KR101896049B1 - Composition for preventing, improving or treating prostate disease comprising extract of Hydrocotyle ramiflora as effective component - Google Patents
Composition for preventing, improving or treating prostate disease comprising extract of Hydrocotyle ramiflora as effective component Download PDFInfo
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- KR101896049B1 KR101896049B1 KR1020170061456A KR20170061456A KR101896049B1 KR 101896049 B1 KR101896049 B1 KR 101896049B1 KR 1020170061456 A KR1020170061456 A KR 1020170061456A KR 20170061456 A KR20170061456 A KR 20170061456A KR 101896049 B1 KR101896049 B1 KR 101896049B1
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- extract
- prostate
- hydrocotyle
- ramiflora
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Abstract
Description
본 발명은 큰피막이 추출물을 유효성분으로 함유하는 전립선 질환의 예방, 개선 또는 치료용 조성물에 관한 것이다.The present invention relates to a composition for preventing, ameliorating or treating a prostate disease, wherein the large capsule contains an extract as an active ingredient.
전립선은 요도가 건조해지지 않도록 여러 가지 분비물을 만들어 내는 장기로, 남성 호르몬인 안드로겐과 밀접한 관련이 있다. 분비가 증가함에 따라 점점 커지는데, 20~30대의 전립선이 50대까지 유지되다 그 이후부터 비대해진다. 이것을 양성 전립선 비대증이라고 하며, 호르몬 치료로 크기를 정상으로 돌려놓았다 하더라도 부작용의 위험이 높고, 호르몬 치료를 멈추면 곧 원상 복귀되기 때문에 완치하기 어렵다. Prostate is the organ that produces various secretions to keep the urethra from drying out. It is closely related to the male hormone, androgens. As the secretion increases, the size of the prostate increases in the 20s and 30s. This is called benign prostatic hyperplasia and it is difficult to cure it because the risk of side effects is high even if the size is returned to normal by the hormone treatment and soon after stopping the hormone treatment.
전립선에 발생하는 대표적인 질환은 전립선 비대증, 만성 전립선염 및 전립선암을 꼽을 수 있다. 전립선염은 다양한 원인이 있지만 주로 세균 감염, 원인 모를 염증, 만성통증의 일환으로 생기는 질환이고, 전립선암은 전립선 중 비교적 바깥쪽부분에 악성 종양이 생긴 것인데, 미국의 한 조사 결과에 따르면, 전립선암이 전체 남성 암 환자 중 약 35%를 차지하고 있으며, 매년 2~3%씩 증가하고 있다고 한다. 우리나라에서의 발생 빈도는 10만 명당 세 명으로 아직까지 걱정할 수위는 아니지만, 점차 서구적으로 변해가는 식생활이나 급격히 불어나는 노년층 인구로 그 비율이 점차 높아지고 있다.Representative diseases that occur in the prostate gland include enlarged prostate, chronic prostatitis, and prostate cancer. Prostate cancer is a disease caused by bacterial infections, inflammatory or chronic pain, and prostate cancer is a malignant tumor in the relatively outer part of the prostate gland. According to a US study, It accounts for about 35% of all male cancer patients and is said to increase by 2-3% every year. The incidence in Korea is three per 100,000 people, yet it is not worrisome yet, but it is gradually increasing with the western dietary life and the rapidly aging population.
한편, 전립선 비대증(Benign prostatic hyperplasia, BPH)은 노화와 성호르몬의 불균형의 영향으로 요도 주위 전립선 '샘조직'이 커져 요도를 압박하여 배뇨증상이 나타나는 질환으로, 남성의 50세 이후 발병이 급격히 일어나는 남성질환이다. 최근 우리나라에서 전립선 비대증 질환자의 증가율이 매년 20%를 상회할 정도로 가파른 증가 추세이며, 전립선이행대 부분의 평활근(smooth cell)과 상피세포(epithelial)의 과도한 증식으로 야기되고, 전립선 비대증은 하부요로증상을 동반하는데, 이는 비대해진 전립선에 의한 해부학적 폐색과 전립선 평활근을 수축시키는 α1-수용체에 의한 기능적 폐색에 기인하는 것으로 알려져 있다. 폐색에 의한 주요 증상으로는 소변줄기가 가늘고 약해지는 세뇨 또는 약뇨, 소변을 시작할 때 한참 기다려야 하는 지연뇨, 소변 중 의지와 관계없이 중단되는 배뇨중단, 소변 후 물방울처럼 떨어지는 배뇨 말기 적하, 및 잔뇨감 등이 대표적이다. 전립선 비대의 다양한 기전 중에서 대표적인 기전은 남성호르몬인 테스토스테론(testosterone,T)과 디하이드로테스토스테론(dihydrotestosterone, DHT)이 관련되어 있다고 알려져 있다.On the other hand, benign prostatic hyperplasia (BPH) is a disease in which the urethral prostate "ganglion" is enlarged due to aging and the imbalance of sex hormones, resulting in urinary symptoms due to the pressure of the urethra. It is a male disease. Recently, the rate of growth of patients with hyperplasia of the prostate in Korea has been steadily increasing to more than 20% every year. It is caused by excessive proliferation of smooth muscle and epithelial part of the prostate transplantation part, , Which is known to be due to anatomical occlusion by the enlarged prostate and functional occlusion by the? 1 -receptor which contracts the prostate smooth muscle. The main symptoms of occlusion include urinary tract weakness or weak urine stem or urine, delayed urination to wait for a long time to start urinating, discontinuation of urination to stop irrespective of urinary incontinence, terminal urination dropping like urine drops, This is representative. It is known that the testosterone (T) and the dihydrotestosterone (DHT) are involved in the typical mechanism of the various mechanisms of prostate hypertrophy.
한편, 큰피막이(Hydrocotyle ramiflora)는 산피막이풀, 큰산피막이풀이라고도 하며, 산과 들의 길가에서 자라는데, 높이가 10~15cm까지 자란다. 줄기는 옆으로 기고, 끝은 비스듬하게 서며, 가지는 곧게 서거나 비스듬히 선다. 잎은 어긋나고 둥근 모양이며 지름 1~3cm이다. 가장자리는 얕게 갈라지고 낮고 뭉툭한 톱니가 있다. 아랫쪽은 겹쳐지기도 하며, 잎자루는 길이 4~8cm이고, 턱잎은 막질(얇은 종이처럼 반투명한 것)이고, 길이 2~2.5mm이며 갈색 점이 있다. On the other hand, Hydrocotyle ramiflora grows on mountain roads and grows up to 10 ~ 15cm in height. The stem strikes to the side, the tip stood at an angle, and the stalk stands straight or at an angle. Leaves are alternate round shape with diameter of 1 ~ 3cm. The edges are shallowly divided and have low and blunt sawtooths. The petiole is 4 ~ 8cm long. The stipule is membranous (translucent like thin paper), and the length is 2 ~ 2.5mm and there are brown spots.
전립선 질환의 예방, 개선 또는 치료용 조성물로 알려진 천연 추출물에는 한국등록특허 제1653492호에 염증 또는 전립선 비대증 치료용 조성물로, 구슬꽃나무 잎 추출물 또는 이의 분획물을 유효성분으로 포함하는 것이 알려져 있으며, 일본등록특허 제5191525호에는 오람, 황 우산풍, 천심 연꽃 중에서 선택된 하나 이상의 천연물이 테스토스테론 5α-환원효소 저해제 및 안드로겐 수용체 결합 저해제로 알려져 있으나, 본 발명의 큰피막이 추출물을 유효성분으로 포함하는 전립선 질환의 예방, 개선 또는 치료용 조성물에 대해서는 개시된 바 없다. Korean Patent No. 1653492 discloses a composition for treating inflammation or enlargement of the prostate gland, which comprises a bee flower leaf extract or a fraction thereof as an active ingredient, and is known as a composition for prevention, improvement or treatment of prostate disease, Patent No. 5191525 discloses that at least one natural product selected from olam, Huang umbrella, and Tengshi lotus is known as a testosterone 5α-reductase inhibitor and an androgen receptor binding inhibitor. However, the large capsule of the present invention is useful as a preventive , Compositions for improvement or treatment have not been disclosed.
본 발명은 상기와 같은 요구에 의해 도출된 것으로서, 본 발명은 큰피막이 추출물을 유효성분으로 포함하는 전립선 질환의 예방, 개선 또는 치료용 조성물을 제공하고, 본 발명의 유효성분인 큰피막이 추출물이 전립선 비대(BPH)에 의해 비후화된 전립선의 무게를 감소시키며, 안드로겐 수용체(androgenic receptor) 단백질의 발현을 억제할 뿐만 아니라, 혈청 및 전립선 내 테스토스테론 및 DHT의 함량을 현저하게 감소시킬 수 있다는 것을 확인함으로써, 본 발명을 완성하였다.The present invention provides a composition for preventing, ameliorating or treating a prostate disease, which comprises an extract of a large capsule as an active ingredient. The present invention provides a composition for preventing, improving or treating a prostate disease, (BPH) to reduce the weight of the enlarged prostate, to inhibit the expression of androgenic receptor protein, and to significantly reduce the content of testosterone and DHT in serum and prostate , Thereby completing the present invention.
상기 목적을 달성하기 위하여, 본 발명은 큰피막이(Hydrocotyle ramiflora) 추출물을 유효성분으로 포함하는 전립선 질환의 예방 또는 치료용 약학 조성물을 제공한다.In order to achieve the above object, the present invention provides greater film (Hydrocotyle The present invention also provides a pharmaceutical composition for prevention or treatment of prostate disease, which comprises an extract of Ramiflora as an active ingredient.
또한, 본 발명은 상기 약학 조성물을 유효성분으로 포함하는 전립선 비대증에 의한 배뇨장애 개선제를 제공한다.The present invention also provides an agent for improving dysuria caused by hypertrophy of the prostate, which comprises the above pharmaceutical composition as an active ingredient.
또한, 본 발명은 큰피막이(Hydrocotyle ramiflora) 추출물을 유효성분으로 포함하는 전립선 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다.In addition, the present invention is based on the finding that a large coating ( Hydrocotyle The present invention also provides a health functional food composition for preventing or ameliorating a prostate disease.
본 발명은 큰피막이 추출물을 유효성분으로 포함하는 전립선 질환의 예방, 개선 또는 치료용 조성물에 관한 것으로, 본 발명의 유효성분인 큰피막이 추출물은 전립선 비대(BPH)에 의해 비후화된 전립선의 무게를 감소시키며, 안드로겐 수용체(androgenic receptor) 단백질의 발현을 억제할 뿐만 아니라, 혈청 및 전립선 내 테스토스테론 및 DHT의 함량을 현저하게 감소시키므로, 전립선 비대증, 전립선염 또는 전립선암의 전립선 질환에 효과적으로 적용할 수 있다. The present invention relates to a composition for preventing, ameliorating or treating a prostate disease, which comprises an extract as an active ingredient, wherein the extract of the large capsule, which is an active ingredient of the present invention, contains the weight of the enlarged prostate by the prostate hypertrophy (BPH) And inhibit the expression of androgenic receptor proteins as well as significantly reduce the levels of testosterone and DHT in the serum and prostate so that they can be effectively applied to prostate diseases of prostate hyperplasia, prostatitis or prostate cancer .
도 1은 본 발명의 큰피막이 추출물의 처리에 따른 세포생존률(%)을 확인한 결과이다. NC는 아무것도 처리하지 않은 음성대조군이다.
도 2는 큰피막이 추출물의 안드로겐 수용체(androgenic receptor; AR) 단백질의 발현억제 효과를 확인한 웨스턴 블랏 결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.
도 3은 도 2의 웨스턴 블랏 결과를 정량화한 결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.
도 4는 본 발명의 큰피막이 추출물의 투여에 따른 전립선의 무게변화를 확인한 결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.
도 5는 본 발명의 큰피막이 추출물의 투여에 따른 혈청 내 DHT 함량 변화를 확인한 결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.
도 6은 본 발명의 큰피막이 추출물의 투여에 따른 혈청 내 테스토스테론의 함량 변화를 확인한 결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.
도 7은 본 발명의 큰피막이 추출물의 투여에 따른 전립선 조직의 H&E 염색결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.
도 8은 본 발명의 큰피막이 추출물의 투여에 따른 간에서의 ALT 및 AST 함량 변화를 확인한 결과이다. NC는 음성대조군; BPH는 전립선 비대증 유발군; BPH+HR는 전립선 비대증 유발 및 큰피막이 추출물 처리군; BPH+F는 전립선 비대증 유발 및 양성대조군인 피나스테리드(Finasteride) 처리군이다.FIG. 1 shows the result of confirming cell survival rate (%) according to the treatment of the extract of the present invention. NC is a negative control with no treatment.
FIG. 2 shows Western blotting results confirming the effect of inhibiting the expression of androgenic receptor (AR) protein of the extracts of large capsules. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 3 is a result of quantifying the Western blot result of FIG. 2. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 4 is a result of checking the weight change of the prostate according to the administration of the extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 5 is a graph showing the change in DHT content in serum according to administration of the extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 6 is a graph showing a change in the content of testosterone in serum according to administration of the extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 7 shows H & E staining results of prostate tissue according to the administration of the extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 8 is a graph showing changes in ALT and AST contents in liver according to the administration of the extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + HR induces hyperplasia of the prostate and treatment of the large capsule with extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
본 발명은 큰피막이(Hydrocotyle ramiflora) 추출물을 유효성분으로 포함하는 전립선 질환의 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention is based on the finding that a large coating ( Hydrocotyle The present invention relates to a pharmaceutical composition for prevention or treatment of prostate diseases.
상기 전립선 질환은 전립선 비대증, 전립선암 또는 전립선염인 것이 바람직하지만 이에 한정하지 않으며, 더 바람직하게는 전립선 비대증이고, 전립선 비대증에 의한 탈모 또는 배뇨장애를 포함할 수 있다. The prostate disease is preferably benign prostatic hyperplasia, prostate cancer or prostatitis, but it is not limited thereto, more preferably it is benign prostatic hyperplasia, and may include hair loss or urination disorder due to enlarged prostate.
상기 큰피막이 추출물은 하기의 단계를 포함하는 방법에 의해 제조할 수 있으나, 이에 한정하지 않는다:The large capsule extract can be prepared by a method including, but not limited to, the following steps:
(1) 큰피막이에 추출용매를 가하여 추출하는 단계;(1) extracting a large coating film with an extraction solvent;
(2) 단계 (1)의 추출물을 여과하는 단계; 및 (2) filtering the extract of step (1); And
(3) 단계 (2)의 여과한 추출물을 감압 농축하고 건조하여 추출물을 제조하는 단계. (3) The step of extracting the filtered extract of step (2) by concentration under reduced pressure and drying.
상기 단계 (1)에서 추출용매는 물, C1~C4의 저급 알코올 또는 이들의 혼합물 중에서 선택하는 것이 바람직하며, 더 바람직하게는 C1~C4의 저급 알코올이고, 더욱더 바람직하게는 99%(v/v) 메탄올이지만 이에 한정하지 않는다.In the step (1), the extraction solvent is preferably selected from water, a C 1 -C 4 lower alcohol or a mixture thereof, more preferably a C 1 -C 4 lower alcohol, even more preferably 99% (v / v) methanol, but is not limited thereto.
상기 제조방법에 있어서, 추출방법은 여과법, 열수 추출, 침지 추출, 환류 냉각 추출 및 초음파 추출 등의 당 업계에 공지된 모든 통상적인 방법을 이용할 수 있다. 상기 추출용매는 건조된 큰피막이 중량의 1~20배 첨가하여 추출하는 것이 바람직하며, 더 바람직하게는 5~15배 첨가하는 것이다. 추출온도는 4 내지 50℃인 것이 바람직하나 이에 한정하지 않는다. 또한, 추출시간은 0.5~10시간인 것이 바람직하며, 0.5~5시간이 더욱 바람직하나 이에 한정하지 않는다. 상기 방법에 있어서, 단계 (3)의 감압농축은 진공 감압 농축기 또는 진공회전증발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한, 건조는 감압건조, 진공건조, 비등건조, 분무 건조 또는 동결 건조하는 것이 바람직하나 이에 한정하지 않는다.In the above production method, any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used. The extraction solvent is preferably extracted by adding 1 to 20 times the weight of the dried large film, more preferably 5 to 15 times the weight of the dried solvent. The extraction temperature is preferably 4 to 50 DEG C, but is not limited thereto. The extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but not always limited thereto. In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in the step (3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.
상기 큰피막이 추출물은 안드로겐 수용체(androgenic receptor) 단백질의 발현을 억제할 수 있으며, 혈청 내 DHT(dihydrotestosterone) 또는 테스토스테론(testosterone)의 함량을 감소시킬 수 있다. The large capsule extract can inhibit the expression of androgenic receptor protein and decrease the content of DHT (dihydrotestosterone) or testosterone in the serum.
본 발명의 약학 조성물은 상기 큰피막이 추출물 이외에 추가로 담체, 부형제 또는 희석제를 더 포함할 수 있다. The pharmaceutical composition of the present invention may further contain a carrier, an excipient or a diluent in addition to the above-mentioned large coating.
본 발명의 약학 조성물은 경구 또는 비경구로 투여될 수 있으며, 비경구 투여 시 피부 외용 또는 복강 내, 직장, 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내 주사 방식을 선택하는 것이 바람직하다.The pharmaceutical composition of the present invention may be administered orally or parenterally, and it is preferable to use an intravenous or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebral injection method during parenteral administration.
본 발명의 약학 조성물은 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 경구투여를 위한 고형 제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용된다. 경구 투여를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성 용제, 현탁제, 유제, 동결건조제제, 좌제가 포함된다. 비수성 용제 및 현탁 용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈tween) 61, 카카오지, 라우린지, 글리세로 젤라틴 등이 사용될 수 있다.The pharmaceutical composition of the present invention can be prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppositories include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
본 발명에 따른 조성물은 약제학적으로 유효한 양으로 투여한다. 본 발명에 있어서, "약제학적으로 유효한 양"은 의학적 치료에 적용 가능한 합리적인 수혜/위험 비율로 질환을 치료하기에 충분한 양을 의미하며, 유효용량 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여 시간, 투여 경로 및 배출 비율, 치료기간, 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 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" 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 type of disease, severity, , 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 can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in 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 side effects, which can be easily determined by those skilled in the art.
본 발명의 조성물의 투여량은 환자의 체중, 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설률 및 질환의 중증도에 따라 그 범위가 다양하게 사용할 수 있다.The dosage of the composition of the present invention may be varied depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease.
또한, 본 발명은 상기 약학 조성물을 유효성분으로 포함하는 전립선 비대증에 의한 배뇨장애 개선제에 관한 것이다.The present invention also relates to an agent for improving dysuria caused by hypertrophy of the prostate, which comprises the above pharmaceutical composition as an active ingredient.
또한, 본 발명은 큰피막이(Hydrocotyle ramiflora) 추출물을 유효성분으로 포함하는 전립선 질환의 예방 또는 개선용 건강기능식품 조성물에 관한 것이다.In addition, the present invention is based on the finding that a large coating ( Hydrocotyle The present invention relates to a health functional food composition for preventing or ameliorating a prostate disease.
상기 큰피막이 추출물을 유효성분으로 포함하는 전립선 질환의 예방 또는 개선용 건강기능식품 조성물은 음료, 환, 정제(tablet), 캡슐제(capsule), 산제 중에서 선택된 어느 하나로 제조하거나, 식품의 성분으로 첨가하여 제조될 수 있으며, 통상적인 방법에 따라 적절하게 제조될 수 있다. The health functional food composition for preventing or ameliorating a prostate disease, wherein the large capsule contains the extract as an active ingredient, may be prepared from any one selected from a beverage, a ring, a tablet, a capsule and a powder, And can be suitably produced according to a conventional method.
본 발명의 큰피막이 추출물을 첨가할 수 있는 식품의 일례로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알코올 음료 및 비타민 복합제 중에서 선택된 어느 하나의 형태일 수 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다.Examples of foods to which the extract of the present invention can be added include meat products, sausages, breads, chocolate, candies, snacks, confectionery, pizza, ramen noodles, other noodles, dairy products including ice- , A tea, a drink, an alcoholic beverage, and a vitamin complex, and includes all the health functional foods in the conventional sense.
상기 건강기능식품은 여러 가지 영양제, 비타민, 광물(전해질), 합성 및 천연 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알킨산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 천연 과일 주스 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. The health functional foods include various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, alkynic acid and its salts, , pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices and vegetable drinks. These components may be used independently or in combination.
본 발명의 건강기능식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다.
The health functional food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient. The natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.
이하, 실시예를 이용하여 본 발명을 더욱 상세하게 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로 본 발명의 범위가 이들에 의해 제한되지 않는다는 것은 당해 기술분야에서 통상의 지식을 가진 자에게 있어 자명한 것이다.
Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.
실시예Example 1. One. 큰피막이Big coat (( 전초outpost , 037-085) 추출물의 제조, 037-085) Preparation of extract
한국식물추출물은행(Korea Plant Extract Bank)으로부터 99.9%(v/v)의 메탄올(HPLC grade)을 추출용매로 사용하여 14.5%의 추출 수율로 획득한 22.26mg의 큰피막이 추출물을 분양받아, 222.6㎕의 DMSO에 녹여 100mg/㎖의 농도로 제조하여 세포 실험(in vitro)에 사용하였다.The extract of 22.26 mg obtained from Korea Plant Extract Bank at an extraction yield of 99.5% (v / v) methanol (HPLC grade) as an extraction solvent was obtained at an extraction yield of 14.5% Of DMSO at a concentration of 100 mg / ml and used for in vitro experiments.
또한, 한국식물추출물은행(Korea Plant Extract Bank)으로부터 99.9%(v/v)의 메탄올(HPLC grade)을 추출용매로 사용하여 20.23%의 추출 수율로 대량 추출한 7.24g의 큰피막이 추출물을 분양받아, 동물 실험(in vivo)에 사용하였다.
In addition, a 7.24 g large-sized membrane extract was obtained from Korean Plant Extract Bank at a extraction yield of 20.23% using 99.9% (v / v) methanol grade (HPLC grade) It was used for animal experiments ( in vivo ).
실시예Example 2. 큰피막이 추출물의 처리에 따른 세포 2. Cells treated with extracts of large capsules 생존률(cell viability)의Of cell viability 확인 Confirm
큰피막이 추출물이 LN cap 세포에 미치는 영향을 확인하기 위하여, 96웰 플레이트에 1×104 개의 LN cap 세포를 도말하여 24시간 동안 37℃에서 배양한 후, 상기 배양한 세포에 농도별(0, 50, 100, 200㎍/㎖)로 큰피막이 추출물을 처리하여 72시간 동안 더 배양하였다. In order to examine the effect of the extract on the LN cap cells, 1 × 10 4 LN cap cells were plated on a 96-well plate and cultured at 37 ° C. for 24 hours. Then, 50, 100, 200 占 퐂 / ml), and the cells were further cultured for 72 hours.
이 후, 웰 당 10㎕의 EZ Cytox Assay Kit(DOGEN, Korea)를 넣고, 2시간 가량 더 반응시킨 후 ELISA(enzyme-linked immunosorbent assay) 리더로 450nm에서 흡광도를 측정하였다.After that, 10 당 of EZ Cytox Assay Kit (DOGEN, Korea) was added to each well, and reacted for about 2 hours. Absorbance was measured at 450 nm with an enzyme-linked immunosorbent assay (ELISA) reader.
그 결과, 도 1에 개시한 바와 같이 큰피막이 추출물을 처리하여도 세포 생존률(cell viability)에는 영향을 주지 않아 세포독성이 거의 없다는 것을 확인하였다.
As a result, as shown in Fig. 1, it was confirmed that even if the large coating film treated with the extract had no effect on cell viability, there was almost no cytotoxicity.
실시예Example 3. 3. 큰피막이Big coat 추출물의 안드로겐 수용체(androgenic receptor; AR) 단백질의 발현억제 효과 확인 The inhibitory effect of extracts on the expression of androgenic receptor (AR) protein
5×105 개의 세포를 6웰 플레이트에 도말하고 24시간 동안 배양한 후, 대조군(control)을 제외한 모든 웰에 100nM의 테스토스테론(testosterone)을 처리하고, 큰피막이 추출물을 농도별(50, 100 및 200㎍/㎖)로 처리한 후, 72시간 동안 37℃, 5% CO2 환경에서 배양하였다. 양성대조군으로 10μM의 피나스테리드(Finasteride)를 사용하였다. 5 × 10 5 cells were plated on a 6-well plate and cultured for 24 hours. All wells except the control were treated with 100 nM testosterone, and the large-capsular extract was treated with concentrations of 50, 100, 200 / / ml), and then cultured in a 5% CO 2 environment at 37 캜 for 72 hours. As a positive control, 10 μM Finasteride was used.
반응이 끝난 후 PBS로 2번 세척하고 단백질 추출용매인 RIPA 완충용액(20mM Tris-HCl, 150mM NaCl, 1mM Na2EDTA, 1mM EGTA, 1% NP-40, 1% 소듐 디옥시콜레이트(sodium deoxycholate), 2.5mM 피로인산나트륨(sodium pyrophosphate), 1mM b-글리세로포스페이트(b-glycerophosphate), 1mM Na3VO4, 1㎍/㎖ 루펩틴(leupeptin; Cell signaling, USA)를 처리하고, 초음파 처리한 후, 얼음에 30분 동안 담아두고 12,000rpm, 20분, 4℃에서 원심분리하였다. 이 후, 상층액만 취하여 단백질 분석키트(Bio-Rad protein assay kit, Bio-Rad , USA)로 단백질을 정량하고 10㎍의 안드로겐 수용체 단백질을 사용하여 웨스턴 블랏을 실시하였다. After the reaction was completed, the cells were washed twice with PBS, and a protein extraction solvent (20 mM Tris-HCl, 150 mM NaCl, 1 mM Na 2 EDTA, 1 mM EGTA, 1% NP- 40, 1% sodium deoxycholate, , 2.5 mM sodium pyrophosphate, 1 mM b-glycerophosphate, 1 mM Na 3 VO 4 , 1 μg / ml leupeptin (Cell signaling, USA) After centrifugation at 12,000 rpm for 20 minutes at 4 ° C, the supernatant was collected and the protein was quantified with a protein analysis kit (Bio-Rad protein assay kit, Bio-Rad, USA) And Western blotting was carried out using 10 [micro] g of the androgen receptor protein.
그 결과, 웨스턴 블랏을 통해 단백질 발현 정도를 확인하고(도 2), 발현량을 정량하여 그래프로 나타내었다. 피막이 추출물을 100㎍/㎖ 농도로 처리했을 때부터 AR의 발현이 감소하였으며, 특히 200㎍/㎖의 농도에서는 양성대조군인 피나스테리드(Finasteride)를 처리한 것보다 더 많이 감소한 것을 확인하였다(도 3).
As a result, the degree of protein expression was confirmed by Western blotting (FIG. 2), and the amount of expression was quantified and shown in a graph. The expression of AR was decreased from the treatment of the extract with 100 μg / ml of the extract, especially at a concentration of 200 μg / ml (Finasteride) (FIG. 3) .
실시예Example 4. 동물 모델의 제작 및 통계 분석 4. Production and statistical analysis of animal models
10주령의 수컷 Wister 랫트(중앙실험동물)를 1주일 동안 순화시킨 후, 전립선 비대 유발군(BPH)은 3mg/kg의 프로피온산테스토스테론(testosterone propionate; TP)를 피하에 4주 동안 주입하여 모델을 확립하였다. 큰피막이 추출물 투여군은 4주 동안 TP 주입 한 시간 전에 100mg/kg의 큰피막이 추출물을 경구 투여하였으며, 양성대조군은 전립선 비대증 치료제로 사용되고 있는 5α-환원 효소 억제제인 피나스테리드(Finasteride)를 10 mg/kg의 양으로 투여하였다.After 10-week-old male Wister rats (central laboratory animals) were purified for 1 week, the prostate hypertrophy group (BPH) was injected with 3 mg / kg of testosterone propionate (TP) Respectively. The extracts were administered orally at a dose of 100 mg / kg of a large capsule of the extract before the injection of TP for 4 weeks. The positive control group was treated with 10 mg / kg of finasteride, a 5α-reductase inhibitor used as a treatment for benign prostatic hyperplasia Lt; / RTI >
통계학적 분석은 ANOVA로 수행하였으며, #는 정상군(NC)과 비교 시 통계적으로 유의미한 차이가 있는 경우로, P<0.05임을 의미하며; *는 전립선 비대증 유발군(BPH)과 비교 시 통계적으로 유의미한 차이가 있는 경우이고, P<0.05임을 의미한다.
Statistical analysis was performed with ANOVA, with # being statistically significant when compared to normal ( P <0.05); * Indicates a statistically significant difference compared to the BPH group, and P <0.05.
실시예Example 5. 전립선 무게 측정 5. Prostate Weighing
흰쥐에 프로피온산테스토스테론(testosterone propionate; TP)를 피하로 4주동안 주사하여 전립선 비대증을 유발하고 큰피막이 추출물과 전립선 비대증 치료제로 사용 중인 피나스테리드(Finasteride)를 4주 동안 함께 경구 투여하였다. 약물 투여 종료 후 희생시킨 쥐의 전립선을 채취하여 무게를 비교하였다.The rats were injected subcutaneously with the testosterone propionate (TP) for 4 weeks to induce hypertrophy of the prostate gland. The finasteride, which was used as a treatment for the large capsule and the prostate hyperplasia, was orally administered for 4 weeks. After the administration of the drug, sacrificed rat prostate glands were sampled and weighed.
그 결과, 도 4에 개시한 바와 같이, 각 실험군(정상군: NC, 전립선 비대증 유발군: BPH, 전립선 비대증 유발군+큰피막이 추출물 투여군: BPH+HR, 전립선 비대증 유발군+Finasteride 투여군: BPH+F)으로부터 측정한 전립선의 무게는 TP로 전립선 비대증을 유발한 군은 정상군에 비해 그 무게가 증가하였고, 전립선 비대증 유발군에 비해 큰피막이 추출물을 투여한 군에서 전립선의 무게가 유의적으로 감소하였다는 것을 확인하였다.
As a result, as shown in Fig. 4, BPH + HR, prostatic hyperplasia + Finasteride-treated group: BPH + HRP, F), the weight of the prostate gland in the group that caused hypertrophy of the prostate gland increased compared to that of the normal group, and the weight of the prostate gland was significantly decreased Respectively.
실시예Example 6. 전립선의 6. Prostate DHTDHT 변화 change
전립선 비대증 동물 모델에서 혈청 내 DHT(dihydrotestosterone)의 함량을 확인하기 위해 실험 종료 후, 분리한 혈액을 4,000rpm에서 20분 동안 원심분리한 후 상등액을 취해 혈청에서 ELISA 키트(ALPCO,Salem)를 이용하여 DHT(dihydrotestosterone)의 함량을 측정하였다. After the end of the experiment, the separated blood was centrifuged at 4,000 rpm for 20 minutes to check the content of DHT (dihydrotestosterone) in an enlarged prostate animal model. The supernatant was taken from the supernatant and the serum was analyzed by ELISA kit (ALPCO, Salem) The content of DHT (dihydrotestosterone) was measured.
그 결과, 도 5에 개시한 바와 같이, 큰피막이 추출물을 투여한 군에서 혈청 내 DHT(dihydrotestosterone)의 함량이 통계적으로 유의미하게 감소한 것을 확인하였다.
As a result, as shown in FIG. 5, it was confirmed that the content of DHT (dihydrotestosterone) in serum was statistically significantly decreased in the group to which the extract was administered.
실시예Example 7. 혈청 내 7. Serum 테스토스테론(testosterone)의Testosterone 함량 변화 분석 Analysis of content change
전립선 비대증 동물 모델에서 테스토스테론(testosterone)의 함량 변화를 확인하기 위해 실험 종료 후 분리한 혈액을 4,000rpm에서 20분간 원심분리한 후 상등액을 취해 혈청에서 ELISA kit (Cayman, USA)을 이용하여 테스토스테론(testosterone)의 함량을 측정하였다. After the end of the experiment, the separated blood was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was taken. The serum was analyzed by ELISA kit (Cayman, USA) for the testosterone (testosterone ) Were measured.
그 결과, 도 6에 개시한 바와 같이 큰피막이 추출물을 투여한 군에서 혈청 내 테스토스테론(testosterone)의 함량이 통계적으로 유의미하게 감소하였다.
As a result, as shown in FIG. 6, the content of testosterone in serum was statistically significantly decreased in the group to which the extract was administered.
실시예Example 8. 전립선의 조직병리학적 변화 8. Histopathological changes of the prostate gland
실험 종료 후 적출한 전립선을 10%의 중성 완충 포르말린으로 24시간 동안 고정한 후 파라핀으로 포매(paraffin embedding)하였다. 포매한 조직은 4μm의 두께로 박절하여 절편을 제작한 후, H&E[헤마톡실린(hematoxylin, Sigma-Aldrich, USA)과 에오신 와이 (eosin Y, Sigma-Aldrich)]로 염색하고, 봉입액으로 봉입하여 광학현미경으로 검경하였다. After the end of the experiment, the extracted prostate gland was fixed with 10% neutral buffered formalin for 24 hours and paraffin embedding was performed. The embryoid tissue was cut to a thickness of 4 μm and cut into sections and stained with H & E (hematoxylin, Sigma-Aldrich, USA) and eosin Y (Sigma-Aldrich) And examined with an optical microscope.
그 결과, 도 7에 개시한 바와 같이, 전립선 비대증 유발군에서는 상피세포의 높이와 수가 과증식되었으나, 큰피막이 추출물을 투여한 군에서는 이러한 상피세포의 과증식이 억제되었다는 것을 확인하였다.
As a result, as shown in FIG. 7, it was confirmed that the height and number of epithelial cells were increased in the prostatic hyperplasia-induced group, but the proliferation of these epithelial cells was inhibited in the group treated with the large-capsular extract.
실시예Example 9. 혈중 ALT 및 9. Serum ALT and ASTAST 측정 Measure
전립선 비대증 동물 모델에서 큰피막이 추출물의 독성 여부를 확인하기 위해 실험 종료 후 분리한 혈액을 4,000rpm에서 20분 동안 원심분리한 후, 상등액을 취해 혈청에서 ALT와 AST의 함량 변화를 확인하였다. After the end of the experiment, the separated blood was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was taken to determine the contents of ALT and AST in the serum.
그 결과, 도 8에 개시한 바와 같이, 일반적인 간 독성(손상) 지표로 사용되는 ALT 및 AST의 함량이 정상군과 비교하여 전립선 비대증 유발군 및 큰피막이 추출물 투여군 모두에서 통계적으로 유의미한 변화는 관찰되지 않았다.As a result, as shown in FIG. 8, there was no statistically significant change in the contents of ALT and AST used as general liver toxicity (damage) indexes in both the patients with benign prostatic hyperplasia and those with large capsules I did.
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| KR1020170061456A KR101896049B1 (en) | 2017-05-18 | 2017-05-18 | Composition for preventing, improving or treating prostate disease comprising extract of Hydrocotyle ramiflora as effective component |
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| KR (1) | KR101896049B1 (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20160016059A (en) * | 2014-08-01 | 2016-02-15 | 전창진 | Hair growth natural composition for coating of head skin |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20160016059A (en) * | 2014-08-01 | 2016-02-15 | 전창진 | Hair growth natural composition for coating of head skin |
Non-Patent Citations (2)
| Title |
|---|
| Journal of Life Science. 2016. Vol.26, Np.9, pp.1022-1.26. * |
| Planta Medica.1998. Vol.64, pp.477-479 * |
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