KR101863731B1 - Composition for preventing, improving or treating prostate disease comprising extract of Cardamine scutata as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating, or treating prostate disease containing a Cardamine scutata Thunb. extract as an active ingredient. The Cardamine scutata Thunb. extract as the active ingredient decreases the weight of the enlarged prostate due to the prostatic hypertrophy (BPH), suppresses expression of androgenic receptors or prostate specific antigen proteins, and significantly reduces the levels of testosterone and dihydrotestosterone (DHT) in serum and prostate.

Description

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a composition for preventing, improving or treating a prostate disease,

The present invention relates to a composition for preventing, ameliorating or treating a prostate disease, which comprises a large bolecups extract as an active ingredient.

Prostate is the organ that produces various secretions to keep the urethra from drying out. It is closely related to the male hormone, androgens. As the secretion increases, the size of the prostate increases in the 20s and 30s. This is called benign prostatic hyperplasia and it is difficult to cure it because the risk of side effects is high even if the size is returned to the normal level by the hormone treatment, and soon after stopping the hormone treatment, it returns to the original state.

Representative diseases that occur in the prostate gland include enlarged prostate, chronic prostatitis, and prostate cancer. Prostate cancer is a disease caused by bacterial infections, inflammatory or chronic pain, and prostate cancer is a malignant tumor in the relatively outer part of the prostate gland. According to a US study, It accounts for about 35% of all male cancer patients and is said to increase by 2-3% every year. The incidence in Korea is three per 100,000 people, yet it is not worrisome yet, but it is gradually increasing with the western dietary life and the rapidly aging population.

On the other hand, benign prostatic hyperplasia (BPH) is a disease in which the urethral prostate "ganglion" is enlarged due to aging and the imbalance of sex hormones, resulting in urinary symptoms due to the pressure of the urethra. It is a male disease. Recently, the rate of growth of patients with hyperplasia of the prostate in Korea has been steadily increasing to more than 20% every year. It is caused by excessive proliferation of smooth muscle and epithelial part of the prostate transplantation part, , Which is known to be due to anatomical occlusion by the enlarged prostate and functional occlusion by the? 1 -receptor which contracts the prostate smooth muscle. The main symptoms of occlusion include urinary tract weakness or weak urine stem or urine, delayed urination to wait for a long time to start urinating, discontinuation of urination to stop irrespective of urinary incontinence, terminal urination dropping like urine drops, This is representative. It is known that the testosterone (T) and the dihydrotestosterone (DHT) are involved in the typical mechanism of the various mechanisms of prostate hypertrophy.

On the other hand, Cardamine scutata is a plant native to the south of Gangwon Province and grows in streams or wetlands. Leaves are alternate phyllotaxis, 3 ~ 9 small lobes and odd phyllotaxic compound leaves, similar to almost storks, but with little lobes and round, especially large lobules with a width of 2.5cm. Leaflets have dull end, irregularly split edges, no hairs on both sides, or some. Flower is bloomed in May ~ June, and twenty white cross-shaped corolla is formed at the tip of the branch and main stem. The calyx is long elliptical, 2mm long, petal is wide spatulate shape, and length is 4mm. One pistil grows to be 2 ~ 3cm long. When ripened, it rips into two pieces and it dries and the seeds protrude and have no hair. Its height is 20cm. The main stem is soft and almost hairless. It grows diagonally at the upper part divided into several.

TECHNICAL FIELD The present invention relates to a composition for preventing, ameliorating, or treating a prostate disease using natural extracts. Examples of techniques related to a testosterone 5α-reductase inhibitor and an androgen receptor binding inhibitor, an antihistamines agent, a wool cosmetic agent and a prostate hypertrophy inhibitor are disclosed in Japanese Patent No. 5191525 And Korean Patent No. 1653492 discloses a composition for treating inflammation or enlargement of the prostate gland comprising an extract of beech leaf blossom or a compound isolated therefrom as an active ingredient, A composition for preventing, ameliorating, or treating a prostate disease, which comprises the composition as described above, has not been disclosed.

The present invention provides a composition for preventing, ameliorating or treating a prostate disease, which comprises an extract of a large boleciaceae as an active ingredient. The present invention provides a composition for preventing, improving or treating a prostate disease, (BPH) reduces the weight of the enlarged prostate, inhibits the expression of androgenic receptor or prostate specific antigen protein, and inhibits the expression of serum and prostate-specific testosterone and DHT The content can be remarkably reduced, thereby completing the present invention.

In order to achieve the above object, the present invention is large Stork cob (Cardamine scutata extract as an active ingredient. The present invention also provides a pharmaceutical composition for preventing or treating prostate disease.

The present invention also provides an agent for improving dysuria caused by hypertrophy of the prostate, which comprises the above pharmaceutical composition as an active ingredient.

The present invention also relates to a method for the treatment of cardamom scutata extract as an active ingredient. The present invention also provides a health functional food composition for preventing or ameliorating a prostate disease.

The present invention relates to a composition for preventing, ameliorating or treating a prostate disease, which comprises a large boleciaceae extract as an active ingredient, wherein the large borealis extract of the present invention is an active ingredient of a prostate Progesterone or prostate specific antigen protein because it significantly reduces the amount of testosterone and DHT in the serum and the prostate as well as inhibits the expression of the androgenic receptor or the prostate specific antigen protein, Can be effectively applied to prostate diseases of prostate cancer.

FIG. 1 shows the results of confirming the cell survival rate (%) according to the treatment of the extract of the great thistle bark of the present invention. NC is a negative control with no treatment.
2 is a Western blotting result showing the effect of inhibiting the expression of an androgenic receptor (AR) and a prostate specific antigen (PSA) protein of a large Staphylococcus aureus extract. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 3 is a graph showing the results of Western blotting of FIG. 2, wherein A represents the expression level of an androgenic receptor (AR) and B represents the expression level of a prostate specific antigen (PSA) protein will be. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 4 is a graph showing the weight change of the prostate according to the administration of the extract of the great white stork of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 5 is a graph showing the change in DHT content in serum according to the administration of the extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 6 shows the results of confirming the change in the content of testosterone in serum according to the administration of the extract of the great thistle of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 7 shows H & E staining results of the prostate tissue according to the administration of the large stallion cold extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 8 shows the results of the ALT and AST contents changes in the liver according to the administration of the extract of the great white stork from the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + CS was associated with prostatic hyperplasia and the treatment of large Staphylococcus aureus extract; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.

The present invention relates to a method for the treatment of Cardamine The present invention relates to a pharmaceutical composition for preventing or treating a prostate disease.

The prostate disease is preferably benign prostatic hyperplasia, prostate cancer or prostatitis, but it is not limited thereto, more preferably it is benign prostatic hyperplasia, and may include hair loss or urination disorder due to enlarged prostate.

The above-mentioned large balsam extract can be produced by a method including, but not limited to, the following steps:

(1) extracting a large stallion with an extraction solvent;

(2) filtering the extract of step (1); And

(3) The step of extracting the filtered extract of step (2) by concentration under reduced pressure and drying.

In the step (1), the extraction solvent is preferably selected from water, a C ₁ -C ₄ lower alcohol or a mixture thereof, more preferably a C ₁ -C ₄ lower alcohol, even more preferably 99% (v / v) methanol, but is not limited thereto.

In the above production method, any conventional method known in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction may be used. It is preferable that the extraction solvent is added by 1 to 20 times the weight of the dried large white stallion, more preferably 5 to 15 times. The extraction temperature is preferably 4 to 50 DEG C, but is not limited thereto. The extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but not always limited thereto. In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in the step (3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

The large Staphylococcus aureus extract can inhibit expression of an androgenic receptor or a prostate specific antigen protein and decrease the content of DHT (dihydrotestosterone) or testosterone in the serum.

The pharmaceutical composition of the present invention may further comprise a carrier, an excipient or a diluent in addition to the above-described large balsam extract.

The pharmaceutical composition of the present invention may be administered orally or parenterally, and it is preferable to use an intravenous or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebral injection method during parenteral administration.

The pharmaceutical composition of the present invention can be prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient. Solid formulations for oral administration include tablets, pills, powders, granules, capsules, and the like, which may contain one or more excipients such as starch, calcium carbonate, sucrose or lactose lactose, gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. Examples of suppositories include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.

The composition according to the present invention is administered in a pharmaceutically effective amount. In the present invention, "pharmaceutically effective amount" means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the composition of the present invention may be varied depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease.

The present invention also relates to an agent for improving dysuria caused by hypertrophy of the prostate, which comprises the above pharmaceutical composition as an active ingredient.

The present invention also relates to a method for the treatment of cardamom scutata extract as an active ingredient. The present invention relates to a health functional food composition for preventing or ameliorating a prostate disease.

The health functional food composition for preventing or ameliorating a prostate disease, which comprises the above-described large anthelmintic extract of the present invention as an active ingredient, may be prepared by any one selected from beverage, ring, tablet, capsule and acid, , And can be suitably prepared according to a conventional method.

Examples of the food to which the present invention can add a large stork extract include dairy products including meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, Beverage, tea, drink, alcoholic beverage, and vitamin complex, and includes all the health functional foods in the conventional sense.

The health functional foods include various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavors, colorants and enhancers (cheese, chocolate etc.), pectic acid and its salts, alkynic acid and its salts, , pH adjusting agents, stabilizers, preservatives, glycerin, alcohols, carbonating agents used in carbonated drinks, and the like. It may also contain flesh for the production of natural fruit juices and vegetable drinks. These components may be used independently or in combination.

The health functional food composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient. The natural carbohydrates are sugar saccharides such as monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, polysaccharides such as dextrin and cyclodextrin, xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  One. Big stork ( outpost , 037-086) Preparation of extract

A large extract of 21.88 mg of Staphylococcus aureus was obtained from Korea Plant Extract Bank at an extraction yield of 14.1% using 99.9% (v / v) methanol grade (HPLC grade) Mu] l of DMSO to give a concentration of 100 mg / ml and used for in vitro experiments.

In addition, a large amount of 7.36 g of Staphylococcus aureus extract, which was extracted from Korea Plant Extract Bank at a yield of 19.20% using 99.9% (v / v) methanol (HPLC grade) , And in vivo .

Example  2. Cells from the treatment of the large Staphylococcus aureus extract Of cell viability  Confirm

In order to examine the effect of the large Horseradish peroxidase extract on LN cap cells, 1 × 10 ⁴ LN cap cells were plated on a 96-well plate and cultured at 37 ° C. for 24 hours. Then, , 50, 100, 200 占 퐂 / ml), and further cultured for 72 hours.

After that, 10 당 of EZ Cytox Assay Kit (DOGEN, Korea) was added to each well, and reacted for about 2 hours. Absorbance was measured at 450 nm with an enzyme-linked immunosorbent assay (ELISA) reader.

As a result, it was confirmed that even when treated with a large extract of Stork bacillus as shown in Fig. 1, cell viability was not affected and thus there was almost no cytotoxicity.

Example  3. Androgenic receptor (AR) and prostate-specific antigen prostate specific antigen ; PSA ) Confirmation of Protein Expression Inhibitory Effect

5 × 10 ⁵ cells were plated on a 6-well plate and cultured for 24 hours. All wells except the control were treated with 100 nM testosterone, And 200 [mu] g / ml), and then cultured for 72 hours at 37 DEG C and 5% CO ₂ . As a positive control, 10 μM Finasteride was used.

After the reaction was completed, the cells were washed twice with PBS, and a protein extraction solvent (20 mM Tris-HCl, 150 mM NaCl, 1 mM Na ₂ EDTA, 1 mM EGTA, 1% NP- 40, 1% sodium deoxycholate, , 2.5 mM sodium pyrophosphate, 1 mM b-glycerophosphate, 1 mM Na ₃ VO ₄ , 1 μg / ml leupeptin (Cell signaling, USA) After centrifugation at 12,000 rpm for 20 minutes at 4 ° C, the supernatant was collected and the protein was quantified with a protein analysis kit (Bio-Rad protein assay kit, Bio-Rad, USA) And 10 μg of the androgen receptor protein and 30 μg of the prostate-specific antigen protein.

As a result, the degree of protein expression was confirmed by Western blotting (FIG. 2), and the amount of expression was quantified and shown in a graph. In the case of the androgen receptor protein (AR), the expression of AR decreased from 50 ㎍ / ㎖ of the extract of the great borealis extract, especially when the concentration of 200 ㎍ / ㎖ was treated with the positive control, Finasteride (Fig. 3A).

As shown in FIG. 3B, the expression of prostate-specific antigen (PSA) protein was decreased when a large-billed cold extract was treated at a concentration of 200 μg / ml.

Example  4. Production and statistical analysis of animal models

After 10-week-old male Wister rats (central laboratory animals) were purified for 1 week, the prostate hypertrophy group (BPH) was injected with 3 mg / kg of testosterone propionate (TP) Respectively. In the group treated with the large anthelmintic extract, 100 mg / kg of a large-sized brucid extract was orally administered before the injection of TP for 4 weeks, and the positive control group was treated with 10 mg / kg of Finasteride, a 5α-reductase inhibitor used as a treatment for benign prostatic hyperplasia, kg < / RTI >

Statistical analysis was performed with ANOVA, with # being statistically significant when compared to normal ( P <0.05); * Indicates a statistically significant difference compared to the BPH group, and P <0.05.

Example  5. Prostate Weighing

The rats were injected subcutaneously with testosterone propionate (TP) for 4 weeks to induce hypertrophy of the prostate gland and to be administered orally together with finasteride (Finasteride), which is used as a treatment for the treatment of large stallions and prostate hyperplasia. After the administration of the drug, sacrificed rat prostate glands were sampled and weighed.

As a result, as shown in FIG. 4, in each experimental group (normal group: NC, prostate hyperplasia-induced group: BPH, prostate hyperplasia group + large bolecrodia extract-treated group: BPH + CS, prostate hypertrophy + The weight of prostate measured by BPH + F group was higher than that of normal group by TP, and the weight of prostate was significantly increased Was significantly decreased.

Example  6. Prostate DHT  change

After the end of the experiment, the separated blood was centrifuged at 4,000 rpm for 20 minutes to check the content of DHT (dihydrotestosterone) in an enlarged prostate animal model. The supernatant was taken from the serum, and the serum was measured using an ELISA kit (ALPCO, Salem) The content of DHT (dihydrotestosterone) was measured.

As a result, as shown in FIG. 5, it was confirmed that the content of DHT (dihydrotestosterone) in serum was statistically significantly decreased in the group administered with the large-billed cold extract.

Example  7. Serum Testosterone  Analysis of content change

After the end of the experiment, the separated blood was centrifuged at 4,000 rpm for 20 minutes, and the supernatant was taken. The serum was analyzed by ELISA kit (Cayman, USA) for testosterone (testosterone ) Were measured.

As a result, as shown in FIG. 6, the testosterone content in serum decreased statistically significantly in the group administered with the large extract of Stork.

Example  8. Histopathological changes of the prostate gland

After the end of the experiment, the extracted prostate gland was fixed with 10% neutral buffered formalin for 24 hours and paraffin embedding was performed. The embryoid tissue was cut to a thickness of 4 μm and cut into sections and stained with H & E (hematoxylin, Sigma-Aldrich, USA) and eosin Y (Sigma-Aldrich) And examined with an optical microscope.

As a result, as shown in FIG. 7, the height and number of epithelial cells were overproduced in the prostatic hyperplasia-induced group, but it was confirmed that the proliferation of the epithelial cells was inhibited in the group treated with the great-anthelmintic extract.

Example  9. Serum ALT and AST  Measure

In order to confirm the toxicity of the large bush extract, the isolated blood was centrifuged at 4,000 rpm for 20 minutes, and then the supernatant was taken to determine the contents of ALT and AST in the serum.

As a result, as shown in FIG. 8, statistically significant changes were observed in the contents of ALT and AST used as general liver toxicity (damage) index in both the group of the patients with the benign prostatic hyperplasia and those of the group of the patients treated with the great anthelmintic extract It was not.

Claims (8)

A pharmaceutical composition for preventing or treating enlargement of the prostate gland comprising Cardain scutata extract as an active ingredient. delete The method of claim 1, wherein the extraction solvent of the large Stork horseradish extract is water, C ₁ ~ C ₄ lower alcohol, or the prevention or treatment of a pharmaceutical composition for prostatic hyperplasia, characterized in that a mixture thereof. The pharmaceutical composition according to claim 1, wherein the large anthelmintic extract inhibits the expression of an androgenic receptor or a prostate specific antigen protein. The pharmaceutical composition for preventing or treating enlargement of the prostate gland according to claim 1, wherein the large anthelmintic extract lowers the content of dihydrotestosterone (DHT) or testosterone in serum. A method for improving urination disorder caused by hypertrophy of the prostate, which comprises the composition of any one of claims 1 to 5 as an active ingredient. A health functional food composition for preventing or ameliorating hyperplasia of the prostate, which comprises an extract of Cardamine scutata as an active ingredient. 8. The method according to claim 7, wherein the Cardamine scutata extract is prepared from any one of a beverage, a ring, a tablet, a capsule, and a powder. &Lt; / RTI &gt;

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