KR101923170B1 - Composition for preventing, improving or treating prostate disease comprising extract of Eriochloa villosa as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating or treating a prostate disease, comprising a mildew extract as an active ingredient. The effective ingredient of the present invention is the extract of N. parapertussis which reduces the weight of the enlarged prostate by BPH and inhibits the expression of androgenic receptor or prostate specific antigen protein , The serum and the amount of testosterone and DHT in the prostate can be significantly reduced.

Description

[0001] The present invention relates to a composition for preventing, improving or treating a prostate disease, which contains an extract of a mulberry as an active ingredient,

The present invention relates to a composition for preventing, ameliorating or treating a prostate disease which comprises a mildew 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 therapy and soon after stopping the hormone treatment.

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, I do cigarettes (Eriochloa villosa ) is a perennial plant, with smooth hairs growing throughout the plant and making bundles. Leaves grow from beak, are slightly dark green, and have long hair with inflorescence. Short hairs also have hairs, and when dried they turn slightly black. It grows in one direction from July to August, and the whole white hair is densely and there is a round base on the base of small flower ears.

Korean Patent No. 1653492 discloses a composition for treating inflammation or enlargement of the prostate gland, which comprises a bee flower leaf extract or a fraction thereof as an active ingredient and is known as a composition for prevention, improvement or treatment of prostate disease, Patent No. 5191525 discloses that at least one natural product selected from olam, Huang umbrella, and Tengshi lotus is known as a testosterone 5α-reductase inhibitor and an androgen receptor binding inhibitor. However, in the present invention, prevention of prostate disease , Compositions for improvement or treatment have not been disclosed.

The present invention provides a composition for preventing, ameliorating or treating a prostate disease comprising an extract of Aspergillus oryzae 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 and inhibits the expression of androgenic receptor or prostate specific antigen protein as well as the content of testosterone and DHT in serum and prostate The present invention has been completed.

In order to achieve the above object, the present invention I cigarettes (Eriochloa villosa 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.

In addition, the present invention is also directed to a method of treating < RTI ID = villosa extract as an active ingredient for preventing or ameliorating a prostate disease.

The present invention relates to a composition for preventing, ameliorating or treating a prostate disease, comprising an extract of Aspergillus oryzae as an active ingredient, wherein the extract of Nagoya mucilaginus, an active ingredient of the present invention, contains the weight of the enlarged prostate by the prostate hypertrophy And inhibit the expression of androgenic receptors or prostate specific antigen proteins, as well as significantly reduce the levels of testosterone and DHT in serum and prostate, so that the prostatic hyperplasia, prostatitis or prostate cancer Of prostate diseases.

Fig. 1 shows the results of confirming cell survival rate (%) according to the treatment of mulberry extract of the present invention. NC is a negative control with no treatment.
FIG. 2 is a Western blotting result of confirming the inhibitory effect of the extracts of androgenic receptor (AR) and prostate-specific antigen (PSA) proteins on the mucus extract. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + EV induces benign prostatic hyperplasia and mucus extract treatment; 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 + EV induces benign prostatic hyperplasia and mucus extract treatment; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 4 shows the results of the weight change of the prostate gland according to the administration of the mildew extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + EV induces benign prostatic hyperplasia and mucus extract treatment; 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 of serum according to the administration of the mulberry extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + EV induces benign prostatic hyperplasia and mucus extract treatment; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 6 is a graph showing a change in the content of testosterone in serum according to the administration of the extract of Aspergillus oryzae of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + EV induces benign prostatic hyperplasia and mucus extract treatment; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.
FIG. 7 shows the results of H & E staining of prostate tissue according to the administration of the mulberry extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + EV induces benign prostatic hyperplasia and mucus extract treatment; 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 mulberry extract of the present invention. NC is a negative control; BPH is the cause of benign prostatic hyperplasia; BPH + EV induces benign prostatic hyperplasia and mucus extract treatment; BPH + F is a group of benign prostatic hyperplasia-induced and positive control, Finasteride.

The present invention is based on the finding that Eriochloa 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 mildew extract may be prepared by a method including, but not limited to, the following steps:

(1) extracting an extractant 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 nidokpyok, and more preferably 5 to 15 times the weight of the dried extract. 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 mite extract may inhibit the 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 mildew 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.

In addition, the present invention is also directed to a method of treating < RTI ID = villosa extract as an active ingredient for preventing or ameliorating a prostate disease.

The health functional food composition for preventing or ameliorating a prostate disease containing the mildew extract as an active ingredient may be prepared by any one of beverage, ring, tablet, capsule, and acid, And can be suitably produced according to a conventional method.

Examples of the food to which the mushroom extract of the present invention can be added include meat products, sausages, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream, , Tea, a drink, an alcoholic beverage, and a vitamin complex, and includes all the health functional foods in a 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  1. I too outpost , 037-022) Preparation of extract

Using a 99.9% (v / v) methanol grade (HPLC grade) extraction solvent from Korea Plant Extract Bank, 21.44 mg of mulberry extract obtained at an extraction yield of 8.8% was distributed and 214.4 μl Of DMSO at a concentration of 100 mg / ml and used for in vitro experiments.

In addition, 7.60 g of mulberry mulberry extract, which was extracted from Korean Plant Extract Bank at a yield of 10.11% using methanol (HPLC grade) of 99.9% (v / v) as an extraction solvent, It was used for animal experiments ( in vivo ).

Example  2. Cells treated with mulberry extract Of cell viability  Confirm

To examine the effects of the mulberry extract on LN cap cells, 1 × 10 ⁴ LN cap cells were plated on 96-well plates and cultured at 37 ° C. for 24 hours. Then, 50, 100, 200 占 퐂 / ml), and then 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, as shown in FIG. 1, it was confirmed that even when the mulberry extract was treated, the cell viability was not affected even when the mulberry extract was treated, and thus, there was little 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 6-well plates and cultured for 24 hours. Then, 100 nM of testosterone was treated in all wells except the control (control) 200 / / ml), and then cultured in a 5% CO ₂ environment at 37 캜 for 72 hours. As a positive control, 10 μM Finasteride was used.

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 100 μg / ml of mulberry extract to 200 μg / ml of finasteride, (Fig. 3A).

As shown in FIG. 3B, the expression of prostate-specific antigen (PSA) protein was decreased when the mulberry 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 control group, the 5α-reductase inhibitor, Finasteride, which is used as a treatment for benign prostatic hyperplasia, was administered at a dose of 10 mg / kg Lt; / 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. Oral administration of pinasteride (Finasteride), which is used as a herbal extract and treatment for benign prostatic hyperplasia, was administered orally for 4 weeks. After the administration of the drug, sacrificed rat prostate glands were sampled and weighed.

As a result, as shown in Fig. 4, BPH + EV, prostatic hyperplasia + Finasteride group: BPH + prostate hyperplasia + F), the weight of the prostate gland in the group that caused enlargement of the prostate gland was increased compared to that of the normal group, and the weight of the prostate gland significantly decreased in the group treated with the glandular extract Respectively.

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 supernatant and the serum was analyzed by 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 dihydrotestosterone (DHT) in the serum was statistically significantly decreased in the group administered with the mulberry 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, testosterone content in the serum decreased statistically significantly in the group administered with the mulberry extract.

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, it was confirmed that the height and number of epithelium cells were prominent in the prostatic hyperplasia-induced group, but the proliferation of epithelial cells was suppressed in the group treated with the mulberry extract.

Example  9. Serum ALT and AST  Measure

After the end of the experiment, the separated blood was centrifuged at 4,000 rpm for 20 minutes and the supernatant was taken to confirm the change of ALT and AST contents in the serum.

As a result, as shown in FIG. 8, there was no statistically significant change in the content of ALT and AST used as general liver toxicity (damage) index in both the group with hypertrophy induced prostate and the group treated with herb extract I did.

Claims (8)

A pharmaceutical composition for preventing or treating hyperplasia of prostate comprising Eriochloa villosa extract as an active ingredient. delete The method of claim 1 wherein the I cigarettes extraction solvent of the 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 for preventing or treating enlargement of the prostate gland according to claim 1, wherein the mildew extract is an inhibitor of the expression of an androgenic receptor or a prostate specific antigen protein. [Claim 2] The pharmaceutical composition according to claim 1, wherein the mildew extract has a reduced amount of DHT (dihydrotestosterone) or testosterone in the 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. Also disclosed is a health functional food composition for preventing or ameliorating hyperplasia of prostate comprising Eriochloa villosa extract as an active ingredient. 8. The method according to claim 7, wherein the Eriochloa villosa extract is prepared by any one of a beverage, a pill, a tablet, a capsule and a powdered preparation. Health functional food composition.

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