KR20210074688A - The composition for treatment or prevention of benign prostatic hyperplasia comprising pretreated fermented soybean - Google Patents
The composition for treatment or prevention of benign prostatic hyperplasia comprising pretreated fermented soybean Download PDFInfo
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- KR20210074688A KR20210074688A KR1020190165658A KR20190165658A KR20210074688A KR 20210074688 A KR20210074688 A KR 20210074688A KR 1020190165658 A KR1020190165658 A KR 1020190165658A KR 20190165658 A KR20190165658 A KR 20190165658A KR 20210074688 A KR20210074688 A KR 20210074688A
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- Prior art keywords
- prostatic hyperplasia
- fermented
- present
- treatment
- benign prostatic
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Abstract
Description
본 발명은 전처리된 콩의 발효물을 포함하는 전립선비대증 치료용 조성물에 관한 것으로서, 보다 구체적으로는, 수침 후 건조시키는 과정을 반복수행한 콩의 발효물을 유효성분으로 포함하는 전립선비대증 예방, 치료 또는 개선용 조성물에 관한 것이다.The present invention relates to a composition for the treatment of benign prostatic hyperplasia comprising a fermented product of pretreated soybeans, and more specifically, to the prevention and treatment of benign prostatic hyperplasia, comprising, as an active ingredient, a fermented soybean obtained by repeatedly drying after water immersion. Or it relates to a composition for improvement.
전립선은 샘 조직과 섬유근조직으로 구성된 남성의 부속생식선으로 전립선에 관련한 질환으로는 전립선염, 전립선비대증 및 전립선암이 있고, 이 중 50세 이상의 남성에게 가장 흔하게 나타나는 질환은 전립선비대증(benign prostatic hyperplasia. BPH)이다. 전립선비대증은 남자 노인들의 삶의 질을 저해하는 대표적인 질환이다. 전립선비대증은 요도 주위에 위치한 전립선의 부피가 증가하여 요도를 압박하고 이로 인해 빈뇨, 절박뇨 등의 방광 저장 증상과 지연뇨, 단절뇨, 배뇨시 힘을 주어야 하는 현상 등 방광의 배출 장애를 나타내는 증상을 통칭한 것이라 할 수 있다.The prostate is an accessory gonad in men composed of glandular tissue and fibromuscular tissue. Diseases related to the prostate include prostatitis, benign prostatic hyperplasia, and prostate cancer. BPH). BPH is a representative disease that impairs the quality of life of the elderly male. BPH is characterized by an increase in the volume of the prostate located around the urethra, which compresses the urethra, resulting in bladder storage symptoms such as frequent urination and urgency, and symptoms indicating bladder discharge disorders such as delayed urination, interrupted urination, and the need to exert force during urination. It can be called a generic name.
전립선비대증의 약물 치료제는 크게 알파차단제와 안드로겐억제제(5알파환원효소 억제제)로 분류된다. 알파차단제는 전립선요도의 압력과 긴장을 낮추어주는 약물로서 시중에서 판매되는 알파차단제로는 테라조신(terazosin), 독사조신(doxazosin), 탐스로신(tamsulosin), 알푸조신(alfuzosin) 등이 있다. 그러나 알파차단제는 전립선의 비대된 현상을 줄여주는 효과는 없으므로 일시적인 치료방법일 뿐이며, 어지럼증, 무기력증, 두통, 시야 장애 등의 부작용이 보고되었다. 안드로겐억제제는 전립선의 비대 과정에서 핵심적인 역할을 수행하는 5알파 환원효소를 억제하여 전립선의 비대를 막아주는 약물이다. 5알파환원효소 억제제로서 시중에는 피나스테리드(finasteride)와 두타스테리드(dutasteride)가 판매되고 있다. 그러나 이들 약물은 복용 후 효과가 나타날 때까지 시간이 오래 걸리며, 성기능 관련 부작용이 있다.Drug treatment for benign prostatic hyperplasia is largely divided into alpha blockers and androgen inhibitors (5-alpha reductase inhibitors). Alpha blockers are drugs that lower the pressure and tension of the prostatic urethra, and commercially available alpha blockers include terazosin, doxazosin, tamsulosin, and alfuzosin. However, alpha blockers have no effect on reducing the enlarged prostate, so they are only a temporary treatment method, and side effects such as dizziness, lethargy, headache, and visual field disturbance have been reported. Androgen inhibitors are drugs that prevent enlargement of the prostate by inhibiting 5-alpha reductase, which plays a key role in the process of enlargement of the prostate. As 5-alpha reductase inhibitors, finasteride and dutasteride are commercially available. However, these drugs take a long time to take effect after taking them, and there are side effects related to sexual function.
이와 같이, 현재까지 개발 중이거나 시판되고 있는 약물은 대부분 부작용이 많이 나타나므로, 부작용이 적으면서 효능이 우수한 새로운 치료제의 개발이 요구되고 있다.As described above, since most of the drugs being developed or marketed to date have many side effects, there is a demand for the development of a new therapeutic agent with few side effects and excellent efficacy.
한편, 콩은 대표적인 식물성 단백질(35 ~ 45%) 식품일 뿐만 아니라 지방질(10 ~ 15%), 탄수화물(10%) 및 무기질(5%)의 균형적인 급원으로 특히 한국인에게는 두부, 장류, 두유, 콩기름 등의 식품의 형태로 소비되고 있다. 콩은 영양학적 측면뿐만 아니라 사포닌, 이소플라본, 레시틴과 같은 생리활성 물질에 관한 연구가 활발해지면서 기능성 식품의 재료로서 각광받고 있다. 특히, 특히 대두 이소플라본은 항산화활성 뿐만 아니라 유방암 및 전립선암 등에 효과적인 것으로 알려져 있다. On the other hand, soybeans are not only a representative plant protein (35 ~ 45%) food but also a balanced source of fat (10 ~ 15%), carbohydrates (10%) and minerals (5%). It is consumed in the form of foods such as soybean oil. Soybeans are in the spotlight as a functional food ingredient as research on physiologically active substances such as saponins, isoflavones, and lecithins, as well as nutritional aspects, is active. In particular, soy isoflavones are known to be effective for breast and prostate cancer as well as antioxidant activity.
따라서, 콩의 효능을 증진시키기 위해, 콩 품종, 재배 환경, 발아, 가공 처리 비배당체 형태로의 전환 기술(한국등록특허 10-1161231호 참고)에 대한 연구가 시도되고 있으나, 아직 체계화된 방법이 존재하지 않고, 세밀하고 복잡한 과정을 거쳐야 하는 문제점이 있다.Therefore, in order to enhance the efficacy of soybeans, research on soybean varieties, cultivation environment, germination, processing and conversion technology to non-glycoside form (refer to Korean Patent No. 10-1161231) has been attempted, but a systematic method is not yet available. There is a problem that does not exist, and must go through a detailed and complicated process.
상기와 같은 문제점을 해결하기 위하여, 본 발명자들은 콩을 발효시키 전에 수침하여 건조시키는 전처리 과정을 수회 수행하는 간단한 방법을 통해, 이소플라본의 함량의 변화는 없음에도 불구하고 전립선비대증의 개선 및 치료 효능이 유의적으로 증진되는 것을 확인함으로써, 본 발명을 완성하게 되었다.In order to solve the above problems, the present inventors, through a simple method of performing a pre-treatment process of water immersion and drying several times before fermenting soybeans, despite no change in the content of isoflavones, improvement of prostatic hyperplasia and therapeutic efficacy By confirming that this significantly improved, the present invention was completed.
이에, 본 발명은 수침 후 건조시킨 콩의 발효물을 유효성분으로 포함하는 전립선비대증 예방 또는 치료용 약학적 조성물을 제공하는 것을 목적으로 한다.Accordingly, an object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of benign prostatic hyperplasia comprising a fermented soybean dried after water immersion as an active ingredient.
또한, 본 발명은 수침 후 건조시킨 콩의 발효물을 유효성분으로 포함하는 전립선비대증 예방 또는 개선용 건강기능식품 조성물을 제공하는 것을 다른 목적으로 한다.In addition, another object of the present invention is to provide a health functional food composition for preventing or improving prostatic hyperplasia comprising a fermented soybean dried after water immersion as an active ingredient.
그러나 본 발명이 이루고자 하는 기술적 과제는 이상에서 언급한 과제에 제한되지 않으며, 언급되지 않은 또 다른 과제들은 아래의 기재로부터 당업자에게 명확하게 이해될 수 있을 것이다.However, the technical problem to be achieved by the present invention is not limited to the above-mentioned problems, and other problems not mentioned will be clearly understood by those skilled in the art from the following description.
상기 목적을 달성하기 위하여, 본 발명은 수침 후 건조시킨 콩의 발효물을 유효성분으로 포함하는 전립선비대증 치료용 약학적 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for the treatment of benign prostatic hyperplasia comprising a fermented soybean dried after water immersion as an active ingredient.
본 발명의 일 구현예로, 상기 수침 후 건조과정을 2 내지 3회 반복수행할 수 있다.In one embodiment of the present invention, the drying process after the water immersion may be repeated 2-3 times.
본 발명의 다른 구현예로, 상기 조성물은 안드로겐 수용체(Androgen receptor), PCNA(proliferation cell nulear antigen), PSA (prostate specific antigen) 및 Bcl-xL로 이루어진 군으로부터 선택되는 하나 이상의 mRNA 발현을 억제할 수 있다. In another embodiment of the present invention, the composition can inhibit the expression of one or more mRNAs selected from the group consisting of androgen receptor (Androgen receptor), PCNA (proliferation cell nulear antigen), PSA (prostate specific antigen) and Bcl-xL. have.
또한, 본 발명은 수침 후 건조시킨 콩의 발효물을 유효성분으로 포함하는 전립선비대증 예방 또는 개선용 건강기능식품 조성물을 제공한다.In addition, the present invention provides a health functional food composition for preventing or improving benign prostatic hyperplasia comprising a fermented soybean dried after water immersion as an active ingredient.
본 발명의 일 구현예로, 상기 식품은 콩제품, 유제품, 발효유, 발효식품 및 발효음료로 구성된 군에서 선택되는 어느 하나의 형태일 수 있다.In one embodiment of the present invention, the food may be in any one form selected from the group consisting of soy products, dairy products, fermented milk, fermented food and fermented beverage.
본 발명에 따라 전처리 수행된 콩의 발효물은 간단한 전처리 공정만을 수행함으로써 기존 콩의 발효물에 비하여 전립선비대증의 개선 및 치료효과가 유의적으로 향상되는바, 전립선비대증을 예방, 개선 또는 치료할 수 있다는 다양한 약학적 조성물 또는 건강기능식품에 유용하게 이용될 수 있다.The fermented soybean pre-treated according to the present invention significantly improves the improvement and therapeutic effect of prostatic hyperplasia compared to the conventional fermented soybean by performing only a simple pre-treatment process, and thus it is possible to prevent, improve or treat prostatic hyperplasia. It can be usefully used in various pharmaceutical compositions or health functional foods.
도 1은 본 발명에 따라 전처리된 콩 발효물의 세포독성을 확인한 결과를 나타낸 것이다.
도 2는 본 발명에 따라 전처리된 콩 발효물의 안드로겐 수용체(Androgen receptor) 발현 억제능 결과를 나타낸 것이다.
도 3은 본 발명에 따라 전처리된 콩 발효물의 PCNA(proliferation cell nulear antigen) 발현 억제능 결과를 나타낸 것이다.
도 4는 본 발명에 따라 전처리된 콩 발효물의 PSA (prostate specific antigen) 발현 억제능 결과를 나타낸 것이다.
도 5는 본 발명에 따라 전처리된 콩 발효물의 Bcl-xL 발현 억제능 결과를 나타낸 것이다.
도 6a는 전처리에 처리 유무에 따른 콩의 발효물 내 이소플라본 함량 변화를 확인한 1차(2017) 결과를 나타낸 것이다.
도 6b는 전처리에 처리 유무에 따른 콩의 발효물 내 이소플라본 함량 변화를 확인한 2차(2018) 결과를 나타낸 것이다.1 shows the result of confirming the cytotoxicity of the soybean fermented product pretreated according to the present invention.
Figure 2 shows the results of androgen receptor (Androgen receptor) expression inhibition of the pre-treated soybean fermented according to the present invention.
Figure 3 shows the results of inhibiting the expression of PCNA (proliferation cell nulear antigen) expression of pre-treated soybean fermented according to the present invention.
Figure 4 shows the PSA (prostate specific antigen) expression inhibition results of the pre-treated soybean fermented according to the present invention.
Figure 5 shows the results of the inhibition of Bcl-xL expression of the soybean fermented product pretreated according to the present invention.
Figure 6a shows the first (2017) result confirming the change in the isoflavone content in the fermented soybean according to the presence or absence of pretreatment.
Figure 6b shows the second (2018) result confirming the change in the isoflavone content in the fermented soybean according to the presence or absence of pretreatment.
본 발명자들은 콩을 발효시키 전에 수침하여 건조시키는 전처리 과정을 수행하는 경우, 전립선비대증의 개선 및 치료 효능이 유의적으로 증진됨을 확인함으로써, 본 발명을 완성하였다.The present inventors have completed the present invention by confirming that the improvement and therapeutic efficacy of prostatic hyperplasia are significantly enhanced when the pretreatment process of drying the soybeans by water soaking before fermentation is performed.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 수침 후 건조시킨 콩의 발효물을 유효성분으로 포함하는 전립선비대증 예방 또는 치료용 약학적 조성물을 제공한다.The present invention provides a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising a fermented soybean dried after water immersion as an active ingredient.
본 발명에서 사용되는 용어, "예방"이란 본 발명에 따른 약학적 조성물의 투여에 의해 전립선비대증을 억제시키거나 발병을 지연시키는 모든 행위를 의미한다.As used herein, the term “prevention” refers to any action that suppresses or delays the onset of BPH by administration of the pharmaceutical composition according to the present invention.
본 발명에서 사용되는 용어, "치료"란 본 발명에 따른 약학적 조성물의 투여에 의해 전립선비대증에 대한 증세가 호전되거나 이롭게 변경되는 모든 행위를 의미한다.As used herein, the term “treatment” refers to any action in which the symptoms of benign prostatic hyperplasia are improved or beneficially changed by administration of the pharmaceutical composition according to the present invention.
본 발명에서 콩을 전처리하는 과정은 콩을 물에 담궈서 수분을 제공하는 수침과정과 수침된 콩을 말리는 건조과정으로 이루어질 수 있다. 상기 수침과정은 콩 3배~4배의 물을 가하여 15~20℃에서 2-5시간 동안 수침할 수 있으며, 바람직하게는 16~17℃에서 2-3시간 동안 수침할 수 있으나, 콩에 충분한 수분을 제공한다면 수침의 조건이 특별히 한정되는 것은 아니다. 또한, 건조과정은 35~45℃에서 2일 내지 4일간 건조시킬 수 있으며, 바람직하게는 40~41℃에서 3일간 건조시킬 수 있으나, 수침된 콩에서 수분을 모두 제거한다면 건조의 조건이 특별히 한정되는 것은 아니다.In the present invention, the process of pre-treating the beans may include a water immersion process in which the beans are immersed in water to provide moisture, and a drying process in which the soaked beans are dried. The water immersion process can be immersed in water for 2-5 hours at 15-20 ℃ by adding 3 to 4 times the water of the beans, and preferably, it can be immersed for 2-3 hours at 16-17 ℃, but it is sufficient for soybeans If moisture is provided, the conditions of water immersion are not particularly limited. In addition, the drying process can be dried for 2 to 4 days at 35 ~ 45 ℃, preferably dried for 3 days at 40 ~ 41 ℃, but if all moisture is removed from the soaked soybeans, the drying conditions are particularly limited. it's not going to be
본 발명에 있어서, 상기 수침 및 건조 공정은 수회 반복하여 실시하는 것이 바람직하며, 더 바람직하게는 2 내지 3회, 가장 바람직하게는 3회 반복하여 실시할 수 있다.In the present invention, the water immersion and drying process is preferably repeated several times, more preferably 2 to 3 times, most preferably 3 times may be repeated.
본 발명에 있어서, 발효는 당 업계에서 통상적으로 수행되는 방식을 사용할 수 있으며, 예컨대 콩을 수침시켜 불린 후 삶아서 35℃ 내지 45℃에서 2일 내지 4일간 숙성시키는 단계를 통해 수행될 수 있다.In the present invention, the fermentation may use a method commonly performed in the art, for example, it may be carried out through the step of soaking and soaking soybeans and then boiling and aging at 35 ° C. to 45 ° C. for 2 to 4 days.
본 발명자들은 구체적인 실시예를 통해 수침과정 및 건조과정으로 이루어진 전처리공정을 수행한 콩의 발효물의 경우 전립선비대증 치료 효능이 유의적으로 증진됨을 실험적으로 확인하였다.The present inventors experimentally confirmed that the treatment efficacy of benign prostatic hyperplasia is significantly improved in the case of a fermented soybean that has been subjected to a pretreatment process consisting of a water immersion process and a drying process through specific examples.
보다 구체적으로, 본 발명의 일 실시예에서는, 본 발명에 따른 콩의 발효물의 전립선비대증 치료효능을 확인한 결과, 전처리를 수행하지 않은 콩의 발효물과 비교할 때, 전처리를 수행한 경우에 안드로겐 수용체(Androgen receptor), PCNA(proliferation cell nulear antigen), PSA (prostate specific antigen) 및 Bcl-xL의 mRNA 발현을 유의적으로 억제시키는 것을 확인하였다(실시예 4 내지 7 참조).More specifically, in one embodiment of the present invention, as a result of confirming the prostatic hyperplasia treatment efficacy of the fermented soybean according to the present invention, when compared with the fermented soybean that is not pre-treated, androgen receptor ( Androgen receptor), it was confirmed that the mRNA expression of PCNA (proliferation cell nulear antigen), PSA (prostate specific antigen) and Bcl-xL was significantly inhibited (see Examples 4 to 7).
본 발명의 또 다른 실시예에서는, 전처리에 따른 콩의 발효물의 이소플라본 함량을 비교한 결과, 전처리 과정을 수행에 따른 총 이소플라본 함량의 변화에는 차이가 없음을 확인하였다(실시예 8 참조).In another embodiment of the present invention, as a result of comparing the isoflavone content of fermented soybeans according to the pretreatment, it was confirmed that there was no difference in the change in the total isoflavone content according to the pretreatment process (see Example 8).
상기 실시예의 결과를 통해, 본 발명에 따라 전처리 수행된 콩의 발효물은 이소플라본의 함량의 변화가 없음에도 불구하고, 전립선비대증에 대한 치료 효능을 유의적으로 우수하다는 것을 알 수 있다.Through the results of the above examples, it can be seen that the fermented soybean pretreated according to the present invention has significantly superior therapeutic efficacy for BPH, despite no change in the content of isoflavones.
본 발명에 따른 약학적 조성물은 약학적으로 허용 가능한 담체를 더 포함할 수 있다. 상기 약학적으로 허용 가능한 담체는 제제시에 통상적으로 이용되는 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 사이클로덱스트린, 덱스트로즈 용액, 말토덱스트린 용액, 글리세롤, 에탄올, 리포좀 등을 포함하지만 이에 한정되지 않으며, 필요에 따라 항산화제, 완충액 등 다른 통상의 첨가제를 더 포함할 수 있다. 또한, 희석제, 분산제, 계면활성제, 결합제, 윤활제 등을 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다. 적합한 약학적으로 허용되는 담체 및 제제화에 관해서는 레밍턴의 문헌에 개시되어 있는 방법을 이용하여 각 성분에 따라 바람직하게 제제화할 수 있다. 본 발명의 약학적 조성물은 제형에 특별한 제한은 없으나 주사제, 흡입제, 피부 외용제 등으로 제제화할 수 있다. The pharmaceutical composition according to the present invention may further include a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is commonly used in formulation, and includes, but is not limited to, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, dextrose solution, maltodextrin solution, glycerol, ethanol, liposome, and the like. It does not, and may further include other conventional additives such as antioxidants and buffers, if necessary. In addition, diluents, dispersants, surfactants, binders, lubricants, etc. may be additionally added to form an injectable formulation such as an aqueous solution, suspension, emulsion, etc., pills, capsules, granules or tablets. Regarding suitable pharmaceutically acceptable carriers and formulations, formulations can be preferably made according to each component using the method disclosed in Remington's literature. The pharmaceutical composition of the present invention is not particularly limited in the formulation, but may be formulated as an injection, an inhalant, or an external preparation for skin.
본 발명의 약학적 조성물은 목적하는 방법에 따라 경구 투여하거나 비경구투여(예를 들어, 정맥 내, 피하, 복강 내 또는 국소에 적용)할 수 있으며, 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 시간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다.The pharmaceutical composition of the present invention may be administered orally or parenterally (eg, intravenously, subcutaneously, intraperitoneally or topically) according to a desired method, and the dosage may vary depending on the condition and weight of the patient, and the disease. Although it varies depending on the degree, drug form, administration route and time, it may be appropriately selected by those skilled in the art.
본 발명의 약학적 조성물은 약학적으로 유효한 양으로 투여한다. 본 발명에 있어서 "약학적으로 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 배출비율, 치료기간, 동시 사용되는 약물을 포함한 요소 및 기타 의학 분야에 잘 얄려진 요소에 따라 결정될 수 있다. 본 발명에 다른 약학적 조성물은 개별 치료제로 투여하거나 다른 치료제와 병용하여 투여될 수 있고 종래 의 치료제와는 순차적 또는 동시에 투여될 수 있으며, 단일 또는 다중 투여될 수 있다. 상기한 요소들을 모두 고려하여 부작용 없이 최소한의 양으로 최대 효과를 얻을 수 있는 양을 투여하는 것이 중요하며, 이는 당업자에 의해 용이하게 결정될 수 있다.The pharmaceutical composition of 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 is determined by the type, severity of the patient's disease, the activity of the drug, Sensitivity to the drug, administration time, administration route and excretion rate, treatment period, factors including concurrent drugs and other factors well known in the medical field may be determined. The pharmaceutical composition according to 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 conventional therapeutic agents, and may be administered single or multiple. In consideration of all of the above factors, it is important to administer an amount that can obtain the maximum effect with a minimum amount without side effects, which can be easily determined by those skilled in the art.
구체적으로 본 발명의 약학적 조성물의 유효량은 환자의 연령, 성별, 상태, 체중, 체내에 활성 성분의 흡수도, 불활성율 및 배설속도, 질병종류, 병용되는 약물에 따라 달라질 수 있으며, 일반적으로는 체중 1kg 당 0.001 내지 150mg, 바람직하게는 0.01 내지 100mg을 매일 또는 격일 투여하거나, 1일 1 내지 3회로 나누어 투여할 수 있다. 그러나 투여 경로, 비만의 중증도, 성별, 체중, 연령 등에 따라서 증감 될 수 있으므로 상기 투여량이 어떠한 방법으로도 본 발명의 범위를 한정하는 것은 아니다.Specifically, the effective amount of the pharmaceutical composition of the present invention may vary depending on the patient's age, sex, condition, weight, absorption of the active ingredient into the body, inactivation rate and excretion rate, disease type, and drugs used in combination, in general 0.001 to 150 mg per 1 kg of body weight, preferably 0.01 to 100 mg, may be administered daily or every other day, or divided into 1 to 3 times a day. However, the dosage may be increased or decreased according to the route of administration, the severity of obesity, sex, weight, age, etc., and thus the dosage is not intended to limit the scope of the present invention in any way.
본 발명의 다른 양태로서, 본 발명은 상기 약학적 조성물을 개체에 투여하는 단계를 포함하는 전립선비대증의 예방 또는 치료방법을 제공한다. As another aspect of the present invention, the present invention provides a method for preventing or treating benign prostatic hyperplasia comprising administering the pharmaceutical composition to an individual.
본 발명에서 "개체"란 질병의 치료를 필요로 하는 대상을 의미하고, 보다 구체적으로는, 인간 또는 비-인간인 영장류, 생쥐(mouse), 개, 고양이, 말, 및 소 등의 포유류를 의미한다.In the present invention, "individual" refers to a subject in need of treatment for a disease, and more specifically, refers to mammals such as human or non-human primates, mice, dogs, cats, horses, and cattle. do.
본 발명의 다른 양태로서, 본 발명은 수침 후 건조시킨 콩의 발효물을 유효성분으로 포함하는 전립선비대증 예방 또는 개선용 건강기능식품 조성물을 제공한다.As another aspect of the present invention, the present invention provides a health functional food composition for preventing or improving benign prostatic hyperplasia comprising a fermented soybean dried after water immersion as an active ingredient.
본 발명에서 사용되는 용어, "개선"이란 치료되는 상태와 관련된 파라미터, 예를 들면 증상의 정도를 적어도 감소시키는 모든 행위를 의미한다. 이때 상기 식품 조성물은 전립선비대증의 예방 또는 개선을 위하여 해당 질환의 발병 단계 이전 또는 발병 후, 치료를 위한 약제와 동시에 또는 별개로서 사용될 수 있다.As used herein, the term “improvement” refers to any action that at least reduces a parameter related to the condition being treated, for example, the severity of symptoms. In this case, the food composition may be used simultaneously with or separately from a drug for treatment before or after the onset of the disease for the prevention or improvement of benign prostatic hyperplasia.
본 발명에서 '식품'은 콩제품 (두유, 두부, 콩고기, 콩스택, 콩과자), 유제품 (우유, 치즈), 발효유 (액상 요구르트, 호상 요구르트), 발효식품 (김치류, 장류, 피클), 발효 음료 등의 형태가 될 수 있으나 이에 제한되지는 않는다.In the present invention, 'food' means soybean products (soymilk, tofu, soybean meat, soybean stack, soybean cake), dairy products (milk, cheese), fermented milk (liquid yogurt, yoghurt), fermented food (kimchi, soy sauce, pickles), It may be in the form of a fermented beverage, but is not limited thereto.
상기 본 발명에 더 포함될 수 있는 첨가제로는, 천연 탄수화물, 향미제, 영양제, 비타민, 광물(전해질), 풍미제(합성 풍미제, 천연 풍미제 등), 착색제, 충진제, 팩트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 산화 방지제, 글리세린, 알코올, 탄산화제 및 과육으로 이루어진 군으로부터 선택된 1종 이상의 성분을 사용할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토오스, 수크로오스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 상기 향미제로서 천연 향미제(타우마틴, 스테비아추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 외에 본 발명에 따른 조성물은 여러 가지 영양제, 비타민, 광물 (전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제, 팩트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 상기 담체, 부형제, 희석제 및 첨가제의 구체적인 예로는 이에 한정하는 것은 아니나, 락토오스, 덱스트로즈, 수크로오스, 솔비톨, 만니톨, 에리스리톨, 전분, 아카시아 고무, 인산칼슘, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 미세결정성 셀룰로즈, 폴리비닐키롤리돈, 셀룰로즈, 폴리비닐피로리돈, 메틸셀룰로즈, 물, 설탕시럽, 메틸 하이드록시 벤조에이트, 프로필하이드록시 벤조에이트, 활석, 스테아트산 마그네슘 및 미네랄 오일로 이루어진 그룹으로부터 선택된 1종 이상이 사용되는 것이 바람직하다.Additives that may be further included in the present invention include natural carbohydrates, flavoring agents, nutrients, vitamins, minerals (electrolytes), flavoring agents (synthetic flavoring agents, natural flavoring agents, etc.), coloring agents, fillers, lactic acid and salts thereof, At least one component selected from the group consisting of alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, antioxidants, glycerin, alcohols, carbonation agents, and pulp may be used. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agent, natural flavoring agents (taumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The composition according to the invention comprises various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and flavoring agents such as natural flavoring agents, coloring agents and thickening agents, facic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners , pH adjusting agent, stabilizer, preservative, glycerin, alcohol, carbonation agent used in carbonated beverage, etc. These components can be used independently or in combination. Specific examples of the carrier, excipient, diluent and additive is, but not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, erythritol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium phosphate, calcium silicate, microcrystalline cellulose, polyvinylkyrolidone, At least one selected from the group consisting of cellulose, polyvinylpyrrolidone, methylcellulose, water, sugar syrup, methyl hydroxy benzoate, propyl hydroxy benzoate, talc, magnesium stearate and mineral oil is preferably used .
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시한다. 그러나 하기의 실시예는 본 발명을 보다 쉽게 이해하기 위하여 제공되는 것일 뿐, 하기 실시예에 의해 본 발명의 내용이 한정되는 것은 아니다.Hereinafter, preferred examples are presented to help the understanding of the present invention. However, the following examples are only provided for easier understanding of the present invention, and the contents of the present invention are not limited by the following examples.
[실시예][Example]
실시예 1. 실험준비 및 실험방법Example 1. Experimental preparation and experimental method
1-1. 시료 준비1-1. Sample preparation
발효콩을 80℃에서 3시간 동안 30% 에탄올을 이용하여 추출하였다. 추출물을 여과시킨 뒤, Rotary vaccum evaporator을 사용하여 환류 냉각하여 농축하였다. 농축한 추출물을 동결건조하여 실험에 이용하였다.Fermented soybeans were extracted using 30% ethanol at 80° C. for 3 hours. After the extract was filtered, it was concentrated by reflux cooling using a rotary vacuum evaporator. The concentrated extract was freeze-dried and used in the experiment.
1-2. BPH-1 / RWPE-1의 세포배양 1-2. Cell culture of BPH-1 / RWPE-1
전립선 비대 상피세포인 BPH-1 세포주는 20% FBS, penicillin (100 U/ml), 그리고 100μg/ml의 streptomycin을 첨가한 RPMI 1640 배지로 37℃, 5% CO2 incubator에서 배양하였다. 정상 전립선 상피세포인 RWPE-1 세포주는 recombinant human Epidermal Growth Factor (rhEGF)와 Bovine Pituitary Extract (BPE)로 구성된 첨가제, penicillin (100 U/ml), 그리고 100μg/ml의 streptomycin을 첨가한 K-SFM 배지로 37℃, 5% CO2 incubator에서 배양하였다.The BPH-1 cell line, an enlarged prostate epithelial cell, was cultured in RPMI 1640 medium supplemented with 20% FBS, penicillin (100 U/ml), and 100 μg/ml streptomycin in an incubator at 37° C., 5% CO 2 . The RWPE-1 cell line, which is a normal prostate epithelial cell, contains K-SFM medium supplemented with recombinant human Epidermal Growth Factor (rhEGF) and Bovine Pituitary Extract (BPE), penicillin (100 U/ml), and streptomycin at 100 μg/ml. 37 ℃, 5% CO 2 Incubated in an incubator.
1-3. MTT assay1-3. MTT assay
BPH-1 및 RWPE-1 세포를 well 당 2×105의 밀도로 96-well plate에 seeding 하고, 다양한 농도의 실험소재를 세포에 처리하였다. 24시간 이후 3-(4, 5-Dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) 용액을 처리하였으며, 처리 4시간 이후 상층 부유물을 제거하고, dimethyl sulfoxide(DMSO)을 이용하여 비수용성 formazan product을 용해하였다. Cell viability는 Epoch® microvolume spectrophotometer system (BioTek Instruments Inc., Winooski, VT, USA)을 이용하여 570 nm 파장대에서 측정되었다. DHT를 처리한 RWPE-1 세포를 이용하여 발효콩의 androgen-sensitive 전립선 세포의 proliferation에 대한 효능을 확인하고자 RWPE-1 세포주에 대하여 MTT assay를 진행하였다.BPH-1 and RWPE-1 cells were seeded in a 96-well plate at a density of 2×10 5 per well, and various concentrations of test material were treated on the cells. After 24 hours, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) solution was treated, and after 4 hours, the supernatant was removed, and dimethyl sulfoxide (DMSO) was used. The non-aqueous formazan product was dissolved. Cell viability was measured at a wavelength of 570 nm using an Epoch® microvolume spectrophotometer system (BioTek Instruments Inc., Winooski, VT, USA). In order to confirm the efficacy on proliferation of androgen-sensitive prostate cells of fermented soybeans using DHT-treated RWPE-1 cells, MTT assay was performed on the RWPE-1 cell line.
1-4. Real-Time PCR1-4. Real-Time PCR
표적 분자의 유전자를 증폭하기 위해 올리고핵산 primer 염기서열은 NCBI data base에 보고된 것을 기초로 Primer3 software를 이용하여 primer를 디자인하였다. 일반 PCR 반응을 통해 디자인한 primer들의 특이성을 확인하고, annealing temperature와 MgCl2 농도를 변화시켜 PCR 조건을 최적화하였다. 이어 증폭된 DNA가 목적하는 산물인지를 확인하기 위하여 PCR product의 DNA sequencing을 실시하였다. mRNA는 Syber-green RT-PCR을 통하여 정량하고, standard curve는 total RNA (200 pg~320 pg)의 5배 희석액을 사용하여 정립하였다. 해리곡선 (dissociation curve)은 non-specific amplification이 없음을 확인한 후, Syber-green assay을 수행하였으며, 모든 결과는 housekeeping gene인 GAPDH로 표준화하였다 Real-time PCR 결과는 Sequence Detection System software을 사용하여 분석하였다.To amplify the gene of the target molecule, the primer was designed using Primer3 software based on the nucleotide sequence of the oligonucleic acid primer reported in the NCBI data base. The specificity of the designed primers was confirmed through a general PCR reaction, and the PCR conditions were optimized by changing the annealing temperature and MgCl 2 concentration. Next, DNA sequencing of the PCR product was performed to confirm whether the amplified DNA was the desired product. mRNA was quantified through Syber-green RT-PCR, and a standard curve was established using a 5-fold dilution of total RNA (200 pg~320 pg). After confirming that there was no non-specific amplification for the dissociation curve, Syber-green assay was performed, and all results were standardized to GAPDH, a housekeeping gene. Real-time PCR results were analyzed using Sequence Detection System software. .
실시예 2. 콩의 발효물 제조Example 2. Preparation of fermented products of soybeans
2-1. 콩의 준비2-1. preparation of beans
본 연구에 사용된 콩 품종은 '대풍'은 백운콩과 신팔달콩2호의 교배조합으로 2002년에 품종등록 되었으며, 주로 두부 및 장류용으로 이용된다. '대풍' 콩의 재배는 고령지농업연구소 진부포장(해발고도 540m)에서 2016 ~ 2018년에 재배·정선하여 준비하였다.The soybean variety used in this study, 'Daepoong', was registered in 2002 as a hybrid combination of Baegun soybean and Shinpaldalkong No. 2, and is mainly used for tofu and soy sauce. 'Daepung' beans were cultivated and selected from 2016 ~ 2018 in Jinbu Field (540m above sea level) at the Goryeongji Agricultural Research Institute.
2-2. 전처리 과정2-2. pretreatment process
상기 실시예 1-1에서 준비한 콩에 대하여 하기의 조건으로 수침 및 건조 과정을 수행하였다.Water soaking and drying processes were performed on the beans prepared in Example 1-1 under the following conditions.
1회 : 16~17℃의 물에 2시간 담근 후 40~41℃의 건조기에서 72시간 충분히 건조1 time: After soaking in water at 16~17℃ for 2 hours, dry thoroughly for 72 hours in a dryer at 40~41℃
2회 : 1회 건조된 콩을 다시 16~17℃의 물에 2시간 담근 후 40~41℃의 건조기에서 72시간 충분히 건조2nd time: After soaking the dried beans in water at 16~17℃ for 2 hours, dry them sufficiently in a dryer at 40~41℃ for 72 hours.
3회 : 2회 건조처리된 콩을 다시 16~17℃의 물에 2시간 담근 후 40~41℃의 건조기에서 72시간 충분히 건조3 times: Soybeans that have been dried twice are immersed in water at 16~17℃ for 2 hours and then thoroughly dried in a dryer at 40~41℃ for 72 hours.
상기 전처리된 콩은 i) 1회 전처리만 수행한 그룹 ii) 1차 및 2차 전처리만 수행한 그룹 iii) 3차까지 모두 수행한 그룹으로 분류하여 실험을 진행하였다.The pre-treated soybeans were classified into i) a group in which only one pre-treatment was performed, ii) a group in which only the first and second pre-treatments were performed, iii) a group in which all of the three pretreatments were performed, and the experiment was carried out.
2-3. 콩 발효2-3. soybean fermentation
상기 실시예 2-2에서 준비된 콩(무처리, 수침 및 건조 처리 1회, 2회, 3회)을 16~17℃의 물에 3시간 30분 담근 후 끓어 넘치지 않게 3시간 30분간 삶고 삶아진 콩은 발효실로 옮겼다. 채반에 짚을 깔고 그 위에 면보를 덮은 후 면보 위에 삶은 콩은 펼쳐 놓고 면보를 덮어 42℃에서 72시간 발효하여 최종 콩 발효물을 얻었다.Soybeans prepared in Example 2-2 (untreated, immersed in water and dried once, twice, and three times) were soaked in water at 16-17° C. for 3 hours and 30 minutes, then boiled and boiled for 3 hours and 30 minutes without boiling over. Soybeans were transferred to the fermentation chamber. Straw was laid on a sieve, a cotton cloth was placed on it, and then boiled soybeans were spread out on the cotton cloth, covered with a cotton cloth, and fermented at 42°C for 72 hours to obtain the final fermented soybean product.
실시예 3. 전처리된 콩의 발효물의 세포독성 확인Example 3. Confirmation of cytotoxicity of fermented soybeans pretreated
정상 전립선 상피세포인 RWPE-1 세포주에 대하여 상기 실시예 2에서 준비된 콩(무처리, 수침 및 건조 처리 1회, 2회, 3회)의 발효물을 농도별(15.625, 31.25, 62.5, 125, 250, 500, 1000μg/ml)로 처리하여 RWPE-1 세포 생존률에 미치는 영향을 확인해보았다.For the RWPE-1 cell line, which is a normal prostate epithelial cell, the fermented product of the soybean (untreated, water-immersed and dried once, twice, and three times) prepared in Example 2 was subjected to concentration (15.625, 31.25, 62.5, 125, 250, 500, and 1000 μg/ml) to check the effect on RWPE-1 cell viability.
그 결과, 도 1에 나타낸 바와 같이, 1회, 2회 및 3회 전처리를 수행한 콩의 발효물 모두 세포독성을 나타내지 않는 것을 확인할 수 있었다. As a result, as shown in FIG. 1, it was confirmed that none of the fermented soybeans subjected to pretreatment once, twice and three times exhibited cytotoxicity.
실시예 4. 전처리된 콩의 발효물의 안드로겐 수용체(Androgen receptor) 발현 억제능 확인Example 4. Androgen receptor (Androgen receptor) expression inhibitory ability of the fermented soybean pretreated
전립선 비대 상피세포인 BPH-1 세포주에 실시예 2를 통해 준비된 각각의 콩 발효물(무처리, 수침 및 건조 처리 1회, 2회, 3회)을 처리한 후, 전립선 표적분자인 androgen recepor(AR)의 mRNA 발현 변화를 RT-PCR 실험을 통해 확인하였다.After treating the BPH-1 cell line, which is an enlarged prostate epithelial cell, each soybean fermented product prepared in Example 2 (untreated, water immersed and dried 1, 2, 3 times), androgen recepor ( AR) mRNA expression changes were confirmed through RT-PCR experiments.
그 결과, 도 2에 나타낸 바와 같이, 전처리 과정을 수행한 콩의 발효물을 처리한 경우에서 안드로겐 수용체(Androgen receptor)의 mRNA 발현이 억제되는 것을 확인할 수 있었다. 보다 구체적으로, 전처리를 수행하지 않은 경우(control) 대비 1회 처리 48%, 2회 처리 61%, 3회 처리 5% 수준으로 발현이 억제되었다. 특히, 1회 및 2회 전처리를 수행한 경우보다 3회 전처리를 수행한 경우에 안드로겐 수용체(Androgen receptor)의 mRNA 발현이 유의적으로 억제되었다. As a result, as shown in FIG. 2 , it was confirmed that the mRNA expression of androgen receptor was suppressed when the fermented soybean subjected to the pretreatment process was treated. More specifically, expression was suppressed at a level of 48% of treatment once, 61% of treatment twice, and 5% of
실시예 5. 전처리된 콩의 발효물의 PCNA(proliferation cell nulear antigen) 발현 억제능 확인Example 5. Confirmation of PCNA (proliferation cell nulear antigen) expression inhibitory ability of fermented soybeans pretreated
전립선 비대 상피세포인 BPH-1 세포주에 실시예 2를 통해 준비된 각각의 콩 발효물(무처리, 수침 및 건조 처리 1회, 2회, 3회)을 처리한 후, 세포 과증식 대표 마커인 PCNA(proliferation cell nulear antigen)의 mRNA 발현 변화를 RT-PCR 실험을 통해 확인하였다.After treating each of the soybean fermented products prepared in Example 2 (untreated, water soaked and dried once, twice, and three times) to the BPH-1 cell line, which is an enlarged prostate epithelial cell, PCNA (a representative marker of cell hyperproliferation) ( The change in mRNA expression of proliferation cell null antigen) was confirmed through RT-PCR experiment.
그 결과, 도 3에 나타낸 바와 같이, 전처리 과정을 수행한 콩의 발효물을 처리한 경우에서 PCNA(proliferation cell nulear antigen)의 mRNA 발현이 억제되는 것을 확인할 수 있었다. 보다 구체적으로, 전처리를 수행하지 않은 경우(no treatment) 대비, 1회 처리 94%, 2회 처리 86%, 3회 처리 79% 수준으로 발현이 억제되었다.As a result, as shown in FIG. 3 , it was confirmed that the mRNA expression of PCNA (proliferation cell nulear antigen) was suppressed when the fermented soybean that had been pre-treated was treated. More specifically, expression was suppressed at a level of 94% treatment once, 86% treatment twice, 79
실시예 6. 전처리된 콩의 발효물의 PSA (prostate specific antigen) 발현 억제능 확인Example 6. Confirmation of inhibiting ability of PSA (prostate specific antigen) expression of fermented soybeans pretreated
전립선 비대 상피세포인 BPH-1 세포주에 실시예 2를 통해 준비된 각각의 콩 발효물(무처리, 수침 및 건조 처리 1회, 2회, 3회)을 처리한 후, 전립선 암 뿐만 아니라 전립선 비대 질활의 진단에 이용되는 전립선 특이 항원인 PSA (prostate specific antigen)의 mRNA 발현 변화를 RT-PCR 실험을 통해 확인하였다.BPH-1 cell line, which is an enlarged prostate epithelial cell, was treated with each of the soybean fermented products prepared in Example 2 (untreated, water immersed and dried once, twice, and three times), and not only prostate cancer but also prostate cancer Changes in the mRNA expression of PSA (prostate specific antigen), a prostate-specific antigen used for diagnosis, were confirmed through RT-PCR experiments.
그 결과, 도 4에 나타낸 바와 같이, 전처리 과정을 수행한 콩의 발효물을 처리한 경우에서 PSA (prostate specific antigen)의 mRNA 발현이 억제되는 것을 확인할 수 있었다. 보다 구체적으로, 전처리를 수행하지 않은 경우(no treatment)와 대비할 때, 1회 처리한 경우에는 215%로 증가한 반면, 2회 처리 경우에는 91%, 3회 처리 경우에는 9% 수준으로 발현이 억제되었다. 특히, 1회 및 2회 전처리를 수행한 경우보다 3회 전처리를 수행한 경우에 PSA (prostate specific antigen)의 mRNA 발현이 유의적으로 억제되는 것을 확인할 수 있었다. As a result, as shown in FIG. 4 , it was confirmed that the mRNA expression of PSA (prostate specific antigen) was suppressed when the fermented soybean that had been subjected to the pretreatment process was treated. More specifically, when compared to the case where no pretreatment was performed (no treatment), the expression was suppressed to a level of 215% in the case of one treatment, whereas 91% in the case of the second treatment and 9% in the case of the third treatment. became In particular, it was confirmed that the mRNA expression of PSA (prostate specific antigen) was significantly suppressed when the pretreatment was performed 3 times compared to the case where the pretreatment was performed 1 and 2 times.
실시예 7. 전처리된 콩의 발효물의 Bcl-xL 발현 억제능 확인Example 7. Confirmation of Bcl-xL expression inhibitory ability of fermented soybeans pretreated
Bcl-xL은 세포자멸사를 촉진시키는 Bax의 작용을 억제함으로써 세포 생존율을 촉진시키는 역할을 한다. 이에, 전립선 비대 상피세포인 BPH-1 세포주에 실시예 2를 통해 준비된 각각의 콩 발효물(무처리, 수침 및 건조 처리 1회, 2회, 3회)을 처리한 후, 전립선 특이 항원인 Bcl-xL의 mRNA 발현 변화를 RT-PCR 실험을 통해 확인하였다.Bcl-xL promotes cell viability by inhibiting the action of Bax that promotes apoptosis. Accordingly, after treating each of the soybean fermented products prepared in Example 2 (untreated, water immersed and dried once, twice, and three times) to the BPH-1 cell line, which is an enlarged prostate epithelial cell, the prostate-specific antigen Bcl -xL mRNA expression change was confirmed through RT-PCR experiment.
그 결과, 도 5에 나타낸 바와 같이, 전처리 과정을 수행한 콩의 발효물을 처리한 경우에서 Bcl-xL의 mRNA 발현이 억제되는 것을 확인할 수 있었다. 보다 구체적으로, 전처리를 수행하지 않은 경우(no treatment)와 대비할 때, 1회 처리한 경우에는 168%로 증가한 반면, 2회 처리 경우에는 67%, 3회 처리 경우에는 37% 수준으로 발현이 억제되었다. 특히, 1회 및 2회 전처리를 수행한 경우보다 3회 전처리를 수행한 경우에 Bcl-xL의 mRNA 발현이 유의적으로 억제되는 것을 확인할 수 있었다. As a result, as shown in FIG. 5 , it was confirmed that the mRNA expression of Bcl-xL was suppressed when the fermented soybean which had been subjected to the pretreatment process was treated. More specifically, when compared to the case where no pretreatment was performed (no treatment), the expression was suppressed to a level of 168% in the case of one treatment, 67% in the case of the second treatment, and 37% in the case of the third treatment. became In particular, it was confirmed that the mRNA expression of Bcl-xL was significantly suppressed when the pretreatment was performed three times than when the pretreatment was performed once and twice.
실시예 8. 전처리에 따른 이소플라본 함량 비교Example 8. Comparison of isoflavone content according to pretreatment
전처리된 콩의 발효물의 전립선비대증 효과 증진이 상기 발효물 내 이소플라본 함량의 증가에 따른 것인지 확인하기 위해, 전처리에 따른 이소플라본 함량을 비교하였다.In order to determine whether the enhancement of the prostatic hyperplasia effect of the fermented soybean pre-treated is due to an increase in the isoflavone content in the fermented product, the isoflavone content according to the pre-treatment was compared.
보다 구체적으로, 발효된 콩을 동결건조 후 각각의 시료 1g을 취하여 70% 에탄올 25ml를 넣고 30분간 교반하였고 이를 여과지(Whatman No.2)를 이용하여 필터링하였다. 얻어진 용액을 50ml로 정량하고 syringe filter로 다시 필터링하여 준비하였다. 이소플라본 분석기기는 UPLC(Waters, Acquity UPLC I-Class, USA)로 분석하였다. 컬럼은 BEH C18(2.0×50mm, 17㎛)을 이용하였으며, 시료 주입량은 1㎕였다. 유속은 0.2㎖/min이었고 260nm 파장에서 검출하였다. 이동상 A는 0.1% acetic acid(in water), 이동상 B는 acetonitrile를 이용하였다.More specifically, after freeze-drying fermented soybeans, 1 g of each sample was taken, 25 ml of 70% ethanol was added, stirred for 30 minutes, and filtered using filter paper (Whatman No. 2). The obtained solution was quantified to 50 ml and prepared by filtering again with a syringe filter. The isoflavone analyzer was analyzed by UPLC (Waters, Acquity UPLC I-Class, USA). As a column, BEH C18 (2.0×50 mm, 17 μm) was used, and the sample injection amount was 1 μl. The flow rate was 0.2 ml/min and detection was performed at a wavelength of 260 nm. Mobile phase A was 0.1% acetic acid (in water), and mobile phase B was acetonitrile.
상기 실험방법에 따라 상기 실시예 2를 통해 준비된 콩 (무처리, 건조처리 1회, 2회, 3회)에 대하여 이소플라본 성분을 분석하였고, 실험은 2번(2017년, 2018년) 수행하였으며, 분석 결과를 도 6a 및 도 6b에 나타내었다.According to the experimental method, isoflavone components were analyzed for the soybeans prepared in Example 2 (untreated, dried 1 time, 2 times, 3 times), and the experiment was performed twice (2017, 2018). , the analysis results are shown in FIGS. 6A and 6B .
상기 도 6a 및 도 6b에 나타낸 바와 같이, 전처리 과정을 수행에 따른 총 이소플라본 함량의 변화에는 차이가 없음을 확인할 수 있었다.As shown in FIGS. 6A and 6B, it was confirmed that there was no difference in the change in the total isoflavone content according to the pretreatment process.
상기 결과로부터, 전처리 과정 수행에 따른 콩 발효물의 전립선비대증 효과 증진이 이소플라본 함량 증가에 의한 것은 아님을 알 수 있었다.From the above results, it could be seen that the enhancement of the prostatic hyperplasia effect of the soybean fermented product according to the pre-treatment was not due to the increase in the isoflavone content.
전술한 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야의 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다. 그러므로 이상에서 기술한 실시예들은 모든 면에서 예시적인 것이며 한정적이 아닌 것으로 이해해야만 한다.The description of the present invention described above is for illustration, and those of ordinary skill in the art to which the present invention pertains can understand that it can be easily modified into other specific forms without changing the technical spirit or essential features of the present invention. will be. Therefore, it should be understood that the embodiments described above are illustrative in all respects and not restrictive.
Claims (5)
A pharmaceutical composition for the treatment of benign prostatic hyperplasia comprising, as an active ingredient, a fermented soybean dried after water immersion.
The pharmaceutical composition according to claim 1, wherein the drying process is repeated 2-3 times after the water immersion.
The method according to claim 1, wherein the composition inhibits the expression of one or more mRNAs selected from the group consisting of androgen receptor (Androgen receptor), PCNA (proliferation cell nulear antigen), PSA (prostate specific antigen), and Bcl-xL. which is a pharmaceutical composition.
A health functional food composition for improving prostatic hyperplasia, comprising, as an active ingredient, a fermented soybean dried after water immersion.
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