KR102266418B1 - Composition for preventing and treating prostate disease comprising extract of bulbils of yam as effective component - Google Patents
Composition for preventing and treating prostate disease comprising extract of bulbils of yam as effective component Download PDFInfo
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- KR102266418B1 KR102266418B1 KR1020190092373A KR20190092373A KR102266418B1 KR 102266418 B1 KR102266418 B1 KR 102266418B1 KR 1020190092373 A KR1020190092373 A KR 1020190092373A KR 20190092373 A KR20190092373 A KR 20190092373A KR 102266418 B1 KR102266418 B1 KR 102266418B1
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- young woman
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- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
본 발명은 영여자 추출물을 유효성분으로 함유하는 전립선 질환 예방, 치료용 조성물에 관한 것으로, 더욱 상세하게는 본 발명의 유효성분인 영여자 추출물은 안드로겐 수용체 (androgenic receptor) 전립선 특이항원 (prostate specific antigen) 또는 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현을 억제 기전을 통해, 전립선 질환 중 전립선비대증 또는 전립선암을 효과적으로 예방 및 치료 할 수 있음을 규명하였다. 따라서 본 발명에 따른 영여자 추출물은 전립선 질환 중 전립선 비대증 또는 전립선 암을 예방, 치료 및 개선 할 수 있는 약학적 조성물 또는 건강기능식품 조성물로 유용하게 사용 할 수 있다. The present invention relates to a composition for the prevention and treatment of prostate disease containing the extract of Young woman as an active ingredient, and more particularly, the extract of Young woman as an active ingredient of the present invention is an androgen receptor and prostate specific antigen. ) or 5 alpha-reductase (5-alpha reductase, 5-αR ) through a mechanism of suppressing the expression of prostatic hyperplasia or prostate cancer of the prostate disease was found to be effectively prevented and treated. Therefore, the extract according to the present invention can be usefully used as a pharmaceutical composition or a health functional food composition capable of preventing, treating, and improving prostate enlargement or prostate cancer among prostate diseases.
Description
발명은 영여자 추출물을 유효성분으로 함유하는 전립선 질환 예방, 치료용 조성물에 관한 것으로, 상세하게는 안드로겐 수용체 (androgenic receptor) 전립선 특이항원 (prostate specific antigen) 또는 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현을 억제함으로써 전립선 질환을 치료 및 예방하는 효과를 가진 천연물을 유효성분으로 함유하는 것을 특징으로 하는 조성물에 관한 것이다. The present invention relates to a composition for the prevention and treatment of prostate disease, which contains an extract of young woman as an active ingredient, and more particularly, an androgen receptor, prostate specific antigen or 5-alpha-reductase (5-alpha reductase). , 5-αR ) by inhibiting the expression of a natural product having the effect of treating and preventing prostate disease relates to a composition comprising as an active ingredient.
의료기술의 발달과 건강유지에 대한 관심이 늘어나면서 평균수명이 점점 증가하고 있고 다양한 사회적 이유로 노년층의 인구비율이 급속히 늘어나고 있다. 사회가 고령화 되면서 노인건강에 대한 문제가 대두되고 있다. 선진국의 경우 85세 이상 노인의 40% 이상이 질병에 시달리고 있음이 보고되면서, 단순히 살아있음을 나타내는 평균수명보다 실제로 활동을 하며 건강한 삶을 영위하는 기간을 나타내는 건강수명이 더 중요한 지표로 사용되고 있다. 이에 따라 평소 건강 지향, 웰빙(well-being) 추구 등에 대한 관심이 기능성 건강기능식품의 구매로 이어지면서 새로운 시장이 형성되고 있다. 전립선비대증은 남성의 삶의 질을 저하시키는 대표적인 질환 중 하나다. 40대부터 서서히 발생해 70대 남성의 70%가 겪고 있으며, 그 수가 매년 증가하고 있다. 대표적인 남성 질환인 전립선 비대증(BPH: benign prostatic hyperplasia)은 전립선이 비정상적으로 커지는 질환으로 50세 이상 남성의 50%, 60세 남성의 60%, 85세 남성의 90%에서 발병하며 비대해진 전립선에의해 요도 저항이 증가하면서, 빈뇨, 잔뇨감, 야간뇨, 요단절, 요절박, 약뇨, 요주저 등의 증상을 동반하는 배뇨장애 증상을 의미한다. 전립선비대증의 정확한 발병기전은 아직 밝혀지지 않았으나 혈액 내 존재하는 테스토스테론이 전립선에서 5-알파환원효소(5α-reductase)에 의해 디하이드로테스토스테론(dihydrotestosterone, DHT) 으로 전환되고 DHT에 의해서 안드로겐 수용체(androgen receptor, AR)이 자극되어 전립선비대를 유발한다는 가설이 지배적이다. 전립선비대증의 경우 치료 가이드라인에 따라 알파차단제, 5ARI, 항콜린제, PDE-5 억제제인 타다라필 등을 병용해 처방하고 있다. 기본적으로 전립선비대증으로 인한 배뇨장애 치료에는 알파차단제가 많이 처방되는데, 그 중에서도 기립성 저혈압 등 부작용이 적은 탐스로신 성분 제제가 가장 많이 사용되고 있다. 이렇듯 전립선의 긴장(tone)을 억제하는 약물과 안드로겐(androgen) 호르몬을 감소시켜 전립선의 확장을 차단하는 약물들이 있으나, 교감신경 차단에 의한 저혈압, 혈관 확장성 두통 등의 부작용이 심해 계속적인 사용이 어려움이 있다. 이에 본 발명자는, 천연 물질을 원료로 이용하여 중독이나 부작용의 우려를 해소하며 전립선 비대증 개선의 효능을 갖는 성분을 찾아내어, 남성갱년기에 의하여 유발되는 다양한 질환을 예방하고 치료할 수 있는 조성물을 개발하고자 노력한 결과, 영여자 추출물이 전립선 암 세포주인 LNCaP cell에서, 안드로겐 수용체 (androgenic receptor) 전립선 특이항원 (prostate specific antigen) 또는 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현 억제하는 효과가 있음을 확인함으로써 본 발명을 완성하였다. With the development of medical technology and increasing interest in maintaining health, the average life expectancy is increasing, and the proportion of the elderly population is rapidly increasing for various social reasons. As the society is aging, the problem of the health of the elderly is emerging. In developed countries, it has been reported that more than 40% of the elderly aged 85 and over suffer from diseases, and healthy lifespan, which indicates the period of active and healthy life, is used as a more important indicator than the average lifespan that simply indicates that they are alive. As a result, interest in health-oriented and well-being has led to the purchase of functional health functional foods, creating a new market. BPH is one of the most common diseases that reduce the quality of life of men. It occurs gradually from the age of 40 and affects 70% of men in their 70s, and the number is increasing every year. Benign prostatic hyperplasia (BPH), a typical male disease, is a disease in which the prostate becomes abnormally enlarged. It occurs in 50% of men over the age of 50, 60% of men over the age of 60, and 90% of men over the age of 85. It refers to the symptoms of dysuria accompanied by symptoms such as frequent urination, feeling of residual urination, nocturia, urinary interruption, urgency, weak urination, and hesitation as the urethral resistance increases. Although the exact pathogenesis of BPH is not yet known, testosterone present in the blood is converted to dihydrotestosterone (DHT) by 5-alpha-reductase in the prostate and androgen receptor (androgen receptor) by DHT. , AR) is stimulated to induce an enlarged prostate. In the case of benign prostatic hyperplasia, according to the treatment guidelines, alpha blockers, 5ARIs, anticholinergics, and tadalafil, a PDE-5 inhibitor, are prescribed in combination. Basically, many alpha blockers are prescribed for the treatment of urination disorders due to benign prostatic hyperplasia, and among them, tamsulosin-based formulations with fewer side effects such as orthostatic hypotension are the most used. As such, there are drugs that suppress the tone of the prostate and drugs that block the expansion of the prostate by reducing androgen hormone. However, due to severe side effects such as hypotension and vasodilatory headache due to sympathetic nerve block, continuous use is not recommended. There are difficulties. Accordingly, the present inventors use natural substances as raw materials to solve the concerns of addiction or side effects, find ingredients having the efficacy of improving prostatic hyperplasia, and develop a composition that can prevent and treat various diseases caused by andropause As a result of our efforts, the effect of Young-yeo extract inhibits the expression of androgen receptor, prostate specific antigen, or 5-alpha reductase (5-αR) in LNCaP cells, a prostate cancer cell line. By confirming that there is, the present invention was completed.
본 발명의 목적은 영여자 추출물을 유효성분으로 포함하는 전립선비대증 예방 또는 치료용 약학적 조성물을 제공하는 것이다. It is an object of the present invention to provide a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising the extract of Young woman as an active ingredient.
본 발명의 다른 목적은 영여자 추출물을 유효성분으로 포함하는 전립선비대증 예방 및 개선용 건강기능식품 조성물을 제공하는 것이다. Another object of the present invention is to provide a health functional food composition for preventing and improving prostatic hyperplasia comprising a young woman extract as an active ingredient.
상기 과제를 해결하기 위하여, 본 발명은 영여자 추출물을 유효성분으로 함유하는 전립선질환 예방 또는 치료용 약학적조성물을 제공한다. In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention or treatment of prostate disease containing a young woman extract as an active ingredient.
본 발명의 일실시예에 있어서, 상기 추출물의 추출용매는 물, C1 내지 C6의 알코올 또는 이들의 혼합물일 수 있으며, 또한 상기 알코올은 에탄올, 메탄올 또는 주정 일 수 있다. 또한 상기 전립선 질환은 전립선비대증 또는 전립선암을 특징으로 한다. In one embodiment of the present invention, the extraction solvent of the extract may be water, a C1 to C6 alcohol or a mixture thereof, and the alcohol may be ethanol, methanol or alcohol. In addition, the prostate disease is characterized by an enlarged prostate or prostate cancer.
본 발명의 일 실시예에 있어서, 상기 영여자 추출물은 전립선 조직에서 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현을 감소시킬 수 있으며, 또한 안드로겐 수용체 (androgen receptor, AR) 또는 전립성 특이항원 (prostate specific antigen, PSA)의 발현을 저해 할 수 있다. In one embodiment of the present invention, the extract of the young woman can reduce the expression of 5-alpha reductase (5-αR) in prostate tissue, and also androgen receptor (androgen receptor, AR) or It can inhibit the expression of prostate specific antigen (PSA).
본 발명의 일시예에 있어서, 상기 영여자 추출물은 전립선 질환의 부작용인 성욕감퇴 또는 발기부전이 감소시킬 수 있다. In one embodiment of the present invention, the young woman extract can reduce the side effects of prostate disease, such as decreased libido or erectile dysfunction.
또한 본 발명은 영여자 추출물을 유효성분으로 함유하는 전립선 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다. In addition, the present invention provides a health functional food composition for the prevention or improvement of prostate disease containing a young woman extract as an active ingredient.
본 발명의 일시예에 있어서, 건강기능식품 조성물에 함유된 상기 영여자 추출물은 상기 추출물의 추출용매는 물, C1 내지 C6의 알코올 또는 이들의 혼합물로 추출될 수 있다. In one embodiment of the present invention, the extract of Young woman contained in the health functional food composition may be extracted with water, C1 to C6 alcohol or a mixture thereof as an extraction solvent of the extract.
본 발명의 실시예 있어서, 상기 전립선 질환은 전립선비대증 또는 전립선암일 수 있다. In an embodiment of the present invention, the prostate disease may be an enlarged prostate or prostate cancer.
본 발명의 일시예에 있어서, 건강기능식품 조성물에 함유된 상기 영여자 추출물은 음료, 환, 정제 (tablet), 캡슐제(capsule) 또는 산제 중에서 선택되는 어느 하나의 제형으로 제조 될 수 있다. In one embodiment of the present invention, the extract of Young woman contained in the health functional food composition may be prepared in any one formulation selected from beverages, pills, tablets, capsules, and powders.
상술한 바와 같이, 본 발명의 영여자 추출물은 세포 독성이 없으며, 전립선 암 세포주인 LNCaP cell에서, 안드로겐 수용체 (androgenic receptor), 전립선 특이항원 (prostate specific antigen) 또는 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현을 억제하는 효과를 나타내므로, 전립선비대증, 전립선암 등과 같은 전립선 질환의 예방 및 치료를 위한 약학 조성물 또는 건강기능식품으로써 유용하게 이용될 수 있다.As described above, the extract of the young woman of the present invention has no cytotoxicity, and in LNCaP cells, a prostate cancer cell line, androgen receptor, prostate specific antigen, or 5 alpha-reductase (5-alpha reductase, 5-αR), since it exhibits an effect of inhibiting the expression, it can be usefully used as a pharmaceutical composition or health functional food for the prevention and treatment of prostate diseases such as benign prostatic hyperplasia and prostate cancer.
도 1은 영여자 추출물의 지표성분 정성분석 결과를 나타낸 것이다.
도 2는 전립선암세포주(LNCaP)에 대한 영여자 추출물의 세포 독성을 확인한 결과를 나타낸 것으로서, 6.25, 12.5, 25, 50, 100, 200, 400 μg/ml의 다양한 농도를 처리하여 확인하였을 때, 6.25 내지 200㎍/ml 까지 세포 생존율 (Cell viability) 을 나타낸 것이다.
도 3은 영여자 추출물 농도에 따른 전립선암세포주에 테스토스테론으로 유도된 전립선특이항원(PSA)의 생성 억제 효과를 역전사-실시간 중합효소 연쇄반응(real-time RT-PCR)이용하여, 확인한 결과를 나타낸 것이다.
도 4는 영여자 추출물이 전립선암세포주에서 안드로겐수용체(AR)의 생성 억제 효과를 역전사-실시간 중합효소 연쇄반응(real-time RT-PCR)을 이용하여, 확인한 결과를 나타낸 것이다.
도 5는 영여자 추출물이 전립선암세포주에서 5 알파-환원효소 (5-alpha reductase, 5-αR) 의 단백질 생성 억제 효과를 확인한, 웨스턴 블롯 (western blot) 결과를 나타낸 것이다. 1 shows the results of qualitative analysis of the index component of the extract of Young woman.
Figure 2 shows the results of confirming the cytotoxicity of the young woman extract against the prostate cancer cell line (LNCaP), when it was confirmed by treating various concentrations of 6.25, 12.5, 25, 50, 100, 200, 400 μg / ml, Cell viability is shown from 6.25 to 200 μg/ml.
Figure 3 shows the results of confirming the effect of inhibiting the production of testosterone-induced prostate-specific antigen (PSA) in prostate cancer cell lines according to the concentration of the young woman extract using the reverse transcription-real-time polymerase chain reaction (real-time RT-PCR); will be.
Figure 4 shows the results of confirming the effect of the extract of Youngjae on the production of androgen receptor (AR) inhibition in prostate cancer cell lines using the reverse transcription-real-time polymerase chain reaction (real-time RT-PCR).
Figure 5 shows the results of western blot confirming the effect of the extract of Youngjae on the inhibition of protein production of 5-alpha-reductase (5-αR) in prostate cancer cell lines.
이하 본 발명을 상세히 설명한다. Hereinafter, the present invention will be described in detail.
본 발명은 영여자 추출물을 유효성분으로 함유하는 전립선 질환 예방 및 치료용 약학적 조성물을 제공한다. The present invention provides a pharmaceutical composition for preventing and treating prostate disease containing the extract of Young woman as an active ingredient.
본 발명에서 용어, “영여자 (零餘子, bulbils of yam)”란, 마의 씨앗 또는 씨를 의미하며, 상기 마는 마과에 속하는 다년생 덩굴식물로 주로 뿌리를 식용으로 사용한다. 상기 영여자는 마의 열매, 주아, 마의 식물체 줄기 겨드랑이에 달리는 영양체로 불리기도 하고 씨뿌리 육성을 목적으로 심는 것으로 알려져 있다. 동의보감에서는 상기 영여자에 대하여 ‘보허하고, 허리와 다리를 튼튼하게 하는 효능과 햇볕에 말린 산약의 효능보다 더 강하다’고 기록하고 있으며, 또한 상기 영여자에는 마의 잎이나 뿌리보다 폴리페놀이 12배, 플로보노이드가 3.4배의 함량으로 존재하는 것으로 조사되었다. As used herein, the term “young woman (零餘子, bulbils of yam)” means seeds or seeds of hemp, which is a perennial vine belonging to the family Hemp and mainly uses its roots for food. The young woman is known to be planted for the purpose of cultivating seed roots, and is also called a nutrient that runs on the side of the stem of a hemp fruit, juah, and a hemp plant stem. In Donguibogam, it is recorded that the young woman is 'stronger than the efficacy of strengthening the waist and legs and the efficacy of sun-dried acid medicine' for the young woman. Also, the young woman contains 12 times more polyphenols than the leaves or roots of hemp. , it was investigated that flavonoids were present in an amount of 3.4 times.
본 발명에서 용어, “예방”이란 본 발명에 따른 약학적 조성물의 투여에 의해 전립선 질환을 억제시키거나, 지연시키는 모든 행위를 의미한다. As used herein, the term “prevention” refers to any action that inhibits or delays prostate disease by administration of the pharmaceutical composition according to the present invention.
본 발명에서 용어, “치료”란 본 발명에 따른 약학적 조성물의 투여에 의해 전립선 질환에 의한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다. As used herein, the term “treatment” refers to any action in which symptoms due to prostate disease are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.
본 발명의 약학적 조성물에 의해 예방 또는 치료될 수 있는 전립선 질환에는 비세균성 전립선염, 전립선비대증 또는 전립선암이며, 이에 제한되는 것은 아니며, 또한 상기 약학적 조성물은 상기 전립선 질환의 부작용인 성욕감퇴 또는 발기부전의 발생 억제, 개선 또는 감소되는 것을 특징으로 한다. Prostate diseases that can be prevented or treated by the pharmaceutical composition of the present invention include, but are not limited to, non-bacterial prostatitis, prostatic hyperplasia or prostate cancer, and the pharmaceutical composition is a side effect of the prostate disease, such as decreased libido or erection. It is characterized in that the occurrence of failure is inhibited, ameliorated or reduced.
상기 “전립선 비대증(Benign prostatic hyperplasia: BPH)” 란 여러 가지 이유에 전립선이 비대해지는 질환으로서, 요도나 방광의 경부, 방광 삼각부 등의 점액 분비선이나, 방뇨도선 등이 비대해지는 질환으로, 요오다 방광, 경부, 방광 삼각부 등의 점액 분비선이나 반요도선 등이 비대해지는 하부요로증상 (lover urinary tract symptoms: LUTS)를 포함한다. 전비선 비대증 환자는 소변을 잘 참지 못하고, 가늘고 힘이 없으며, 소변을 보려해도 한참 머뭇거리나 소변을 다 보고 나서도 남아 있는 듯한 잔뇨감 등의 증상이 나타난다. The "Benign prostatic hyperplasia (BPH)" is a disease in which the prostate is enlarged for various reasons, such as the urethra, the neck of the bladder, the bladder triangle, etc. These include lover urinary tract symptoms (LUTS) in which the mucous glands or semiurethral glands of the bladder, neck, and bladder triangle are enlarged. Patients with an enlarged anterior splenomegaly have symptoms such as difficulty in holding urine, thin and weak, hesitating to urinate for a long time, and a feeling of residual urination that seems to remain even after urinating.
본 발명에서 용어, ‘5 알파-환원효소 (5-alpha reductase, 5-αR)’란 전립선에서 테스토스테론(testosterone)을 전립선 조직의 성자엥 관여하는 주된 남성호르몬인 디하이드로테스토스테론 (dihydrotstosterone, DHT)으로 전환시키는 효소로, 전립선 비대증의 치료에 5 알파-환원효소 억제제 이용은 당업자에 자명한 수준에 이르렀다. 하지만 합성 5 알파-환원효소 억제제 이용은 복용 후 효과가 나타날 때까지 오랜 시간이 걸리고, 성욕감퇴, 발기부전 등의 남성 성기능을 감소시키는 부작용뿐만 아니라, 어지러움, 피로감, 그리고 과도한 저혈압과 같은 부작용이 있어, 천연 물질을 원료로 이용한 신약 개발이 필요하다. As used herein, the term '5 alpha-reductase (5-αR)' refers to dihydrotestosterone (DHT), which is the main male hormone involved in the growth of prostate tissue from testosterone in the prostate. As a converting enzyme, the use of 5 alpha-reductase inhibitors in the treatment of benign prostatic hyperplasia has reached a level apparent to those skilled in the art. However, the use of synthetic 5-alpha-reductase inhibitors takes a long time to take effect after taking them, and there are side effects such as dizziness, fatigue, and excessive hypotension, as well as side effects that reduce male sexual function such as decreased libido and erectile dysfunction. However, it is necessary to develop new drugs using natural substances as raw materials.
본 발명의 따른 영여자 추출물은 하기 단계를 포함하는 방법에 의해 제조 될 수 있으나, 이에 한정되지 않는다. Young woman extract according to the present invention may be prepared by a method comprising the following steps, but is not limited thereto.
(1) 영여자 건조물에 추출용매를 가하여 추출하는 단계; (1) extracting by adding an extraction solvent to the dried young woman;
(2) 단계 (1) 의 추출물을 침지 및 감암농축하는 단계; (2) immersing and dark-concentrating the extract of step (1);
및 (3) 단계 (2)의 감압농축물을 건조하는 단계.and (3) drying the vacuum concentrate of step (2).
상기 단계 (1)에서 추출용매는 물, C1 내지 C6의 저급 알코올 또는 이들의 혼합물 중에서 선택하는 것이 바람직하며, 상기 알코올은 에탄올, 메탄올 또는 주정을 사용하는 것이 더욱 바람직하나 이에 한정되지 않는다. 추출용매의 양은 영여자 중량의 2 내지 15 배로 , 바람직하게는 5 내지 10배 첨가하여 추출하는 것이 바람직하나, 이에 한정되지 않는다. 추출방법은 열 추출, 환류 냉각 추출, 초음파 추출 등의 당업계에 공지된 모든 통상적인 방법을 이용할 수 있다. 또한 추출시간은 0.5 내지 10시간인 것이 바람직하며, 이에 한정되지 않는다. 상기 방법에 있어서, 단계 (3)의 감압농축은 진공 농축기 또는 진공회전증 발기를 이용하는 것이 바람직하나 이에 한정하지 않는다. 또한 건조는 감압건조, 진공건조, 비등건조, 분무건조 또는 동결건조하는 것이 바람직하나, 이에 한정하지 않는다. In step (1), the extraction solvent is preferably selected from water, a C1 to C6 lower alcohol or a mixture thereof, and the alcohol is more preferably ethanol, methanol or alcohol, but is not limited thereto. The amount of the extraction solvent is preferably 2 to 15 times the weight of the young woman, preferably 5 to 10 times the weight of the extract, but is not limited thereto. The extraction method may use all conventional methods known in the art, such as thermal extraction, reflux cooling extraction, ultrasonic extraction, and the like. In addition, the extraction time is preferably 0.5 to 10 hours, but is not limited thereto. In the above method, it is preferable to use a vacuum concentrator or vacuum rotary evaporator for concentration under reduced pressure in step (3), but is not limited thereto. In addition, drying under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying is preferable, but is not limited thereto.
상기 영여자 추출물은 안드로겐 수용체 (androgenic receptor) 전립선 특이항원 (prostate specific antigen) 또는 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현 억제 할 수 있다. The young woman extract may inhibit androgen receptor (androgenic receptor) prostate specific antigen (prostate specific antigen) or 5-alpha-reductase (5-alpha reductase, 5-αR) expression.
본 발명의 약학적 조성물은 상기 영여자 추출물 이외에 추가로 담체, 부형제 또는 희석제를 더 포함할 수 있다. The pharmaceutical composition of the present invention may further include a carrier, excipient, or diluent in addition to the extract of Young woman.
본 발명의 약학적 조성물은 경구 또는 비경구로 투여(예를 들어, 도포 또는 정맥 내 , 피하, 복강 내 주사) 될 수 있으나, 경구 투여가 바람직하다. The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, application or intravenous, subcutaneous, intraperitoneal injection), but oral administration is preferred.
비경구 투여를 위한 제제로는 각각 통상의 방법에 다란 산제, 과립제, 정제, 캡슐제, 멸균된 수용액, 액제, 비수성용제, 현택제, 에멀젼, 시럽, 좌제, 에어로졸 등의 외용제 및 멸균 주사제제의 형태로 제형화하여 사용될 수 있으며, 바람직하게는 크림, 젤, 패취, 분무제, 연고제, 경고제, 로션제, 리니멘트제, 파스타제 또는 카타플라스마제의 피부 외용 약학적 조성물을 제조하여 사용 할 수 있으나, 이에 한정되는 것은 아니다. 국소 투여의 조성물은 임상적 처방에 따라 무수형 또는 수성형 일 수 있다. 비수성용제, 현탁제로는 프로필렌글리콜 (prolylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 이텔올레이트와 같은 주사 가능한 에스테르 등이 사용 될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween)61, 카카오지, 라우린지, 글리세로제라틴 등이 사용 될 수 있다. Formulations for parenteral administration include powders, granules, tablets, capsules, sterilized aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, syrups, suppositories, aerosols, etc. It can be formulated and used in the form of a cream, gel, patch, spray, ointment, warning agent, lotion, liniment agent, pasta agent or cataplasma agent for external use by preparing a pharmaceutical composition for external use. However, the present invention is not limited thereto. Compositions for topical administration may be anhydrous or aqueous, depending on the clinical prescription. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as yteloleate may be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, etc. may be used.
경구투여를 위한 고형제제는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형 제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제 예를 들면 전분, 탄산칼슘, 스크로오스 (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. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include one or more compounds and at least one excipient, for example, starch, calcium carbonate, sucrose or lactose. It is prepared by mixing lactose, gelatin, etc. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid formulations for oral administration include suspensions, solutions, emulsions, syrups, etc., and various excipients, such as wetting agents, sweeteners, and fragrance preservatives, in addition to commonly used simple diluents such as water and liquid paraffin, may be included. have.
본 발명에 따른 조성물은 약제학적으로 유요한 양으로 투여한다. 본 발명에 있어서, “약제학적으로 유효한 양”은 의학적 치료에 적용 가능한 합리적인 수혜. 위험 비율로 질환을 치료하기 위한 충분한 양을 의미하며, 유효용량, 수준은 환자의 질환의 종류, 중증도, 약물의 활성, 약물에 대한 민감도, 투여시간, 투여 경로 및 배출 비율, 치료기간 동시 사용되는 약물을 포함한 요서 및 기타 의학 분야에 잘 알려진 요소에 따라 결정 될 수 있다. 본 발명의 조성물은 개별 치료제로 투여하거나, 다른 치료제와 병용하여 투여될 수 있고, 종재의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업제에 의해 용이하게 결정 될 수 있다. The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" is a reasonable benefit applicable to medical treatment. It refers to an amount sufficient to treat a disease in a risk ratio, and the effective dose and level are the type, severity, drug activity, sensitivity to drug, administration time, administration route and excretion rate of the patient's disease, concurrently used during the treatment period. It can be determined according to factors well known in the field of medicine and other factors, including drugs. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with the conventional therapeutic agents, and may be administered singly or multiple times. In consideration of all of the above factors, it is important to administer an amount capable of obtaining the maximum effect with a minimum 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 사용 할 수 있다. The dosage of the composition of the present invention can be used in various ranges depending on the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate and severity of disease.
본 발명은 영여자 추출물을 유효성분으로 함유하는 전립선 질환의 예방 또는 개선용 건강기능식품 조성물을 제공한다. 상기 전립선 질환은 전립선비대증 또는 전립선암을 특징으로 한다. The present invention provides a health functional food composition for the prevention or improvement of prostate disease containing a young woman extract as an active ingredient. The prostate disease is characterized by an enlarged prostate or prostate cancer.
상기 추출물은 음료, 환, 정제 (tablet), 캡슐제(capsule) 또는 산제 중에서 선택되는 어느 하나의 제형으로 제조하거나 식품의 성분으로 첨가하여 제조 될 수 있으며, 통상적인 방법에 따라 적정하게 제조 될 수 있다. The extract may be prepared in any one formulation selected from beverages, pills, tablets, capsules, or powders, or may be prepared by adding it as a component of food, and may be appropriately prepared according to a conventional method. have.
본 발명의 영여자 추출물을 첨가할 수 잇는 식품의 일례로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차 드링크제, 알코롱 음료 및 비타민 복합체 중에서 선택된 어느 하나의 형태일 수 있으며, 통상적인 의미에서 건강기능식품을 모두 포함한다. 상기 건강기능식품은 여러 가지 영양제, 비타민, 광물(전해질), 합성 및 천연 풍미제, 착색제 및 증진제 (치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 중점제, PH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화 등을 함유할 수 있다. 그 밖에 천연 과일 주스 및 야채음료 제조를 위한 과육을 함 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다.Examples of foods to which the Young woman extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages , tea drinks, alcohol drinks, and vitamin complexes may be in any one form, and includes all health functional foods in a conventional sense. The health functional food includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, and protective colloids. , PH adjuster, stabilizer, preservative, glycerin, alcohol, carbonation used in carbonated beverages, and the like. In addition, the pulp for the manufacture of natural fruit juices and vegetable beverages can be added. These components may be used independently or in combination.
본 발명의 건강기능식품 조성물은 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로 함유할 수 있다. 상기 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 텍스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소트비통, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마린, 스테비아 추출과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 가미제 등을 사용할 수 있다. The health functional food composition of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, Sortbiton, and erythritol. As the sweetener, natural sweeteners such as taumarin and stevia extract, and synthetic flavoring agents such as saccharin and aspartame can be used.
이하, 본 발명을 실시예와 제조예를 통하여 상세히 설명한다. 단 하기의 실시예와 제조예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예와 제조예에 의하여 한정되는 것은 아니다. Hereinafter, the present invention will be described in detail through Examples and Preparation Examples. However, the following examples and preparation examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following examples and preparation examples.
<실시예 1> 영여자 추출물 제조 및 추출물의 성분<Example 1> Preparation of young woman extract and ingredients of the extract
본 발명에 따른 영여자 추출물을 하기와 같이 제조 및 준비하였다. 영여자 건조물에 대하여 에탄올 첨가하여 침지 및 감압농축하여 추출물을 준비하였다. A young woman extract according to the present invention was prepared and prepared as follows. An extract was prepared by adding ethanol to the dried young woman, immersing it and concentration under reduced pressure.
추출용매는 물, C1 내지 C6의 저급 알코올 또는 이들의 혼합물 중에서 선택하는 것이 바람직하며, 상기 알코올은 메탄올 또는 에탄올을 사용하는 것이 더욱 바람직하나 이에 한정되지 않는다. 추출용매의 양은 영여자 중량의 2 내지 15 배로 , 바람직하게는 5 내지 10배 첨가하여 추출하는 것이 바람직하나, 이에 한정되지 않는다. 추출방법은 열 추출, 환류 냉각 추출, 초음파 추출 등의 당업계에 공지된 모든 통상적인 방법을 이용할 수 있다. The extraction solvent is preferably selected from water, a lower alcohol of C1 to C6, or a mixture thereof, and the alcohol is more preferably methanol or ethanol, but is not limited thereto. The amount of the extraction solvent is preferably 2 to 15 times the weight of the young woman, preferably 5 to 10 times the weight of the extract, but is not limited thereto. The extraction method may use all conventional methods known in the art, such as thermal extraction, reflux cooling extraction, ultrasonic extraction, and the like.
본 발명의 전립선 비대증 표지인자 억제 활성을 갖는 영여자 추출물에 대한 정성분석을 UPLC-OTof-MS (액체크로마토그래피 고분해능 탄댐 질량분석기, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry) 분석을 통하여 분석하였다. 상기 영여자 추출물에 대한 지표물질의 정성분석은 도 1과 같다. 상기 정성분석결과, Kaempferol 3-o-(6''-acetyl-galactoside) 7-o-rhamnoside, 2-O-protocatechuoyl alphitolic acid, theaflavin, Myricetin 3',4'-dimethyl ether 3-rhamnoside, RheidinC (one mole of rhein <llld one mole of physcion), Tricin-5-glucoside 등 산화방지 폴리페놀 또는 플라보노이드 성분들이 포함한 총 25종 성분이 확인되었다. UPLC-OTof-MS (liquid chromatography high-resolution tandam mass spectrometry, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry) analysis of the qualitative analysis of the extract of the young woman having the inhibitory activity of the prostatic hyperplasia markers of the present invention was analyzed through Qualitative analysis of the indicator material for the Young woman extract is shown in FIG. 1 . As a result of the above qualitative analysis, Kaempferol 3-o-(6''-acetyl-galactoside) 7-o-rhamnoside, 2-O-protocatechuoyl alphitolic acid, theaflavin, Myricetin 3',4'-dimethyl ether 3-rhamnoside, RheidinC ( A total of 25 components including antioxidant polyphenols or flavonoids such as one mole of rhein <llld one mole of physcion) and Tricin-5-glucoside were identified.
<실시예 2> 영여자의 세포 독성 또는 세포 생존율 (Cell viability) 확인 < Example 2> Confirmation of cytotoxicity or cell viability of young women
전립선암세포주(LNCaP cell)를 우태아혈청(fetal bovine serum; FBS)이 10 % 함유된 RMPI-1640 배지에서 배양하였다. 즉, 175 ㎠ 플라스틱 플라스크(SPL life science Co., Ltd. Korea)에 전립선암세포주를 10 % FBS, 1% 항생제(antibiotic)/항진균제(antimycotics)가 함유된 RPMI-1640 배지에서 37℃, 5% CO₂의 조건 하에서 배양하였다. 2~3일마다 한 번씩 2차 배양하여 세포주를 유지하였다. Prostate cancer cell lines (LNCaP cells) were cultured in RMPI-1640 medium containing 10% fetal bovine serum (FBS). That is, in a 175 cm2 plastic flask (SPL life science Co., Ltd. Korea), the prostate cancer cell line was prepared in RPMI-1640 medium containing 10% FBS and 1% antibiotic/antimycotics at 37° C., 5% It was cultured under the condition of CO₂. Cell lines were maintained by secondary culture once every 2-3 days.
세포가 자란 175 ㎠ 플라스틱 플라스크에서 배지액을 제거하고, CMF-PBS (calcium magnesium free-phosphate buffered saline, pH 7.2)로 세척한 후, 0.25 % 트립신/EDTA를 처리하여 세포를 플라스크 바닥으로부터 떼어내고 세포 배양액으로 중화시킨 후 원심분리(1000 rpm, 3 min) 하였다. 남은 세포의 펠렛(pellet)에 배양액을 가한 다음, 멸균 피펫으로 반복 흡입하여 단일 세포 부유액을 제조하였다. 제조한 세포 부유액과 트립판 블루(trypan blue)를 1:1의 비율로 혼합하여 광학 현미경상에서 혈구계산판(hemocytometer)을 이용하여 측정하는 방법으로 세포 계대를 하여, 세포 독성을 확인을 위한 세포 배양 및 계대를 하였다. The medium was removed from the 175 cm2 plastic flask in which the cells were grown, washed with CMF-PBS (calcium magnesium free-phosphate buffered saline, pH 7.2), and treated with 0.25% trypsin/EDTA to remove the cells from the bottom of the flask and remove the cells. After neutralization with the culture medium, centrifugation (1000 rpm, 3 min) was performed. A single cell suspension was prepared by adding the culture solution to the remaining cell pellet and then repeatedly suctioning with a sterile pipette. The prepared cell suspension and trypan blue are mixed in a ratio of 1:1, and the cells are passaged by measuring using a hemocytometer on an optical microscope, and cell culture to check cytotoxicity. and passaged.
본 발명에 따른 영여자의 전립선암세포주(LNCaP cell)에 대한 독성 측정은 EzCytox 키트를 이용하였다. 구체적으로, 전립선암세포주를 96 well plate에 1×104 cells/well이 되도록 분주하였다. 이를 37℃, 5% CO₂ 조건의 배양기에서 배양한 후, 0, 6.25, 12.5, 25, 50, 100, 200, 400 μg/ml의 다양한 농도의 영여자 추출물을 첨가하여 24시간 동안 배양하였다. 24시간 동안 배양한 전립선암세포주를 대상으로 EzCytox키트를 이용하여 1시간 동안 반응시킨 후 흡광도를 측정하였다. 대조군을 기준으로 세포 생존율을 계산함으로써 영여자의 세포 독성을 결정하였다. 그 결과, 도 2에 나타낸 바와 같이, 영여자를 첨가하지 않았을 경우 전립선암세포주는 100 %의 생존률을 나타내었으며, 영여자를 여러 농도로 첨가한 군에서는 100 μg/ml의 농도까지 90% 이상의 생존율을 보였다. 따라서 본 발명의 영여자 추출물은 전립선암 세포주에 대하여 세포독성을 나타내지 않음을 확인하였다(도 2).The EzCytox kit was used to measure the toxicity of the prostate cancer cell line (LNCaP cell) of the young woman according to the present invention. Specifically, the prostate cancer cell line was aliquoted in a 96-well plate to 1×10 4 cells/well. After culturing them in an incubator at 37° C. and 5% CO₂, 0, 6.25, 12.5, 25, 50, 100, 200, 400 μg/ml of Young woman extracts of various concentrations were added and cultured for 24 hours. Prostate cancer cell lines cultured for 24 hours were reacted for 1 hour using the EzCytox kit, and then absorbance was measured. Cytotoxicity of infants was determined by calculating cell viability relative to the control group. As a result, as shown in FIG. 2 , the prostate cancer cell line showed a 100% survival rate when no young woman was added, and a survival rate of 90% or more up to a concentration of 100 μg/ml in the group in which the young woman was added at various concentrations. seemed Therefore, it was confirmed that the extract of the present invention does not show cytotoxicity against prostate cancer cell lines (FIG. 2).
<실시예 3> 테스토스테론(testosterone)으로 유도된 전립선특이항원(prostate specific antigen, PSA)에 대한 영여자 추출물의 농도별 억제 효과 확인<Example 3> Confirmation of inhibitory effect by concentration of young woman extract on testosterone-induced prostate specific antigen (PSA)
테스토스테론(testosterone)으로 유도된 전립선암세포주(LNCaP cell)에서의 전립선특이항원(PSA)의 발현 억제 측정은 역전사-실시간 중합효소 연쇄반응(real-time RT-PCR)를 이용하였다. 구체적으로, 전립선암세포주를 6 well plate에 5×105 cells/well이 되도록 분주하였다. 이를 37℃, 5% CO₂조건의 배양기에서 24시간 배양한 후, 0 (대조군), 200, 100, 50 μg/ml의 다양한 농도의 영여자 추출물을 첨가하여 3시간 동안 배양하였다. testosterone 100 nM 을 대조군을 제외한 각 well에 첨가하고 24시간 동안 배양하였다. 24시간 동안 배양한 후, 상층액을 제거하고 Trizol을 사용하여 total RNA을 추출한 후 PrimeScript RT Master Mix(Takara, Japan)을 사용하여 cDNA를 합성하였다. 합성된 cDNA는 TOPreal qPCR 2X PreMIX(Enzynomics, Korea)를 이용하여 실시간 중합효소 연쇄반응을 진행하였다. 상기 실시간 중합효소 연쇄반응 조건은 Hat start (Pre-denaturation) 95℃ 12 분 (min), Denaturation 95℃ 10 초(sec). Annealing 60℃ 30sec(초) 이고, 사용한 프라이머 (primer) 서열은 하기 표 1과 같다.In the testosterone-induced prostate cancer cell line (LNCaP cell), the expression inhibition of the prostate-specific antigen (PSA) was measured by reverse transcription-real-time polymerase chain reaction (real-time RT-PCR). Specifically, the prostate cancer cell line was aliquoted in a 6-well plate to 5×10 5 cells/well. After culturing for 24 hours in an incubator at 37° C. and 5% CO₂, 0 (control), 200, 100, and 50 μg/ml of Young woman extracts of various concentrations were added and incubated for 3 hours. 100 nM of testosterone was added to each well except the control and cultured for 24 hours. After incubation for 24 hours, the supernatant was removed, total RNA was extracted using Trizol, and cDNA was synthesized using PrimeScript RT Master Mix (Takara, Japan). The synthesized cDNA was subjected to real-time polymerase chain reaction using TOPreal qPCR 2X PreMIX (Enzynomics, Korea). The real-time polymerase chain reaction conditions are Hat start (Pre-denaturation) 95
< 전립선 특이항원 (PSA) 억제 효과 확인을 위한 프라이머 목록>< List of primers for confirming the effect of inhibiting prostate-specific antigen (PSA)>
antigen (PSA)Prostate specific
antigen (PSA)
그 결과, 도 3에 나타낸 바와 같이, 영여자를 처리 하지 않았을 경우 전립선암세포주에서 testosterone에 의해 PSA의 발현이 약 200% 증가하였으며, testosterone 만 처리한 군 대비 영여자 추출물을 50, 100, 200 μg/ml 첨가하였을 때 각각 35, 50, 88 % 의 PSA 발현 억제능을 보임을 확인하였다 (도 3).As a result, as shown in FIG. 3 , when no young woman was treated, the expression of PSA was increased by about 200% by testosterone in prostate cancer cell lines, and 50, 100, 200 μg of young woman extract compared to the testosterone-treated group was used. When /ml was added, it was confirmed that 35, 50, and 88% of PSA expression inhibition ability was shown (FIG. 3).
<실험예 4> 영여자 추출물의 안드로겐 수용체(Androgen receptor, AR) 발현 저해 효과 확인<Experimental Example 4> Confirmation of androgen receptor (AR) expression inhibitory effect of young woman extract
전립선암세포주(LNCaP cell)에서의 안드로겐 수용체(Androgen receptor, AR)의 발현 양 측정은 역전사-실시간 중합효소 연쇄반응(real-time RT-PCR)를 이용하였다. 구체적으로, 전립선암 세포주를 6 well plate에 5×105 cells/well이 되도록 분주하였다. 이를 37℃, 5% CO₂ 조건의 배양기에서 24시간 배양한 후, 0 (대조군), 200, 100, 50 μg/ml의 다양한 농도의 영여자 추출물을 첨가하여 3시간 동안 배양하였다. testosterone 100 nM 을 대조군을 제외한 각 well에 첨가하고 24시간 동안 배양하였다. 배양이 끝난 후, 상층액을 제거하고 Trizol을 사용하여 total RNA을 추출한 후 PrimeScript RT Master Mix(Takara, Japan)을 사용하여 cDNA를 합성하였다. 합성된 cDNA는 TOPreal qPCR 2X PreMIX(Enzynomics, Korea)를 이용하여 실시간 중합효소 연쇄반응을 진행하였다. PCR 조건은 Hat start (Pre-denaturation) 95℃ 12 분 (min), Denaturation 95℃ 10 초(sec). Annealing 60℃ 30sec(초) 이고, 사용한 프라이머 (primer)는 하기의 표 2와 같다. The expression level of androgen receptor (AR) in prostate cancer cell lines (LNCaP cells) was measured using reverse transcription-real-time RT-PCR. Specifically, the prostate cancer cell line was aliquoted in a 6-well plate to 5×10 5 cells/well. After culturing for 24 hours in an incubator at 37°C and 5% CO₂ conditions, 0 (control), 200, 100, and 50 μg/ml of Young woman extract at various concentrations were added and incubated for 3 hours. 100 nM of testosterone was added to each well except the control and cultured for 24 hours. After incubation, the supernatant was removed, total RNA was extracted using Trizol, and cDNA was synthesized using PrimeScript RT Master Mix (Takara, Japan). The synthesized cDNA was subjected to real-time polymerase chain reaction using TOPreal qPCR 2X PreMIX (Enzynomics, Korea). PCR conditions are Hat start (Pre-denaturation) 95
<안드로겐 수용체 발현 저해 효과를 확인하기 위한 프라이머 (primer) 목록><List of primers to confirm the effect of inhibiting androgen receptor expression>
(AR)Androgen receptor
(AR)
그 결과, 도 4에 나타낸 바와 같이, 영여자의 열매추출물을 처리 하지 않았을 경우 전립선암세포주에서 testosterone에 의해 AR의 발현이 약 140% 증가하였으며 영여자 추출물을 50, 100, 200 μg/ml 첨가하였을 때 각각 19, 20, 34% 의 AR 발현 억제능을 보임을 확인하였다 (도 4).As a result, as shown in FIG. 4, when the fruit extract of Young woman was not treated, AR expression was increased by about 140% by testosterone in prostate cancer cell lines, and 50, 100, and 200 μg/ml of Young woman extract were added. When it was confirmed that 19, 20, 34% of AR expression inhibition ability, respectively (FIG. 4).
<실험예 5> 영여자 추출물 처리군에서 5-alpha reductase 단백질 발현 억제 효과 확인<Experimental Example 5> Confirmation of the effect of inhibiting 5-alpha reductase protein expression in the young woman extract treatment group
전립선암세포주(LNCaP cell)에서의 5-alpha reductase(5-αR)의 발현 양 측정은 western blot assay를 이용하였다. 상기 western blot assay은 Phosphate buffered saline (PBS)에 SDS(Sodium dodecyl sulfate)를 녹여 1% SDS buffer를 제작하고, 1% SDS buffer 에 Protease Inhibitor Cocktail(Roche, Mannheim, Germany)와 PhosphoStop(Roche, Mannheim, Germany)을 첨가하여 protein lysis buffer를 제작하였다. 세포에 protein lysis buffer를 처리하여 단백질을 추출하고 sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)에 이동시켜 단백질 크기별로 분리한 뒤 Poly vinylidene fluoride (PVDF) membrane에 부착시킨다. membrane에 부착된 단백질들은 gene specific antibody 들을 이용하여 발현 양을 측정하였다. 더욱 구체적으로, 전립선암세포주를 6 well plate에 5×105 cells/well이 되도록 분주하였다. 이를 37℃, 5% CO₂ 조건의 배양기에서 24시간 배양한 후, 0 (대조군), 200, 100, 50 μg/ml의 다양한 농도의 영여자 추출물을 첨가하여 3시간 동안 배양하였다. testosterone 100 nM 을 대조군을 제외한 각 well에 첨가하고 24시간 동안 배양하였다. 24시간 동안 배양한 후, 상기와 같이 Western blot assay을 실시하여 5-alpha reductase 의 발현을 측정하였다. 그 결과, 도 5와 같이, 영여자 추출물을 첨가함에 따라 5-alpha reductase 의 발현이 현저히 감소됨을 확인하였다 ( 도5).Western blot assay was used to measure the expression level of 5-alpha reductase (5-αR) in prostate cancer cell lines (LNCaP cells). remind For western blot assay, 1% SDS buffer is prepared by dissolving sodium dodecyl sulfate (SDS) in phosphate buffered saline (PBS), and Protease Inhibitor Cocktail (Roche, Mannheim, Germany) and PhosphoStop (Roche, Mannheim, Germany) in 1% SDS buffer. ) was added to prepare a protein lysis buffer. Proteins are extracted by treating the cells with protein lysis buffer, transferred to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), separated by protein size, and attached to a polyvinylidene fluoride (PVDF) membrane. Proteins attached to the membrane were measured for expression using gene specific antibodies. More specifically, 5 × 10 prostate cancer cell lines in a 6 well plate5Cells/well were dispensed. After culturing for 24 hours in an incubator at 37°C and 5% CO₂ conditions, 0 (control), 200, 100, and 50 μg/ml of Young woman extract at various concentrations were added and incubated for 3 hours. 100 nM of testosterone was added to each well except the control and cultured for 24 hours. After incubation for 24 hours, the expression of 5-alpha reductase was measured by performing Western blot assay as described above. As a result, as shown in FIG. 5 , it was confirmed that the expression of 5-alpha reductase was remarkably reduced as the Young woman extract was added ( FIG. 5 ).
하기의 본 발명의 추출물 함유하는 약학적 조성물 및 건강기능식품 조성물에 대한 제조예를 설명하나, 이는 본 발명을 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다. Preparation examples for the pharmaceutical composition and health functional food composition containing the extract of the present invention are described below, but this is not intended to limit the present invention, but only to describe in detail.
<제조예 1> 약학적 제제의 제조<Preparation Example 1> Preparation of pharmaceutical formulations
1. 정제의 제조1. Preparation of tablets
상기 실시예에 수득한 본 발명의 영여자 추출물을 유효성분으로 함유하는 정제는 하기와 같은 방법으로 제조하였다. 락토오스, 전분 및 전젤라티환 옥수수 전분을 영여자 추출물과 혼합한 후, 적합한 용적의 정제수를 첨하하고, 분말로 과립화시켰다. 과립을 건조시킨 후 스테아르산 마그네슘과 혼합하고 압착하여 정제를 제조하였다. 상기 정제의 구성성분은 하기와 같다. The tablet containing the extract of the present invention obtained in the above Example as an active ingredient was prepared as follows. After mixing lactose, starch and whole-gelatih corn starch with the extract of Young woman, an appropriate volume of purified water was added thereto, and granulated into a powder. After drying the granules, they were mixed with magnesium stearate and compressed to prepare tablets. The components of the tablet are as follows.
영여자 추출물 5.0 mgYoung woman extract 5.0 mg
락토오스 BP 150.0 mgLactose BP 150.0 mg
전분 BP 30.0 mgStarch BP 30.0 mg
전젤라틴화 옥수수 전분 BP 15.0 mgPregelatinized Corn Starch BP 15.0 mg
스테아르산 마그네슘 1.0 mg1.0 mg magnesium stearate
2. 캡슐제의 제조2. Preparation of capsules
본 발명의 영여자 추출물을 유효성분으로 함유하는 캡슐제는 하기와 같은 방법으로 제조하였다. 상기 본 발명의 영여자 추출물의 일정량의 부형제 및 스테아르산 마그네슘과 혼합하였다. 얻어진 혼합물을 젤라틴 캡슐 중에 충전하여 캡슐을 제조하였다.A capsule containing the extract of the present invention as an active ingredient was prepared as follows. A certain amount of the excipient and magnesium stearate of the extract of the present invention was mixed. The obtained mixture was filled in gelatin capsules to prepare capsules.
상기 캡슐제의 구성성분은 하기와 같다. The components of the capsule are as follows.
영여자 추출물 5.0 mgYoung woman extract 5.0 mg
전분 1500 100.0 mgStarch 1500 100.0 mg
스테아르산 마그네슘 BP 1.0 mgMagnesium stearate BP 1.0 mg
3. 주사제의 제조3. Preparation of injections
본 발명의 영여자 추출물을 유효성분으로 함유하는 주사제는 하기와 같은 방법으로 제조하였다. An injection containing the extract of the present invention as an active ingredient was prepared as follows.
적당한 용적의 주사용 염화나트륨 BP 중에 상기 영여자 추출물을 용해시키고, 생성된 용액의 pH를 묽은 염산 BP를 사용하여 pH 3.5로 조절하고, 이어서 주사용 염화나트륨 BP를 사용하여 용적을 조절하고 충분히 혼합하였다. 용액을 투명 유리로 된 5㎖ 타입 Ⅰ 앰플 중에 충전시키고, 유리를 용해시킴으로써 공기의 상부 격자하에 봉입시키고, 이어서 120℃로 15분 이상 오토클래이브 시켜 살균하여 주사제를 얻었다. The above extracts were dissolved in an appropriate volume of sodium chloride BP for injection, and the pH of the resulting solution was adjusted to pH 3.5 with dilute hydrochloric acid BP, then the volume was adjusted with sodium chloride BP for injection and thoroughly mixed. The solution was filled in a 5 ml Type I ampoule made of clear glass, sealed under an upper grid of air by dissolving the glass, and then autoclaved at 120° C. for 15 minutes or more to sterilize to obtain an injection.
영여자 추출물 200 ㎍/㎖Young woman extract 200 ㎍/㎖
묽은 염산 BP pH 3.5로 될 때까지Dilute hydrochloric acid BP until pH 3.5
스테아르산 마그네슘 BP 최대 1㎖Magnesium Stearate BP up to 1ml
4. 환의 제조4. Preparation of pills
하기의 성분을 혼합한 후, 통산의 방법에 따라 1환 당 4g이 되도록 제조하였다. After mixing the following components, it was prepared so as to be 4 g per ring according to the general method.
영여자 추출물 0.5 g0.5 g of Young woman extract
유당 1.5 g1.5 g lactose
글리세린 1 g1 g of glycerin
자일리톨 0.5 g0.5 g of xylitol
5. 액제의 제조5. Preparation of liquid formulations
통상의 액제 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고, 레몬향을 적당량 가한 다음 하기의 성분을 혼합한 후 정제수를 가하여 전체를 100㎖로 조절하고 갈색병에 충진하여 멸균시켜 액제를 제조하였다. According to a conventional liquid preparation method, each component is added and dissolved in purified water, an appropriate amount of lemon flavor is added, and the following components are mixed, then purified water is added to adjust the total volume to 100 ml, filled in a brown bottle, and sterilized to prepare a liquid preparation did.
영여자 추출물 5 mgYoung woman extract 5 mg
이성화당 10 g10 g isomerized sugar
만니톨 5 g5 g of mannitol
정제수 적당량Appropriate amount of purified water
6. 산제의 제조6. Preparation of powder
하기의 성분을 혼합한 후, 기밀포에 충진하여 산제를 제조하였다. After mixing the following components, the powder was prepared by filling in an airtight cloth.
영여자 추출물 0.5 mg0.5 mg of young woman extract
유당 100 mg
탈크 10 mg
<제조예2> 건강기능 식품의 제조<Production Example 2> Preparation of health functional food
건강 식품의 제조manufacturing of health food
영여자 추출물 5 mgYoung woman extract 5 mg
비타민 혼합물 적량appropriate amount of vitamin mixture
비타민 A 아세테이트 70 ㎍70 μg vitamin A acetate
비타민 E 1 mg1 mg vitamin E
비타민 0.13 mgVitamin 0.13 mg
비타민 B2 0.15 mgVitamin B2 0.15 mg
비타민 B2 0.5 mg 0.5 mg of vitamin B2
비타민 B12 0.2 ㎍0.2 μg of vitamin B12
비타민 C 10 mg
비오틴 0.2 ㎍Biotin 0.2 μg
니콘틴산아미드 1.7 mgNicontinamide 1.7 mg
엽산 50 mg50 mg folic acid
판토텐산칼?? 0.5 mgPantothenic acid knife?? 0.5 mg
무기질혼합물 적량 Appropriate amount of inorganic mixture
황산제1철 1.75 mgferrous sulfate 1.75 mg
산화아연 0.82 mgZinc Oxide 0.82 mg
탄산마그네슘 25.3 mgMagnesium carbonate 25.3 mg
제1인산칼슘 15 mg
제2인산칼슘 55 mgDicalcium Phosphate 55 mg
구연산칼륨 90 mgPotassium citrate 90 mg
탄산칼슘 100 mg100 mg of calcium carbonate
염화마그네슘 24.8 mgMagnesium chloride 24.8 mg
상기 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 제조예로 혼합조성 하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강식품 조성물 제조에 사용할 수 있다. The composition ratio of the vitamin and mineral mixture was prepared by mixing components suitable for relatively healthy food as a preferred manufacturing example, but the mixing ratio may be arbitrarily modified, and after mixing the above ingredients according to a conventional health food manufacturing method, The granules can be prepared and used in the preparation of a health food composition according to a conventional method.
2. 건강음료의 제조2. Manufacture of health drinks
영여자 추출물 5.0 mgYoung woman extract 5.0 mg
구연산 1000 mgcitric acid 1000 mg
올리고당 100 g100 g of oligosaccharides
매실농축액 2gPlum Concentrate 2g
타우린 1g1 g of taurine
정제수를 가한 전체 900mlTotal 900ml with purified water
통상의 건강음료의 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간동안 85℃에서 교반 가열 후, 만들어진 용액을 여과하고 멸균된 2 리터(ℓ) 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관하여 본 발명에 따른 건강음료 조성물 제조에 사용하였다. 상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층이나 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라 그 배합비를 임의로 변형 실시하여도 무방하다. After mixing the above ingredients according to the usual manufacturing method of health drinks, after stirring and heating at 85°C for about 1 hour, the resulting solution is filtered and acquired in a sterilized 2 liter (ℓ) container, sealed and sterilized, and then stored in a refrigerator. Thus, it was used to prepare a health drink composition according to the present invention. Although the composition ratio is prepared by mixing ingredients suitable for relatively favorite beverages in a preferred embodiment, the mixing ratio may be arbitrarily modified according to regional and national preferences such as demand class, demanding country, and use.
상기에서 보는 바와 같이, 영여자 추출물은 전립선암 세포주에 대하여 세포독성을 나타내지 않음을 확인하였고, 전립선비대증 분자 진단 마커인 안드로겐 수용체 (androgenic receptor) 전립선 특이항원 (prostate specific antigen) 또는 5 알파-환원효소 (5-alpha reductase, 5-αR )의 발현을 억제시키는 효과를 가지므로, 전립선 질환 중 전립선비대증 또는 전립선암을 효과적으로 예방 및 치료 약학적 조성물 또는 건강기능식품 조성물로 유용하게 사용될 수 있다. As shown above, it was confirmed that the extract of Youngjae does not show cytotoxicity against prostate cancer cell lines, androgen receptor, prostate specific antigen, or 5 alpha-reductase, which is a molecular diagnostic marker for benign prostatic hyperplasia. (5-alpha reductase, 5-αR ) Since it has the effect of suppressing the expression, it can be effectively used as a pharmaceutical composition or a health functional food composition for effectively preventing and treating benign prostatic hyperplasia or prostate cancer among prostate diseases.
이제까지 본 발명에 바람직한 실시예 중심으로 살펴보았다. 본 발명이 속하는 기술 분야에서 통상의 지식을 가진 자는 본 발명이 본 발명의 본질적인 특성에서 벗어나지 않는 범위에서 변형된 형태로 구현될 수 있음을 이해할 수 있을 것이다. 그러므로 개시된 실시예들을 한정적인 관점이 아니라, 설명적인 관점에서 고려되어야 한다. 본 발명의 범위는 전술한 설명이 아니라 특허청구범위에 나타나 있으며, 그와 동등한 범위내에 있는 모든 차이점은 본 발명에 포함된 것으로 해석되어야 할 것이다.So far, the present invention has been focused on the preferred embodiment. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.
Claims (11)
A pharmaceutical composition for the prevention or treatment of prostate disease containing the extract of young woman as an active ingredient.
The pharmaceutical composition according to claim 1, wherein the extraction solvent of the extract is extracted with water, a C1 to C6 alcohol, or a mixture thereof.
The pharmaceutical composition according to claim 2, wherein the extraction solvent is a mixture of ethanol and water or ethanol.
The pharmaceutical composition according to claim 1, wherein the prostate disease is benign prostatic hyperplasia or prostate cancer.
The pharmaceutical composition according to claim 1, wherein the composition reduces the expression of 5-alpha reductase (5-αR) in prostate tissue.
The pharmaceutical composition according to claim 1, wherein the composition inhibits the expression of androgen receptor (AR) or prostate specific antigen (PSA).
The pharmaceutical composition according to claim 1, wherein the composition reduces libido or erectile dysfunction, which are side effects of prostate disease.
A health functional food composition for the prevention or improvement of prostate disease containing the extract of young woman as an active ingredient.
According to claim 8, wherein the extract is a health functional food composition, characterized in that the extraction solvent of the extract is extracted with water, C1 to C6 alcohol or a mixture thereof.
The health functional food composition according to claim 8, wherein the disease is benign prostatic hyperplasia or prostate cancer.
The health functional food composition according to claim 8, wherein the extract is prepared in any one formulation selected from beverages, pills, tablets, capsules, and powders.
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| KR101818741B1 (en) | 2008-09-16 | 2018-01-15 | 가니메드 파마슈티칼스 게엠베하 | Monoclonal antibodies for treatment of cancer |
| KR101908201B1 (en) | 2016-11-29 | 2018-10-15 | 김인숙 | Seasoned oil composition containing an ingredient of a flax seed and preparing process thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR101818741B1 (en) | 2008-09-16 | 2018-01-15 | 가니메드 파마슈티칼스 게엠베하 | Monoclonal antibodies for treatment of cancer |
| KR101908201B1 (en) | 2016-11-29 | 2018-10-15 | 김인숙 | Seasoned oil composition containing an ingredient of a flax seed and preparing process thereof |
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| Guo J 외. Clinical observation on ziyin tongbi decoction in treating benign prostatic hyperplasia. Chinese Journal of Integrated Traditional and Western Medicine. 2008, Vol. 28(12), pp. 1082-1085* |
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