WO2023090542A1 - Pharmaceutical composition containing hot water extract of fruit of vaccinium bracteatum thunb. as active ingredient for preventing or treating benign prostatic hyperplasia - Google Patents
Pharmaceutical composition containing hot water extract of fruit of vaccinium bracteatum thunb. as active ingredient for preventing or treating benign prostatic hyperplasia Download PDFInfo
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- WO2023090542A1 WO2023090542A1 PCT/KR2022/004445 KR2022004445W WO2023090542A1 WO 2023090542 A1 WO2023090542 A1 WO 2023090542A1 KR 2022004445 W KR2022004445 W KR 2022004445W WO 2023090542 A1 WO2023090542 A1 WO 2023090542A1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/45—Ericaceae or Vacciniaceae (Heath or Blueberry family), e.g. blueberry, cranberry or bilberry
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P13/00—Drugs for disorders of the urinary system
- A61P13/08—Drugs for disorders of the urinary system of the prostate
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- the present invention relates to a composition for preventing and treating benign prostatic hyperplasia containing an extract of the fruit of moss japonica as an active ingredient. More specifically, it relates to a pharmaceutical composition for preventing or treating benign prostatic hyperplasia that can be safely used without toxicity and side effects by using the fruit of Vaccinium bracteatum Thunb., which is a natural raw material.
- Benign prostatic hyperplasia is a disease in which the volume of the prostate around the urethra increases for various reasons due to various causes, compresses the urethra, and as a result, symptoms such as decreased urine output occur.
- urinary frequency (more than 8 times/day), nocturia, urgency (strong and sudden need to urinate), urgency (difficulty urinating), retardation (delayed urine excretion), and intermittent urination (urine flow is interrupted). It is defined as a complaint of lower urinary tract symptoms, which collectively refers to symptoms indicating bladder emptying disorders, such as urinary incontinence and the need to exert force during urination. It is known that the cause of prostate enlargement is related to the male hormones testosterone and dihydrotestosterone (DHT).
- DHT dihydrotestosterone
- Alpha-blockers are drugs that lower pressure and tension in the prostate and urethra, and include terazosin, doxazosin, tamsulosin, and alfuzosin. Side effects such as disability have been reported.
- Androgen inhibitors are drugs that prevent enlargement of the prostate by inhibiting 5-alpha-reductase, and include finasteride and dutasteride. Side effects related to sexual function have been reported.
- Vaccinium bracteatum Thunb. is a dicotyledonous angiosperm plant of the order Rhododendron, which grows in coastal mountains. The height is 1-3m, and the twigs are grayish brown to gray and almost hairless. The leaves are alternate, thick, elliptical or oblong, with a thick leathery texture. There are fine sawtooth on the edge, and there is a small dot under the backside. The flower blooms in June and is reddish-white, and about 10 bell-shaped flowers hang on the raceme hanging down and bracts remain. Fruits are berries, round, covered with white powder, about 6mm in diameter, ripe in October, and edible.
- composition for preventing or treating prostatic hyperplasia that can be safely used without toxicity and side effects by extracting an active ingredient from the fruit of the moss tree, which is a natural raw material.
- the present invention includes a fruit extract of Vaccinium Bracteatum Thunb. as an active ingredient, and the extract is an extract available from any one selected from water, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Characterized in that It provides a functional food composition for preventing or treating benign prostatic hyperplasia or a pharmaceutical composition for preventing or improving benign prostatic hyperplasia.
- the pharmaceutical composition for preventing or treating benign prostatic hyperplasia of the present invention may be provided by containing 0.1 to 2000 mg/kg/body weight/day of the extract of the moss fruit or 0.01 to 99.9% by weight of the composition.
- the pharmaceutical composition may be formulated in any one form selected from powders, granules, tablets, capsules, suspensions, emulsions, syrups, transdermal preparations, suppositories, or sterile injectable solutions.
- the functional food composition may be prepared in any one form selected from tablets and capsules, soft capsules, granules, and liquids.
- composition containing the hot-water extract of the fruit of the present invention has an activity of reducing testosterone-induced prostate (LNCaP) or prostate weight, suppressing 5-alpha-reductase-2 production, and reducing tissue polyps.
- the present invention is a composition for preventing or treating prostatic hyperplasia using a mosae tree fruit extract as an active ingredient, using mosae tree, a domestic natural resource, and safely preventing or treating prostatic hyperplasia without toxicity or side effects, and as a health functional food composition.
- Figure 1 shows the powder and liquid phase of the extract of the fruit of Mosaenamu prepared in the present invention.
- Figure 2 is a graph showing the survival rate of the prostate epithelial cell line (LNCaP) for the moss extract.
- Figure 3 shows the effect on the contents of prostate-specific enzyme (5 ⁇ -reductase 2), dihydrotestosterone (DHT), and prostate-specific antigen (PSA) according to the treatment of the robin fruit extract in the prostate epithelial cell line (LNCaP) by testosterone. It is a picture.
- prostate-specific enzyme (5 ⁇ -reductase 2), dihydrotestosterone (DHT), and prostate-specific antigen (PSA) according to the treatment of the robin fruit extract in the prostate epithelial cell line (LNCaP) by testosterone. It is a picture.
- Figure 4 is a picture showing the androgen receptor (AR) protein expression according to testosterone-induced prostatic epithelial cell line (LNCaP) treatment with moss berry extract.
- AR androgen receptor
- Figure 5 is a picture showing the effect of inhibiting prostate tissue hypertrophy according to the treatment of the moss berry extract in a testosterone-induced benign prostatic rat model.
- Figure 6 is a picture showing the effect of inhibiting the content of 5-alpha reductase in prostate tissue according to the treatment of the moss berry extract in a rat model of prostatic hyperplasia induced by testosterone.
- FIG. 7 is a diagram showing a cross-sectional staining photograph of prostate tissue according to the treatment of a moss fruit extract in a testosterone-induced benign prostatic rat model.
- the present invention relates to a composition for preventing or treating benign prostatic hyperplasia containing an extract of the fruit of moss japonica as an active ingredient, and the configuration and effects of the present invention will be described in more detail through specific examples and comparative examples.
- Mosae tree fruit extract was extracted by adding 50 L of distilled water to 5 kg of dried Moss tree fruit by removing moisture, and heating at 100 ° C. for 3 hours. Filtered, reduced pressure and concentrated. The concentrated hot-water extract was frozen and dried at -40 ° C to 40 ° C for 72 hours using a freeze dryer.
- Figure 1 shows the powder and liquid phase of the extract of the fruit of the moss tree prepared in the present invention.
- Figure 2 is a graph showing the survival rate of the prostate epithelial cell line (LNCaP) for the moss extract.
- LNCaP cells Korea Cell Line Bank 21740
- FBS fetal bovine serum
- penicillin and streptomycin 37°C and 5% CO 2 conditions.
- the cultured cells were dispensed in a 96-well plate at a concentration of 1x10 5 cell/well, cultured, and after 12 hours, the extract of the fruit of the moss tree prepared in the present invention was treated at a concentration of 0, 10, 50, and 100 ⁇ g/ml, respectively. .
- 10 ⁇ L of EZCytox solution was added to each well, and toxicity was confirmed by measuring absorbance at 450 nm using a plate reader. As shown in FIG. 2, it was confirmed that the extract of the fruit of the moss tree had no cytotoxicity up to 50 ⁇ g/ml.
- Figure 3 shows the effect on the contents of prostate-specific enzyme (5 ⁇ -reductase 2), dihydrotestosterone (DHT), and prostate-specific antigen (PSA) according to the treatment of the robin fruit extract in the prostate epithelial cell line (LNCaP) by testosterone. It is a picture.
- prostate-specific enzyme (5 ⁇ -reductase 2), dihydrotestosterone (DHT), and prostate-specific antigen (PSA) according to the treatment of the robin fruit extract in the prostate epithelial cell line (LNCaP) by testosterone. It is a picture.
- LNCaP cells were dispensed in a 6-well plate at a concentration of 1x10 6 cells/well and cultured at 37°C.
- the cells were treated with 1 ⁇ M of testosterone, and simultaneously treated with the prepared hot-water extract at concentrations of 0, 5, 10, and 50 ⁇ g/ml, and then harvested after 24 hours of culture.
- a positive control BPH
- 10 ⁇ M of Finasteride a treatment for prostatic hyperplasia, was treated with 1 ⁇ M of testosterone instead of the extract.
- the untreated group was used as a negative control group.
- Phosphate-buffered saline pH7.4 (Gibco, USA) was added to the cells harvested after culture, and homogenization was performed with a Microcentrifuge Sample Pestle. The cell homogenate was centrifuged at 4°C and 5,000 rpm for 5 minutes (Mikro 200R, Hettich, USA) to separate the supernatant, and 5-alpha reductase, dihydrotestosterone and prostate were separated using each ELISA KIT (CUSABIO, USA). Specific antigens were measured.
- Figure 4 is a picture showing the results of testosterone-induced androgen receptor (AR) protein expression in prostate epithelial cell line (LNCaP) according to the treatment of moss berry extract.
- Prostate epithelial cells (LNCaP) were treated with testosterone and moss fruit extract, and the protein expression of androgen receptor (AR) was confirmed to confirm whether the moss fruit extract was effective in prostatic hyperplasia.
- LNCaP cells were dispensed in a 6-well plate at a concentration of 1x10 6 and cultured at 37°C.
- the cells were treated with 1 ⁇ M of testosterone, and simultaneously treated with the prepared hot-water extract at concentrations of 0, 5, 10, and 50 ⁇ g/ml, and then harvested after 24 hours of culture.
- a positive control group BPH
- a control group Fea
- 10 ⁇ M of Finasteride a treatment for prostatic hyperplasia, was treated with 1 ⁇ M of testosterone instead of the extract.
- the untreated group was used as a negative control group.
- RIPA buffer 50 mM Tris-HCl, pH 8.0, with 150 mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate, with a protease inhibitor cocktail
- RIPA buffer 50 mM Tris-HCl, pH 8.0, with 150 mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate, with a protease inhibitor cocktail
- the separated protein was quantified using a protein assay kit (Bio-Rad protein assay kit, Bio-Rad, USA), and the quantified protein was subjected to SDS-PAGE on a 10% polyacrylamide gel, and the protein was transferred to a PVDF membrane. .
- the PVDF membrane is reacted with 5% skim milk for about 1 hour to remove non-specific binding proteins on the surface, and the primary antibody, anti-AR antibody or anti- ⁇ -actin antibody, is used for one day at 4 ° C. reacted Thereafter, the secondary antibody was treated at room temperature for 1 hour, and the level of AR protein expression was confirmed using an ECL kit (Thermo scientific, USA). As shown in FIG. 4, it was confirmed that the AR protein expression inhibitory effect was remarkably superior to the control group in the cell group treated with 5, 10, or 50 ⁇ g/ml of the moss fruit extract.
- Figure 5 is a picture showing the effect of inhibiting prostate tissue hypertrophy according to treatment with hot water extract of moss tree fruit in a rat model of prostatic hyperplasia induced by testosterone.
- TP testosterone propionate
- the hot-water extract of the fruit of mosae tree was orally administered at 50 and 100 mg/kg for 4 weeks, and finasteride (FI), a 5 ⁇ -reductase inhibitor used for the treatment of benign prostatic hyperplasia, was orally administered at a dose of 5 mg/kg as a positive control group.
- finasteride a 5 ⁇ -reductase inhibitor used for the treatment of benign prostatic hyperplasia
- each group normal group; Sham, prostatic hyperplasia induced group (negative control group); BPH, prostatic hyperplasia induced group + finasteride administration group (positive control group); FI, benign prostatic hyperplasia group + moss fruit extract 50mg/kg administration group; VBF50, benign prostatic hyperplasia induction group + mosae tree fruit extract 100mg/kg administration group; prostates were removed from VBF100 and weighed The left prostate tissue was removed and 5 ⁇ -reductase was measured in the tissue, and the prostate growth inhibitory effect (%) was calculated according to the following equation.
- Prostatic hyperplasia inhibitory effect 100 - ⁇ (prostate weight in the seed-treated group or positive control group - normal prostate weight) * 100 / (negative control prostate weight - normal prostate weight) ⁇
- Table 1 shows a table of prostate tissue weights according to the treatment of moss berry extract in a testosterone-induced benign prostatic rat model.
- the prostate size was 0.30 ⁇ 0.01 in the control group, 29.7% compared to 0.18 ⁇ 0.02 in the normal group. has increased.
- Prostate hyperplasia growth inhibitory effect was 73.56% in the positive control group, and 20.84% and 28.75% respectively in the group administered with 50 or 100mg/kg of moss fruit, showing inhibitory effects in a concentration-dependent manner.
- Alpha-reductase measurement Figure 6 is a diagram showing the inhibitory effect of 5-alpha-reductase content in prostate tissue according to the treatment of the moss fruit extract in a rat model of prostatic hyperplasia induced by testosterone.
- the prostate tissue of the sacrificed rat was washed once with physiological saline and homogenized for 1 minute with a Wise Stir homogenizer (Daihan Scientific, Korea) after adding 4 times phosphate-buffered saline pH7.4 (Gibco, USA).
- Prostate tissue homogenate was centrifuged at 4°C and 5,000 rpm for 5 minutes (Mikro 200R, Hettich, USA) to separate the supernatant. measured.
- FIG. 7 is a diagram showing a cross-sectional staining photograph of prostate tissue according to the treatment of a moss fruit extract in a testosterone-induced benign prostatic rat model.
- the right prostate tissues of the sacrificed rats were fixed in 10% neutral formalin, paraffin-embedded and sectioned, and stained with Hematoxylin-Eosin. Histological changes in the epithelial cells and stroma of the prostate were observed under an optical microscope.
- compositions or health foods for the prevention or treatment of prostatic hyperplasia using the fruit extract of moss tree as an active ingredient may be prepared in the form of tablets and capsules, soft capsules, granules, and liquids.
- the pharmaceutical formulation or health food may be prepared as a beverage additive.
- the pharmaceutical formulation or health food for preventing or treating prostatic hyperplasia is a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol such that the composition containing the mosae tree fruit extract as an active ingredient is contained in an amount of 0.01 to 99.9% by weight , It can be prepared by formulating into a transdermal preparation, a suppository or a sterile injectable solution.
- the pharmaceutical composition in the case of a sterile injection solution, can be prepared by mixing 99.9 to 0.01% by weight of purified water or glucose with 0.01 to 99.9% by weight of the pharmaceutical composition, and in the case of capsules, freeze-drying the pharmaceutical composition to 0.01 to 99.9% by weight %, and can be prepared by mixing 99.9 to 0.01% by weight of vitamins and calcium.
- the daily dosage of the pharmaceutical composition prepared above is provided in an amount of 10 to 2000 mg/kg body weight of the extract, and a pharmaceutical preparation for preventing or treating enlargement of the prostate or health containing 0.01 to 99.9% by weight of the pharmaceutical composition. can be made into food.
- the present invention is a composition for preventing or inhibiting prostatic hyperplasia, which contains an extract of moss fruit as an active ingredient, and is useful for preventing or inhibiting prostatic hyperplasia in testosterone-induced prostate cells (LNCaP) and an enlarged prostate rat model, so it has industrial applicability. .
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Abstract
The present invention relates to a composition for preventing or treating benign prostatic hyperplasia, the composition containing, as an active ingredient, an extract of the fruit of Vaccinium bracteatum thunb. The composition inhibits the in vitro generation of 5 alpha reductase, dihydrotestosterone, and androgen receptor (AR). In a benign prostatic hyperplasia rat model induced by testosterone, the composition inhibits prostate tissue enlargement, inhibits 5-alpha reductase generation in prostate hypertrophy tissue, relieves enlarged epithelial cells, and inhibits polyps, thus exhibiting an excellent benign prostatic hyperplasia prevention or inhibition effect in animal models.
Description
본 발명은 모새나무 열매 추출물을 유효성분으로 함유하는 전립선 비대증 예방 및 치료용 조성물에 관한 것이다. 보다 구체적으로는 천연원료인 모새나무(Vaccinium bracteatum Thunb.) 열매를 이용하여 독성 및 부작용 없이 안전하게 사용될 수 있는 전립선 비대증 예방 또는 치료용 약학 조성물에 관한 것이다.The present invention relates to a composition for preventing and treating benign prostatic hyperplasia containing an extract of the fruit of moss japonica as an active ingredient. More specifically, it relates to a pharmaceutical composition for preventing or treating benign prostatic hyperplasia that can be safely used without toxicity and side effects by using the fruit of Vaccinium bracteatum Thunb., which is a natural raw material.
전립선 비대증(Benign prostatic hyperplasia, BPH)은 전립선 질환의 하나로 다양한 원인으로 인해 요도 주위의 전립선의 부피가 증가하여 요도를 압박하고 이로 인해 소변의 배출 감소 등의 증세가 발생하는 질환이다. Benign prostatic hyperplasia (BPH) is a disease in which the volume of the prostate around the urethra increases for various reasons due to various causes, compresses the urethra, and as a result, symptoms such as decreased urine output occur.
최근에는 남성에서 빈뇨(8회 이상/일), 야간뇨, 긴박뇨(강하고 갑작스런 요의), 절박뇨(소변을 참기 어려움), 지연뇨(소변의 배출이 지연됨), 단절뇨(소변의 흐름이 끊김), 배뇨시 힘을 주어야 하는 현상 등의 방광 배출 장애를 나타내는 증상을 통칭한 하부 요로증상의 호소로 정의되고 있다. 전립선 비대의 원인으로 남성호르몬인 테스토스테론(testosterone)과 디하이드로테스토스테론(dihydrotestosterone, DHT)이 관련되어 있다고 알려져 있다.Recently, in men, urinary frequency (more than 8 times/day), nocturia, urgency (strong and sudden need to urinate), urgency (difficulty urinating), retardation (delayed urine excretion), and intermittent urination (urine flow is interrupted). It is defined as a complaint of lower urinary tract symptoms, which collectively refers to symptoms indicating bladder emptying disorders, such as urinary incontinence and the need to exert force during urination. It is known that the cause of prostate enlargement is related to the male hormones testosterone and dihydrotestosterone (DHT).
전립선 비대증의 약물 치료제는 크게 알파차단제와 안드로겐 억제제(5알파-환원효소 억제제)로 분류된다. 알파차단제는 전립선요도의 압력과 긴장을 낮추어주는 약물로, 테라조신(terazosin), 독사조신(doxazosin), 탐스로신(tamsulosin), 알푸조신(alfuzosin) 등이 있는데, 어지럼증, 무기력증, 두통, 시야 장애 등의 부작용이 보고된바 있다. 안드로겐억제제는 5알파-환원효소를 억제하여 전립선의 비대를 막아주는 약물로, 피나스테라이드(finasteride)와 두타스테리드(dutasteride) 등이 있는데, 성기능 관련 부작용이 보고된 바 있다.Drug treatment for BPH is largely classified into alpha blockers and androgen inhibitors (5 alpha-reductase inhibitors). Alpha-blockers are drugs that lower pressure and tension in the prostate and urethra, and include terazosin, doxazosin, tamsulosin, and alfuzosin. Side effects such as disability have been reported. Androgen inhibitors are drugs that prevent enlargement of the prostate by inhibiting 5-alpha-reductase, and include finasteride and dutasteride. Side effects related to sexual function have been reported.
한편 모새나무(Vaccinium bracteatum Thunb.) 는 진달래목의 쌍떡잎 속씨식물로 해변의 산지에서 자란다. 높이가 1∼3m로, 작은가지는 회갈색에서 회색이고 거의 털이 없다. 잎은 어긋나고 두꺼우며 타원형 또는 긴 타원형으로 두꺼운 가죽같은 질감을 갖는다. 가장자리에 잔 톱니가 있으며 뒷면 밑에 작은 선점이 있으며, 꽃은 6월에 피고 홍백색이 돌며 밑으로 처지는 총상꽃차례에 종처럼 생긴 꽃이 10여 개씩 달리고 포가남아 있다. 열매는 장과로 둥글고 흰가루로 덮이며 지름 6mm 정도로 10월에 익으며 먹을 수 있다.On the other hand, Vaccinium bracteatum Thunb. is a dicotyledonous angiosperm plant of the order Rhododendron, which grows in coastal mountains. The height is 1-3m, and the twigs are grayish brown to gray and almost hairless. The leaves are alternate, thick, elliptical or oblong, with a thick leathery texture. There are fine sawtooth on the edge, and there is a small dot under the backside. The flower blooms in June and is reddish-white, and about 10 bell-shaped flowers hang on the raceme hanging down and bracts remain. Fruits are berries, round, covered with white powder, about 6mm in diameter, ripe in October, and edible.
현대인의 생활수준이 향상됨에 따라 부작용이 거의 없고 자연에서 채취가 가능한 천연자원에 대해서 관심이 높아지고 있다. 특히 전통적으로 알려진 모새나무와 같은 천연물의 효능을 연구하여 부작용이 거의 없고 질병의 예방과 회복에 도움이 되는 물질을 개발하기 위해 노력하고 있으나, 아직까지는 모새나무에 대한 약학적 가치나 생리활성 기능성에 대한 관련정보가 부족한 실정이다. As the standard of living of modern people improves, interest in natural resources that have little side effects and can be collected from nature is increasing. In particular, efforts are being made to develop a substance that has few side effects and is helpful in preventing and recovering from diseases by studying the efficacy of natural products such as traditionally known mosae tree. Relevant information is lacking.
천연원료인 모새나무 열매로부터 유효 성분을 추출하여 독성 및 부작용 없이 안전하게 사용될 수 있는 전립선 비대증 예방 또는 치료용 조성물을 제공한다.Provided is a composition for preventing or treating prostatic hyperplasia that can be safely used without toxicity and side effects by extracting an active ingredient from the fruit of the moss tree, which is a natural raw material.
본 발명은 모새나무(Vaccinium Bracteatum Thunb.) 열매 추출물을 유효성분으로 포함하며, 상기 추출물은 물 또는 탄소수 1 내지 4의 저급 알코올 또는 이들의 혼합 용매 중에서 선택되는 어느 하나로부터 가용한 추출물인 것을 특징으로 하는 전립선 비대증 예방 또는 치료용 약학 조성물 또는 전립선 비대증 예방 또는 개선용 기능성 식품조성물을 제공한다.The present invention includes a fruit extract of Vaccinium Bracteatum Thunb. as an active ingredient, and the extract is an extract available from any one selected from water, lower alcohol having 1 to 4 carbon atoms, or a mixed solvent thereof. Characterized in that It provides a functional food composition for preventing or treating benign prostatic hyperplasia or a pharmaceutical composition for preventing or improving benign prostatic hyperplasia.
본 발명의 전립선 비대증 예방 또는 치료용 약학 조성물은 모새나무 열매 추출물이 0.1 내지 2000 mg/kg/체중/1일의 양으로 또는 상기 조성물을 0.01 내지 99.9중량%로 함유하여 제공될 수 있다. 상기 약학 조성물은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 경피제, 좌제 또는 멸균 주사용 액 중에서 선택되는 어느 하나의 형태로 제형화될 수 있다. 기능성 식품조성물은 정제 및 캡슐제, 연질 캡슐제, 과립제, 액제 중에서 선택되는 어느 하나의 형태로 제조될 수 있다.The pharmaceutical composition for preventing or treating benign prostatic hyperplasia of the present invention may be provided by containing 0.1 to 2000 mg/kg/body weight/day of the extract of the moss fruit or 0.01 to 99.9% by weight of the composition. The pharmaceutical composition may be formulated in any one form selected from powders, granules, tablets, capsules, suspensions, emulsions, syrups, transdermal preparations, suppositories, or sterile injectable solutions. The functional food composition may be prepared in any one form selected from tablets and capsules, soft capsules, granules, and liquids.
본 발명의 모새나무 열매 열수 추출물을 포함하는 조성물은 테스토스테론을 유도한 전립선(LNCaP) 또는 전립선 무게 감소, 5알파환원효소-2 생성 억제 및 조직 폴립을 감소시키는 활성을 가진다. The composition containing the hot-water extract of the fruit of the present invention has an activity of reducing testosterone-induced prostate (LNCaP) or prostate weight, suppressing 5-alpha-reductase-2 production, and reducing tissue polyps.
본 발명은 모새나무 열매 추출물을 유효성분으로 하는 전립선 비대증 예방 또는 치료용 조성물로써 국내 천연자원인 모새나무를 자원으로 이용하고 독성이나 부작용 없이 안전하게 전립선 비대증의 예방 또는 치료용 조성물 및 건강 기능성 식품조성물로서 사용될 수 있다.The present invention is a composition for preventing or treating prostatic hyperplasia using a mosae tree fruit extract as an active ingredient, using mosae tree, a domestic natural resource, and safely preventing or treating prostatic hyperplasia without toxicity or side effects, and as a health functional food composition. can be used
도 1 본 발명에서 제조된 모새나무열매 추출물의 파우더 및 액상을 나타낸다.Figure 1 shows the powder and liquid phase of the extract of the fruit of Mosaenamu prepared in the present invention.
도 2은 모새나무 추출물에 대한 전립선 상피세포주(LNCaP) 생존율을 나타낸 그래프이다.Figure 2 is a graph showing the survival rate of the prostate epithelial cell line (LNCaP) for the moss extract.
도 3은 테스토스테론에 의한 전립선 상피세포주(LNCaP)에서 모새열매 추출물 처리에 따른 전립선 특이효소(5α-reductase 2), 디히드로테스토스테론(DHT), 및 전립선 특이항원(PSA)의 함량에 미치는 영향을 나타낸 그림이다. Figure 3 shows the effect on the contents of prostate-specific enzyme (5α-reductase 2), dihydrotestosterone (DHT), and prostate-specific antigen (PSA) according to the treatment of the robin fruit extract in the prostate epithelial cell line (LNCaP) by testosterone. It is a picture.
도 4은 테스토스테론에 의한 전립선 상피세포주(LNCaP)에서 모새열매 추출물 처리에 따른 안드로겐 수용체(AR) 단백질 발현에 미치는 그림이다. Figure 4 is a picture showing the androgen receptor (AR) protein expression according to testosterone-induced prostatic epithelial cell line (LNCaP) treatment with moss berry extract.
도 5은 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새열매 추출물 처리에 따른 전립선 조직 비대 억제 효과를 나타낸 그림이다.Figure 5 is a picture showing the effect of inhibiting prostate tissue hypertrophy according to the treatment of the moss berry extract in a testosterone-induced benign prostatic rat model.
도 6은 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새열매 추출물 처리에 따른 전립선 조직에서 5-알파환원효소 함량 억제 효과를 나타낸 그림이다.Figure 6 is a picture showing the effect of inhibiting the content of 5-alpha reductase in prostate tissue according to the treatment of the moss berry extract in a rat model of prostatic hyperplasia induced by testosterone.
도 7은 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새열매 추출물처리에 따른 전립선 조직 단면 염색 사진을 나타내는 그림이다.7 is a diagram showing a cross-sectional staining photograph of prostate tissue according to the treatment of a moss fruit extract in a testosterone-induced benign prostatic rat model.
본 발명은 모새나무 열매 추출물을 유효성분으로 함유하는 전립선 비대증 예방 또는 치료용 조성물에 관한 것으로 구체적인 실시예 및 비교예를 통하여 본 발명의 구성 및 효과를 보다 상세히 설명한다. The present invention relates to a composition for preventing or treating benign prostatic hyperplasia containing an extract of the fruit of moss japonica as an active ingredient, and the configuration and effects of the present invention will be described in more detail through specific examples and comparative examples.
1. 모새나무 열매 추출물의 제조1. Manufacture of mosae tree fruit extract
모새나무 열매 추출물은 수분을 제거하여 건조된 모새나무 열매 5kg에 증류수 50L를 첨가하고, 3시간 동안 100℃로 가열, 추출하였다. 여과하고 감압 및 농축하였다. 농축된 열수추출물은 동결건조기(freeze dryer)를 이용하여 -40℃ ~ 40℃에서 72시간 동안 동결, 건조시켰다. Mosae tree fruit extract was extracted by adding 50 L of distilled water to 5 kg of dried Moss tree fruit by removing moisture, and heating at 100 ° C. for 3 hours. Filtered, reduced pressure and concentrated. The concentrated hot-water extract was frozen and dried at -40 ° C to 40 ° C for 72 hours using a freeze dryer.
이상의 방법으로 모새나무 열매 열수추출물 53.0g(10.6%)를 수득하여 하기 전립선 비대증 예방 또는 치료와 관련된 실험의 시료로 사용하였다. 도 1은 본 발명에서 제조된 모새나무 열매 추출물의 파우더 및 액상을 나타낸다.In the above method, 53.0 g (10.6%) of the hot water extract of the fruit of mosae tree was obtained and used as a sample for the following experiments related to the prevention or treatment of benign prostatic hyperplasia. Figure 1 shows the powder and liquid phase of the extract of the fruit of the moss tree prepared in the present invention.
2. 모새나무 열매 추출물의 세포 독성 확인2. Confirmation of cytotoxicity of the fruit extract of mosae tree
본 발명의 모새나무 열매 추출물의 효과를 확인하기에 앞서, 세포 독성을 나타내지 않으면서 효과를 낼 수 있는 적합한 농도를 결정하기 위해 세포 독성 검사를 수행하였다. 도 2은 모새나무 추출물에 대한 전립선 상피세포주(LNCaP) 생존율을 나타낸 그래프이다.Prior to confirming the effect of the extract of the fruit extract of the present invention, a cytotoxicity test was performed to determine an appropriate concentration capable of producing an effect without exhibiting cytotoxicity. Figure 2 is a graph showing the survival rate of the prostate epithelial cell line (LNCaP) for the moss extract.
모새 열매 추출물의 세포 독성을 확인하기 위하여, 전립선암 세포에 추출물을 처리한 후 세포 생존율을 확인하였다. 전립선암 세포주인 LNCaP 세포(한국세포주은행 21740)를 10% FBS(fetal bovine serum), 1% 페니실린 및 스트렙토마이신이 첨가된 RPMI-1640(Gibco) 배지에서 37℃, 5% CO2 조건하에서 배양하였다. In order to confirm the cytotoxicity of the moss fruit extract, prostate cancer cells were treated with the extract and then the cell viability was confirmed. LNCaP cells (Korea Cell Line Bank 21740), a prostate cancer cell line, were cultured in RPMI-1640 (Gibco) medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin and streptomycin at 37°C and 5% CO 2 conditions. .
상기 배양세포를 96-웰 플레이트에 1x105 cell/well의 농도로 분주하고 배양하고 12시간 후 본 발명에서 제조한 모새나무 열매 추출물을 각각 0, 10, 50, 100㎍/㎖의 농도로 처리하였다. 24시간 배양 후 EZCytox 용액을 각 웰 당 10μL씩 첨가하고 plate reader를 이용하여 450nm에서 흡광도를 측정하여 독성을 확인하였다. 도 2에 나타낸 바와 같이, 모새나무 열매 추출물은 50㎍/㎖까지 세포 독성이 없음을 확인하였다. The cultured cells were dispensed in a 96-well plate at a concentration of 1x10 5 cell/well, cultured, and after 12 hours, the extract of the fruit of the moss tree prepared in the present invention was treated at a concentration of 0, 10, 50, and 100 μg/ml, respectively. . After culturing for 24 hours, 10 μL of EZCytox solution was added to each well, and toxicity was confirmed by measuring absorbance at 450 nm using a plate reader. As shown in FIG. 2, it was confirmed that the extract of the fruit of the moss tree had no cytotoxicity up to 50 μg/ml.
3. 모새나무 열매 추출물의 5α-reductase, DHT 및 PSA 함량 확인3. Confirmation of 5α-reductase, DHT and PSA contents of the fruit extract
도 3은 테스토스테론에 의한 전립선 상피세포주(LNCaP)에서 모새열매 추출물 처리에 따른 전립선 특이효소(5α-reductase 2), 디히드로테스토스테론(DHT), 및 전립선 특이항원(PSA)의 함량에 미치는 영향을 나타낸 그림이다. Figure 3 shows the effect on the contents of prostate-specific enzyme (5α-reductase 2), dihydrotestosterone (DHT), and prostate-specific antigen (PSA) according to the treatment of the robin fruit extract in the prostate epithelial cell line (LNCaP) by testosterone. It is a picture.
전립선 세포에 테스토스테론(Testosterone)과 모새나무 열매 추출물을 처리한 후 5-알파환원효소(5α-reductase), 디하이드로테스토스테론(dihydrotestosterone, DHT) 및 전립선 특이항원(PSA)의 함량을 확인하여 모새나무 열매 추출물이 전립선 비대증 질환에 효과가 있는지 확인하였다. LNCaP 세포를 6 웰 플레이트에 1x106 cells/well의 농도로 분주하고 37℃에서 배양하였다. After treating prostate cells with testosterone and extracts from the fruit of mosae tree, the contents of 5-alpha-reductase, dihydrotestosterone (DHT) and prostate-specific antigen (PSA) were checked. It was confirmed whether the extract was effective in prostatic hyperplasia disease. LNCaP cells were dispensed in a 6-well plate at a concentration of 1x10 6 cells/well and cultured at 37°C.
24시간 후 세포에 테스토스테론 1μM을 처리하고, 동시에 상기 제조한 열수추출물 0, 5, 10, 50㎍/㎖ 농도로 처리한 후 24시간 배양 후 수거하였다. 양성 대조군(BPH)으로는 추출물은 처리하지 않고 테스토스테론 1μM만을 처리하였으며, 대조군으로는 추출물 대신 전립선 비대증 치료제인 Finasteride 10μM을 테스토스테론 1μM과 함께 처리하였다. 무처리군을 음성 대조군(Control)으로 하였다. After 24 hours, the cells were treated with 1 μM of testosterone, and simultaneously treated with the prepared hot-water extract at concentrations of 0, 5, 10, and 50 μg/ml, and then harvested after 24 hours of culture. As a positive control (BPH), only 1 μM of testosterone was treated without extract, and as a control, 10 μM of Finasteride, a treatment for prostatic hyperplasia, was treated with 1 μM of testosterone instead of the extract. The untreated group was used as a negative control group.
배양후 수거한 세포에 phosphate-buffered saline pH7.4(Gibco, USA)를 넣어 Microcentrifuge Sample Pestle로 균질화를 실시하였다. 세포 균질액은 4℃, 5,000 rpm에서 5분간 원심분리(Mikro 200R, Hettich, USA)하여 상등액을 분리하였으며, 각 ELISA KIT(CUSABIO, USA)를 이용하여 5-알파환원효소, 디하이드로테스토스테론 및 전립선 특이항원를 측정하였다.Phosphate-buffered saline pH7.4 (Gibco, USA) was added to the cells harvested after culture, and homogenization was performed with a Microcentrifuge Sample Pestle. The cell homogenate was centrifuged at 4℃ and 5,000 rpm for 5 minutes (Mikro 200R, Hettich, USA) to separate the supernatant, and 5-alpha reductase, dihydrotestosterone and prostate were separated using each ELISA KIT (CUSABIO, USA). Specific antigens were measured.
도 3에 나타낸 바와 같이, 대조군에 비하여 모새 열매 추출물을 50㎍/㎖로 처리한 세포군에서 5-알파환원효소 활성을 14% 억제함을 확인하였다. 아울러, 대조군에 비하여 모새 열매 추출물을 5, 10, 50㎍/㎖로 처리한 세포군에서 디하이드로테스토스테론 활성 억제 효과가 우수함을 확인하였다. 반면 모새 열매 추출물을 10, 50㎍/㎖로 처리한 세포군에서 전립선 특이항원의 함량은 감소하는 경향을 보였으나 통계학적 유의성은 관찰되지 않았다. As shown in FIG. 3, it was confirmed that 5-alpha reductase activity was inhibited by 14% in the cell group treated with 50 μg/ml of the seedling fruit extract compared to the control group. In addition, it was confirmed that the inhibitory effect of dihydrotestosterone activity was excellent in the cell groups treated with 5, 10, and 50 μg/ml of the extract of the seedling fruit compared to the control group. On the other hand, the content of prostate-specific antigen in the cell groups treated with 10 and 50 μg/ml of the moss fruit extract tended to decrease, but no statistical significance was observed.
4. 모새나무 열매 추출물의 안드로겐 수용체(AR) 발현 억제 효과 확인4. Confirmation of the Androgen Receptor (AR) Expression Inhibiting Effect of the Fruit Extract of Mosaic Tree
도 4는 테스토스테론에 의한 전립선 상피세포주(LNCaP)에서 모새열매 추출물 처리에 따른 안드로겐 수용체(AR) 단백질 발현에 미치는 결과를 나타낸 그림이다. 전립선 상피세포(LNCaP)에 테스토스테론(Testosterone)과 모새나무 열매 추출물을 처리한 후 안드로겐 수용체(AR)의 단백질 발현을 확인하여 모새나무 열매 추출물이 전립선 비대증 질환에 효과가 있는지 확인하였다. LNCaP 세포를 6-웰 플레이트에 1x106의 농도로 분주하고 37℃에서 배양하였다. Figure 4 is a picture showing the results of testosterone-induced androgen receptor (AR) protein expression in prostate epithelial cell line (LNCaP) according to the treatment of moss berry extract. Prostate epithelial cells (LNCaP) were treated with testosterone and moss fruit extract, and the protein expression of androgen receptor (AR) was confirmed to confirm whether the moss fruit extract was effective in prostatic hyperplasia. LNCaP cells were dispensed in a 6-well plate at a concentration of 1x10 6 and cultured at 37°C.
24시간 후 세포에 테스토스테론 1μM을 처리하고, 동시에 상기 제조한 열수추출물 0, 5, 10, 50㎍/㎖ 농도로 처리한 후 24시간 배양 후 수거하였다. 양성 대조군(BPH)으로는 추출물은 처리하지 않고 테스토스테론 1μM만을 처리하였으며, 대조군(Fina)으로는 추출물 대신 전립선 비대증 치료제인 Finasteride 10μM을 테스토스테론 1μM과 함께 처리하였다. 무처리군을 음성 대조군(Control)으로 하였다. After 24 hours, the cells were treated with 1 μM of testosterone, and simultaneously treated with the prepared hot-water extract at concentrations of 0, 5, 10, and 50 μg/ml, and then harvested after 24 hours of culture. As a positive control group (BPH), only 1 μM of testosterone was treated without the extract, and as a control group (Fina), 10 μM of Finasteride, a treatment for prostatic hyperplasia, was treated with 1 μM of testosterone instead of the extract. The untreated group was used as a negative control group.
배양 후 수거한 세포에 RIPA buffer(50 mM Tris-HCl, pH 8.0, with 150 mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate, with a protease inhibitor cocktail)를 처리하여 단백질을 분리하였다. Cells harvested after culture are treated with RIPA buffer (50 mM Tris-HCl, pH 8.0, with 150 mM sodium chloride, 1% NP-40, 0.5% sodium deoxycholate, and 0.1% sodium dodecyl sulfate, with a protease inhibitor cocktail) to isolate the protein.
분리된 단백질은 단백질 분석 키트(Bio-Rad protein assay kit, Bio-Rad, USA)를 이용하여 정량후 정량한 단백질을 10% 폴리아크릴아마이드 겔에서 SDS-PAGE를 수행하고 PVDF 멤브레인으로 단백질을 전사하였다. The separated protein was quantified using a protein assay kit (Bio-Rad protein assay kit, Bio-Rad, USA), and the quantified protein was subjected to SDS-PAGE on a 10% polyacrylamide gel, and the protein was transferred to a PVDF membrane. .
이후 PVDF 멤브레인을 5% 스킴밀크(skim milk)에 1시간 정도 반응시켜 표면의 비특이적 결합 단백질들을 제거하고, 1차 항체인 항-AR 항체 또는 항-β-actin 항체를 이용하여 4℃에서 하루 동안 반응시켰다. 이후 2차 항체를 1시간 동안 상온에서 처리하고 ECL kit(Thermo scientific, 미국)를 이용하여 AR의 단백질 발현 정도를 확인하였다. 도 4에 나타낸 바와 같이, 대조군에 비하여 모새 열매 추출물을 5, 10, 50㎍/㎖을 처리한 세포군에서 AR 단백질 발현 억제 효과가 현저히 우수함을 확인하였다. Thereafter, the PVDF membrane is reacted with 5% skim milk for about 1 hour to remove non-specific binding proteins on the surface, and the primary antibody, anti-AR antibody or anti-β-actin antibody, is used for one day at 4 ° C. reacted Thereafter, the secondary antibody was treated at room temperature for 1 hour, and the level of AR protein expression was confirmed using an ECL kit (Thermo scientific, USA). As shown in FIG. 4, it was confirmed that the AR protein expression inhibitory effect was remarkably superior to the control group in the cell group treated with 5, 10, or 50 μg/ml of the moss fruit extract.
5. 전립선 비대증 동물 제작 및 전립선 무게 측정5. Production of animals with benign prostatic hyperplasia and measurement of prostate weight
도 5는 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새나무 열매 열수추출물 처리에 따른 전립선 조직 비대 억제 효과를 나타낸 그림이다. 9주령(몸무게 330g 이하)의 수컷 SD 쥐를 1주일간 순화 사육한 후, 테스토스테론 프로피오네이트(testosterone propionate, TP) 3mg/kg 용량을 옥수수 오일에 녹여 피하에 4주간 주사하여 전립선 비대증을 유도하였다. Figure 5 is a picture showing the effect of inhibiting prostate tissue hypertrophy according to treatment with hot water extract of moss tree fruit in a rat model of prostatic hyperplasia induced by testosterone. Male SD rats aged 9 weeks (weighing less than 330 g) were domesticated for 1 week, and then 3 mg/kg of testosterone propionate (TP) was dissolved in corn oil and subcutaneously injected for 4 weeks to induce prostate hyperplasia.
모새나무 열매 열수 추출물을 50, 100mg/kg으로 4주간 경구 투여하였으며, 양성대조군으로 전립선 비대증 치료에 사용되고 있는 5α-환원효소 억제제인 피나스테리드(Finasteride, FI)를 5mg/kg 용량으로 경구 투여하였다. 실험 시작일로부터 일주일 간격으로 총 5회 몸무게를 측정하고, 마지막 투약 및 처리가 끝난 다음날 랫드를 희생시킨 후, 각 군(정상군; Sham, 전립선 비대증 유발군(음성대조군); BPH, 전립선 비대증 유발군+피나스테리드 투여군(양성대조군); FI, 전립선 비대증 유발군+모새나무 열매 추출물50mg/kg 투여군; VBF50, 전립선 비대증 유발군+모새나무 열매 추출물 100mg/kg 투여군; VBF100으로부터 전립선을 적출하여 그 무게를 측정하고 좌측 전립선조직을 적출하여 조직에서 5α-환원효소를 측정하였다. 전립선증식억제효과(%)는 다음 식에 따라 계산된다. The hot-water extract of the fruit of mosae tree was orally administered at 50 and 100 mg/kg for 4 weeks, and finasteride (FI), a 5α-reductase inhibitor used for the treatment of benign prostatic hyperplasia, was orally administered at a dose of 5 mg/kg as a positive control group. After measuring the body weight a total of 5 times at intervals of one week from the start of the experiment, and sacrificing the rats the day after the last administration and treatment, each group (normal group; Sham, prostatic hyperplasia induced group (negative control group); BPH, prostatic hyperplasia induced group + finasteride administration group (positive control group); FI, benign prostatic hyperplasia group + moss fruit extract 50mg/kg administration group; VBF50, benign prostatic hyperplasia induction group + mosae tree fruit extract 100mg/kg administration group; prostates were removed from VBF100 and weighed The left prostate tissue was removed and 5α-reductase was measured in the tissue, and the prostate growth inhibitory effect (%) was calculated according to the following equation.
전립선증식억제효과(%) = 100 - {(모새 열매 투여군 또는 양성대조군 전립선무게 - 정상군 전립선무게)*100 / (음성대조군 전립선무게 - 정상군 전립선무게)}Prostatic hyperplasia inhibitory effect (%) = 100 - {(prostate weight in the seed-treated group or positive control group - normal prostate weight) * 100 / (negative control prostate weight - normal prostate weight)}
도 5에 나타낸 바와 같이, TP로 전립선 비대증을 유발한 군(BPH) 보다 모새열매 추출물을 투여군(VBF50 및 VBF100)에서 전립선의 크기가 감소하는 것을 확인하였다. As shown in FIG. 5, it was confirmed that the size of the prostate was reduced in the groups (VBF50 and VBF100) administered with the mulberry fruit extract than in the group (BPH) in which prostatic hyperplasia was induced by TP.
표 1은 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새열매 추출물 처리에 따른 전립선 조직 무게 표를 나타낸다. 표 1에서 나타낸 바와 같이, 전립선 비대증 유도 동물모델에 모새나무 열매 추출물을 경구 투여하여 전립선의 항비대 효과를 측정한 결과, 전립선 크기는 대조군에서 0.30±0.01로 정상군의 0.18±0.02에 비해 29.7%가 증가하였다. 전립선 비대 증식억제효과는 양성대조군에서 73.56%를 나타냈으며, 모새나무 열매 50 또는 100mg/kg 투여군에서는 각각 20.84%, 28.75%로 농도 의존적으로 억제효과를 보였다.Table 1 shows a table of prostate tissue weights according to the treatment of moss berry extract in a testosterone-induced benign prostatic rat model. As shown in Table 1, as a result of measuring the anti-hypertrophy effect of the prostate by orally administering the fruit extract of mosae tree to an animal model inducing benign prostatic hypertrophy, the prostate size was 0.30 ± 0.01 in the control group, 29.7% compared to 0.18 ± 0.02 in the normal group. has increased. Prostate hyperplasia growth inhibitory effect was 73.56% in the positive control group, and 20.84% and 28.75% respectively in the group administered with 50 or 100mg/kg of moss fruit, showing inhibitory effects in a concentration-dependent manner.
groupgroup | 전립선무게(g/100BW)Prostate weight (g/100BW) | 증식억제효과(%)Growth inhibitory effect (%) |
ShamSham | 0.18±0.02 *** 0.18±0.02*** | -- |
BPHBPH | 0.30±0.01 0.30±0.01 | -- |
FIFI | 0.21±0.02 *** 0.21±0.02*** | 73.5673.56 |
VBF50VBF50 | 0.27±0.02 0.27±0.02 | 20.8420.84 |
VBF100VBF100 | 0.26±0.01 * 0.26±0.01* | 28.7528.75 |
6. 알파환원효소 측정도 6은 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새열매 추출물 처리에 따른 전립선 조직에서 5-알파환원효소 함량 억제 효과를 나타낸 그림이다. 희생된 랫의 전립선 조직은 생리식염수로 1회 세척하고 4배의 phosphate-buffered saline pH7.4(Gibco, USA)를 넣어 Wise Stir homogenizer(Daihan Scientific, Korea)로 1분간 균질화를 실시하였다. 전립선 조직 균질액은 4℃, 5,000 rpm에서 5분간 원심분리(Mikro 200R, Hettich, USA)하여 상등액을 분리하였으며, 5-알파환원효소 ELISA KIT(CUSABIO, USA)를 이용하여 5-알파환원효소를 측정하였다.6. Alpha-reductase measurement Figure 6 is a diagram showing the inhibitory effect of 5-alpha-reductase content in prostate tissue according to the treatment of the moss fruit extract in a rat model of prostatic hyperplasia induced by testosterone. The prostate tissue of the sacrificed rat was washed once with physiological saline and homogenized for 1 minute with a Wise Stir homogenizer (Daihan Scientific, Korea) after adding 4 times phosphate-buffered saline pH7.4 (Gibco, USA). Prostate tissue homogenate was centrifuged at 4°C and 5,000 rpm for 5 minutes (Mikro 200R, Hettich, USA) to separate the supernatant. measured.
도 6에 나타낸 바와 같이, 정상군, 음성대조군, 양성대조군, 모새나무열매 50 및 100mg/kg에서 각각 62.79±17.36, 617.45±82.86, 232.8±52.48, 446.0±66.76, 314.9±67.47로 측정되었다. 양성대조군과 모새나무 열매 100mg/kg 투여군에서 유의적인 감소를 나타내었다.As shown in FIG. 6, 62.79 ± 17.36, 617.45 ± 82.86, 232.8 ± 52.48, 446.0 ± 66.76, and 314.9 ± 67.47 in the normal group, negative control group, positive control group, and 50 and 100 mg / kg of mosae tree fruit, respectively. A significant decrease was shown in the positive control group and the group administered with 100 mg/kg of moss fruit.
6. 전립선 조직병리학적 관찰6. Prostate histopathological observation
도 7은 테스토스테론으로 유도한 전립선 비대증 랫드 모델에서 모새열매 추출물처리에 따른 전립선 조직 단면 염색 사진을 나타내는 그림이다. 희생된 랫의 우측 전립선 조직은 10% 중성 포르말린에서 고정한 후 파라핀 포매하여 절편을 제작하여, Hematoxylin-Eosin 염색을 시행한 후, 광학 현미경으로 전립선의 상피세포와 기질의 조직학적 변화를 관찰하였다.7 is a diagram showing a cross-sectional staining photograph of prostate tissue according to the treatment of a moss fruit extract in a testosterone-induced benign prostatic rat model. The right prostate tissues of the sacrificed rats were fixed in 10% neutral formalin, paraffin-embedded and sectioned, and stained with Hematoxylin-Eosin. Histological changes in the epithelial cells and stroma of the prostate were observed under an optical microscope.
도 7에 나타낸 바와 같이, 전립선 비대증을 유도한 결과 정상군에 비해 음성대조군(BPH)에서 전립선상피조직이 두꺼워지고 다수의 폴립이 관찰되었으나 모새나무 열매 50 또는 100mg/kg 투여군에서는 상피조직이 얇아지고 폴립도 적어진 것을 관찰하였다. As shown in FIG. 7, as a result of inducing prostatic hyperplasia, the prostate epithelial tissue became thicker and a number of polyps were observed in the negative control group (BPH) compared to the normal group, but in the group administered with 50 or 100 mg / kg of moss fruit, the epithelial tissue became thin and A decrease in polyps was also observed.
8. 모새나무 열매 추출물을 이용한 약제 또는 건강식품 제조8. Manufacture of pharmaceuticals or health foods using extracts from the fruit of mosae tree
모새나무 열매 추출물을 유효성분으로 하는 전립선 비대증 예방 또는 치료용 약학제제 또는 건강식품은 정제 및 캡슐제, 연질 캡슐제, 과립제, 액제 형태로 제조될 수 있다. 또 다른 적절한 실시 형태에 따르면, 상기 약학제제 또는 건강식품은 음료 첨가제로 제조할 수 있다. Pharmaceutical preparations or health foods for the prevention or treatment of prostatic hyperplasia using the fruit extract of moss tree as an active ingredient may be prepared in the form of tablets and capsules, soft capsules, granules, and liquids. According to another suitable embodiment, the pharmaceutical formulation or health food may be prepared as a beverage additive.
상기 전립선 비대증 예방 또는 치료용 약학제제 또는 건강식품은 상기 모새나무 열매 추출물을 유효성분으로 포함하는 조성물이 0.01 내지 99.9중량%로 포함되도록 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸, 경피제, 좌제 또는 멸균 주사용 액으로 제형화하여 제조할 수 있다.The pharmaceutical formulation or health food for preventing or treating prostatic hyperplasia is a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol such that the composition containing the mosae tree fruit extract as an active ingredient is contained in an amount of 0.01 to 99.9% by weight , It can be prepared by formulating into a transdermal preparation, a suppository or a sterile injectable solution.
멸균주사액의 경우, 상기 약학 조성물이 0.01 내지 99.9중량%로 포함하도록 하고 정제수 또는 포도당을 99.9 내지 0.01 중량%를 혼합하여 제조 가능하며, 캡슐제의 경우, 상기 약학조성물을 동결 건조하여 0.01 내지 99.9중량%로 포함하도록 하고, 비타민제, 칼슘제을 99.9 내지 0.01 중량%를 혼합하여 제조 가능하다.In the case of a sterile injection solution, the pharmaceutical composition can be prepared by mixing 99.9 to 0.01% by weight of purified water or glucose with 0.01 to 99.9% by weight of the pharmaceutical composition, and in the case of capsules, freeze-drying the pharmaceutical composition to 0.01 to 99.9% by weight %, and can be prepared by mixing 99.9 to 0.01% by weight of vitamins and calcium.
상기 제조된 약학조성물의 1일당 투여량은 상기 추출물이 10 내지 2000mg/kg 체중 포함되는 함량으로 제공하며, 또한 상기 약학 조성물을 0.01 내지 99.9중량%로 포함하는 전립선 비대증 예방 또는 치료용 약학제제 또는 건강식품으로 제조가능하다.The daily dosage of the pharmaceutical composition prepared above is provided in an amount of 10 to 2000 mg/kg body weight of the extract, and a pharmaceutical preparation for preventing or treating enlargement of the prostate or health containing 0.01 to 99.9% by weight of the pharmaceutical composition. can be made into food.
본 발명은 모새 열매 추출물을 유효성분으로 하는 전립선 비대증 예방 또는 억제용 조성물로써 테스토스테론으로 유도한 전립선 세포(LNCaP) 및 전립선 비대증 랫드 모델에서 우수한 전립선 비대증 예방 또는 억제 효과에 도움이 되므로 산업상 이용가능성 있다.The present invention is a composition for preventing or inhibiting prostatic hyperplasia, which contains an extract of moss fruit as an active ingredient, and is useful for preventing or inhibiting prostatic hyperplasia in testosterone-induced prostate cells (LNCaP) and an enlarged prostate rat model, so it has industrial applicability. .
Claims (5)
- 모새나무(Vaccinium Bracteatum Thunb.) 열매 열수추출물을 유효성분으로 포함하는 것을 특징으로 하는 전립선 비대증 예방 또는 치료용 약학 조성물A pharmaceutical composition for preventing or treating prostatic hyperplasia, comprising a hot water extract of the fruit of Vaccinium Bracteatum Thunb.
- 청구항 1에 있어서, 상기 조성물을 0.01 내지 99.9중량% 함유하는 것을 특징으로 하는 전립선 비대증 예방 또는 치료용 약학 조성물The pharmaceutical composition for preventing or treating prostatic hyperplasia according to claim 1, which comprises 0.01 to 99.9% by weight of the composition.
- 청구항 1에 있어서, 상기 조성물은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 경피제, 좌제 또는 멸균 주사용 액 중에서 선택되는 어느 하나의 형태로 제형화된 것을 특징으로 하는 전립선 비대증 예방 또는 치료용 약학 조성물The method according to claim 1, wherein the composition is formulated in any one form selected from powders, granules, tablets, capsules, suspensions, emulsions, syrups, transdermal preparations, suppositories, or sterile injectable liquids. Prevention of prostatic hyperplasia, characterized in that or a pharmaceutical composition for treatment
- 모새나무(Vaccinium Bracteatum Thunb.) 열매 열수추출물을 유효성분으로 포함하는 것을 특징으로 하는 전립선 비대증 예방 또는 개선용 기능성 식품조성물Functional food composition for preventing or improving prostatic hyperplasia, characterized in that it contains a hot water extract of the fruit of mosae tree ( Vaccinium Bracteatum Thunb.) as an active ingredient
- 청구항 4에 있어서, 기능성 식품조성물은 정제 및 캡슐제, 연질 캡슐제, 과립제, 액제 중에서 선택되는 어느 하나의 형태로 제조되는 것을 특징으로 하는 전립선 비대증 예방 또는 개선용 기능성 식품조성물The functional food composition for preventing or improving prostatic hyperplasia according to claim 4, wherein the functional food composition is prepared in any one form selected from tablets and capsules, soft capsules, granules, and liquids.
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KR1020210157710A KR102388984B1 (en) | 2021-11-16 | 2021-11-16 | PHARMACEUTICAL COMPOSITION CONTAINING VACCINIUM BRACTEATUM Thunb. EXTRACT FOR Treating and Preventing Benign Prostate Hyperplasia |
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KR101230644B1 (en) | 2012-02-16 | 2013-02-06 | 주식회사 더마프로 | Composition useful for human skin whitening comprising vaccinium bracteatum thunb extract |
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KR101829583B1 (en) | 2017-06-01 | 2018-02-20 | 재단법인 전남생물산업진흥원 | Pharmaceutical composition having anti-depressant activity and preparation method using Vaccinium bracteatum Thunb. of the same. |
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