μg / ml, 3.3% at 6.25 μg / ml, 8.3% at 12.5 μg / ml, 26.2% at 25 μg / ml, 61.7% at 50 μg / ml, 89.9% at 100 μg / ml Kidney cancer cells were killed. In addition, an EC 50 (half maximal effective concentration) was determined to be 39.7 μg / ml. In Table 1, the relative cell numbers of kidney cancer cells after 48 hours according to the concentrations of the respective treatments based on the number of survival rates of the kidney cancer cells of the control group not treated with mazain were described as 1. As such, the extract of the present invention has excellent ACHN cell killing activity and further demonstrates that it has therapeutic and prophylactic activity for kidney cancer.
<실시예 3> 마자인 추출물에 의한 급성독성 시험Example 3 Acute Toxicity Test with Mazarin Extracts
본 발명에 이용된 마자인은 널리 약재로 이용되고 있어서 안정성에 문제가 없을 것으로 판단하였으나, 경구 투여시 및 복강내 투여시의 독성 실험을 수행하여 이를 확인하고자 하였다.As soon as it was determined that there is no problem in stability as it is widely used as a medicinal herb, the purpose of this study was to confirm the toxicity by oral administration and intraperitoneal administration.
6주령의 특정병원부재(SPF) SD계 랫트를 사용하여 급성독성실험을 실시하였다. 군당 2 마리씩의 동물에 본 발명의 실시예 1의 마자인 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 test was performed using 6-week-old SPF SD rats. Two animals per group were suspended orally administered at a dose of 5 g / kg, each of the extract of Example 1 of the present invention suspended in 0.5% methylcellulose solution. After administration of the test substance, mortality, clinical symptoms, and changes in body weight were observed, and hematological and hematological examinations were performed.
시험결과, 시험물질을 투여한 모든 동물에서 특기할 만한 임상증상이나 폐사된 동물은 없었으며, 체중변화, 혈액검사, 혈액생화학 검사, 부검소견 등에서도 독성변화는 관찰되지 않았다. 이상의 결과 마자인 추출물은 모두 랫트에서 5 g/㎏까지 독성변화를 나타내지 않으며 경구 투여 최소치사량 (LD50)은 5 g/㎏이상인 안전한 물질로 판단되었다.
As a result, there were no clinical symptoms or deaths in all animals treated with the test substance, and no toxicity change was observed in weight change, blood test, blood biochemistry test, autopsy findings, etc. As a result, all of the extracts did not show toxic changes up to 5 g / kg in rats, and the minimum lethal dose (LD 50 ) was determined to be a safe substance of 5 g / kg or more.
<제조예 1> 마자인 추출물을 유효성분으로 함유하는 신장암 치료제의 제조Preparation Example 1 Preparation of a Renal Cancer Therapeutic Agent Containing Mazarin Extract as an Active Ingredient
본 발명자들은 상기 실시예를 통해 마자인 추출물의 신장암 치료 효능이 뛰어남을 확인하여 마자인 추출물을 유효성분으로 함유하는 신장암 치료제를 하기와 같이 제조하였다. 또한, 하기 치료제의 제조예는 치료제 뿐만 아니라 건강식품의 제조에도 응용하여 사용될 수 있다.
The present inventors have confirmed that the kidney cancer treatment effect of the extract as soon as through the above example was prepared for the treatment of kidney cancer containing the extract as an active ingredient as soon as. In addition, the preparation of the following therapeutic agent can be used for the application of not only a therapeutic agent but also a health food.
<1-1> 마자인 추출물을 함유하는 연질캅셀(soft gelatin capsules)<1-1> Soft gelatin capsules containing Mazarin extract
마자인 추출물 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%10% wax
팜유 25%Palm oil 25%
홍화씨유 30.399%
Safflower Seed Oil 30.399%
<1-2> <1-2> 마자인Mazain 추출물을 함유하는 정맥주사용 제제의 제조 Preparation of Intravenous Formulation Containing Extract
마자인 추출물 0.2%Mazarin Extract 0.2%
만니톨 0.3%Mannitol 0.3%
생리식염수 9.5%
Saline 9.5%
<1-3> 마자인 추출물을 함유하는 정제(tablet)<1-3> Tablet containing Mazarin extract
마자인 추출물 35%Mazarin 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> 마자인 추출물을 유효성분으로 함유하는 기능성 식품의 제조Preparation Example 2 Preparation of Functional Foods Containing Mazain Extract as Active Ingredient
본 발명자들은 상기 실시예를 통해 마자인 추출물이 신장암 치료 활성이 뛰어남을 확인하여 이를 유효성분으로 함유하는 기능성 식품을 하기와 같이 제조하였다.
The present inventors confirmed that as soon as the extract extract as soon as the kidney cancer therapeutic activity through the above-mentioned to prepare a functional food containing it as an active ingredient as follows.
<2-1> 음료의 제조<2-1> Preparation of Drink
꿀 522 ㎎522 mg of honey
치옥토산아미드 5 ㎎Chioctosanamide 5 mg
니코틴산아미드 10 ㎎Nicotinamide 10 mg
염산리보플라빈나트륨 3 ㎎Riboflavin Sodium Hydrochloride 3 mg
염산피리독신 2 ㎎Pyridoxine hydrochloride 2 mg
이노시톨 30 ㎎Inositol 30 mg
오르트산 50 ㎎Orthoic acid 50 mg
마자인 추출물 0.48 ~ 1.28 ㎎Mazarin Extract 0.48 ~ 1.28 mg
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%
마자인 추출물 0.24 ~ 0.64 %Mazarin Extract 0.24 ~ 0.64%
후르츠향 1 %1% fruit flavor
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> Preparation of Candy
설탕 50 ~ 60 %Sugar 50-60%
물엿 39.26 ~ 49.66 %Starch syrup 39.26 ~ 49.66%
마자인 추출물 0.24 ~ 0.64 %Mazarin Extract 0.24 ~ 0.64%
오렌지향 0.1 %Orange flavor 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> Preparation of Biscuits
박력1급 88 ㎏Force Class 1 88 kg
중력1급 76.4 ㎏Gravity First Class 76.4 ㎏
정백당 16.5 ㎏16.5 kg
식염 2.5 ㎏2.5 kg of salt
포도당 2.7 ㎏2.7 kg of glucose
팜쇼트닝 40.5 ㎏Palm shortening 40.5 kg
암모 5.3 ㎏5.3 kg of ammo
중조 0.6 ㎏Medium kg 0.6 kg
중아황산나트륨 0.55 ㎏0.55 kg sodium bisulfite
쌀가루 5.0 ㎏Rice flour 5.0 kg
비타민 B1 0.003 ㎏Vitamin B1 0.003 kg
비타민 B2 0.003 ㎏0.003 kg of vitamin B2
밀크향 0.16 ㎏Milk Flavor 0.16 ㎏
물 71.1 ㎏71.1 kg of water
전지분유 4 ㎏Whole milk powder 4 kg
대용분유 1 ㎏Substitute powder 1 kg
제일인산칼슘 0.1 ㎏0.1 kg of calcium phosphate
살포염 1 ㎏Spray salt 1 kg
분무유 25 ㎏25 kg of spray oil
마자인 추출물 0.2 ~ 0.5 ㎏Mazarin Extract 0.2 ~ 0.5 kg
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 비스켓을 제조하였다.
Biscuits were prepared using conventional methods with the above composition and content.
<2-5> 아이스크림의 제조<2-5> Preparation of Ice Cream
유지방 10.0 %Milkfat 10.0%
무지유고형분 10.8 %Non-fat solids 10.8%
설탕 12.0 %Sugar 12.0%
물엿 3.0 %Starch syrup 3.0%
유화안정제(스팬, span) 0.5 %Emulsifying stabilizer (span) 0.5%
향료(스트로베리) 0.15 %Spices (Strawberries) 0.15%
물 63.31 ~ 62.91 %Water 63.31 ~ 62.91%
마자인 추출물 0.24 ~ 0.64 %Mazarin Extract 0.24 ~ 0.64%
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 아이스크림을 제조하였다.
Ice cream was prepared using conventional methods using the above composition and content.
<2-6> 쵸코렛의 제조<2-6> manufacturing 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 %0.5% vanilla
마자인 추출물 0.24 ~ 0.64 %Mazarin Extract 0.24 ~ 0.64%
상기 조성 및 함량으로 하여 통상적인 방법을 사용하여 초코렛을 제조하였다.With the above composition and content, chocolate was prepared using conventional methods.
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.
On the other hand, the specific scope of the present invention is defined by the claims rather than the embodiments described above, all changes and modifications derived from the meaning and scope and equivalent concepts of the claims to the scope of the invention It should be interpreted as including.
Claims (7)
Cannabis Semen ( Cannabis Semen ) A composition for preventing and treating kidney cancer, which contains ethanol extract as an active ingredient.
According to claim 1, wherein the ethanol extract is a composition for preventing and treating kidney cancer, characterized in that extracted for 24 hours at 50 ℃.
The composition for preventing and treating kidney cancer according to claim 1 or 2, wherein the ethanol extract is dried and concentrated at 45 ° C under reduced pressure.
According to claim 1 or claim 2, wherein the ethanol is a composition for the prevention and treatment of kidney cancer, characterized in that 95%.
The method of claim 1, wherein the composition is a composition for preventing and treating kidney cancer, characterized in that it comprises a pharmaceutically acceptable carrier or diluent.
The composition for preventing and treating kidney cancer according to claim 1, wherein the kidney cancer is renal cell cancer.
Functional food for the prevention of kidney cancer containing Cannabis Semen ethanol extract containing a food supplement acceptable food supplement as an active ingredient.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020110029445A KR20120111125A (en) | 2011-03-31 | 2011-03-31 | Composition for treatment of renal cell carcinoma and functional food comprising extract of cannabis semen |
PCT/KR2012/002469 WO2012134252A2 (en) | 2011-03-31 | 2012-04-02 | Composition for renal cancer treatment and functional food containing cannabis semen extract |
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KR1020110029445A KR20120111125A (en) | 2011-03-31 | 2011-03-31 | Composition for treatment of renal cell carcinoma and functional food comprising extract of cannabis semen |
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KR20120111125A true KR20120111125A (en) | 2012-10-10 |
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KR1020110029445A KR20120111125A (en) | 2011-03-31 | 2011-03-31 | Composition for treatment of renal cell carcinoma and functional food comprising extract of cannabis semen |
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WO (1) | WO2012134252A2 (en) |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102269816B1 (en) * | 2021-01-19 | 2021-06-28 | 주식회사 네이처센스 | COMPOSITION FOR ANTI-CANCER, DECLINE IN ANTI-CANCER side effects AND Suppression OF cancer metastasis COMPRISING Extract of CANNABIS SATIVA L. |
WO2023013963A1 (en) * | 2021-08-06 | 2023-02-09 | 이재형 | Pharmaceutical composition comprising composite medicinal herb extract as an active ingredient for prevention or treatment of cancer |
KR20230022081A (en) * | 2021-08-06 | 2023-02-14 | 이재형 | Pharmaceutical composition for preventing or treating of cancer comprising medicinal herb complex extract |
Family Cites Families (4)
Publication number | Priority date | Publication date | Assignee | Title |
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KR19990070229A (en) * | 1998-02-18 | 1999-09-15 | 송시훈 | Seed Oil Manufacturing Method |
GB2418612A (en) * | 2004-10-01 | 2006-04-05 | Gw Pharma Ltd | Inhibition of tumour cell migration with cannabinoids |
JP2010520269A (en) * | 2007-03-05 | 2010-06-10 | イッサム リサーチ ディベロップメント カンパニー オブ ザ ヘブリュー ユニバーシティー オブ エルサレム | New quinonoid derivatives of cannabinoids and their use in the treatment of malignant tumors |
GB2448535A (en) * | 2007-04-19 | 2008-10-22 | Gw Pharma Ltd | New use for cannabinoid-containing plant extracts |
-
2011
- 2011-03-31 KR KR1020110029445A patent/KR20120111125A/en not_active Application Discontinuation
-
2012
- 2012-04-02 WO PCT/KR2012/002469 patent/WO2012134252A2/en active Application Filing
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR102269816B1 (en) * | 2021-01-19 | 2021-06-28 | 주식회사 네이처센스 | COMPOSITION FOR ANTI-CANCER, DECLINE IN ANTI-CANCER side effects AND Suppression OF cancer metastasis COMPRISING Extract of CANNABIS SATIVA L. |
WO2023013963A1 (en) * | 2021-08-06 | 2023-02-09 | 이재형 | Pharmaceutical composition comprising composite medicinal herb extract as an active ingredient for prevention or treatment of cancer |
KR20230022081A (en) * | 2021-08-06 | 2023-02-14 | 이재형 | Pharmaceutical composition for preventing or treating of cancer comprising medicinal herb complex extract |
Also Published As
Publication number | Publication date |
---|---|
WO2012134252A2 (en) | 2012-10-04 |
WO2012134252A3 (en) | 2013-03-07 |
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