KR102146567B1 - Composition for preventing, ameliorating or treating prostate disease comprising Dianthus chinensis extract as effective component - Google Patents

Abstract

The present invention relates to a composition for preventing, improving or treating prostate disease, comprising a dianthus plant extract as an active ingredient. The dianthus plant extract, which is an active ingredient of the present invention, reduces the weight of the prostate thickened by prostate hypertrophy (BPH), and can significantly reduce the content of testosterone and dihydrotestosterone (DHT) in serum and prostate. Therefore, the composition of the present invention can be used as a health functional food or medicine for the prevention, improvement or treatment of prostatic hyperplasia.

Description

Composition for preventing, ameliorating or treating prostate disease comprising Dianthus chinensis extract as effective component, comprising a dianthus plant extract as an active ingredient

The present invention relates to a composition for preventing, improving or treating prostate disease, comprising a dianthus plant extract as an active ingredient.

The prostate gland is an organ that only men have, and it is located under the bladder and is in the form of an upside down chestnut. The prostate is so small that it is difficult to detect immediately after birth, but it gradually becomes larger by the action of male hormones as puberty becomes. As can be seen from the etymology of pro(前) + state(立) + gland(腺), the prostate refers to an organ of glandular tissue located in the front part of the lower abdomen of the human body. In other words, the prostate is an organ consisting of a glandular tissue that secretes a substance and a fibrom uscular tissue that surrounds the prostate. It is an institution.

Prostate secretion fluid is opened in the prostate urethra area along 15 to 30 tubes. The prostate begins to grow after puberty under the influence of male hormones, and becomes about 20 mg in size around 30 years of age. When the prostate is over 60 years old, it is a type of aging phenomenon, which generally increases in size, leading to prostatic hypertrophy. However, even if there is an enlarged prostate, it does not necessarily mean that treatment is necessary, and how uncomfortable and suffering a patient feels due to this is the first application criterion for treatment.

Benign prostatic hyperplasia (BPH) was defined as a condition in which the flow of urine decreased due to obstruction of the urethra by blocking the passage of urine from the lower part of the bladder due to an enlarged prostate. Histologically, prostate epithelial cells or prostate cells Was defined as proliferated. In recent years, the pathophysiology of the disease is too complicated to explain with such a definition or concept, and now'in men over 50 years of age, they urinate more than 8 times a day, urinary frequency, nocturia, and strong and sudden urination (feeling peeing) When urinating while urinating is difficult, bladder storage symptoms such as unbearable urgency, delayed urine (a phenomenon in which urine comes out only when the urine is heated), severing urine (a phenomenon in which the flow of urine is interrupted), and a phenomenon in which you need to give strength during urination Prostate enlargement is defined as'complaining of lower urinary tract symptoms, collectively referred to as symptoms indicating bladder drainage disorder'.

The main clinical features are signs of prostate enlargement and lower urinary tract. It is known that prostate enlargement occurs in the presence of androgens, and anabolic steroids increase the capacity of the prostate and decrease urine flow, causing an increased urinary frequency.

The prostate is an androgen-dependent organ in which testosterone and its extratesticular origin are activated with a more potent dihydrotestosterone (DHT). Typically, the prostate is an important organ of DHT formation, and the systemic effect of the prostate's endocrine activity is mainly involved in DHT formation and its excretion for circulation. The production of DHT increases with age, leading to increased prostate growth and hypertrophy. The importance of DHT is confirmed by clinical studies in which 5α-reductase inhibitors have been applied to male patients with BPH. In many cases, it is known that treatment with 5α-reductase inhibitors significantly reduces the DHT levels and prostate size in the prostate. Finasteride is widely used in the treatment of androgen-dependent diseases such as male pattern baldness, enlarged prostate (BPH) and prostate cancer. Finasteride is a competitive and specific inhibitor of 5α-reductase, blocking DHT conversion of testosterone in the prostate, hair follicles and other androgen-sensitive tissues, resulting in inhibition of DHT concentrations in serum and prostate. Conventionally used drugs, such as finasteride and dutasteride, have proven to be effective treatment methods for BPH, but their use is erectile dysfunction, loss of libido, dizziness and the above. It is strictly limited due to side effects such as upper respiratory tract infection.

Meanwhile, Dianthus chinensis L.) is a perennial plant in the family stone bamboo family. The flowers are bisexual, bloom in June to August, hang one at the end of the branch, and are red. Also known as Seokjukhwa, Daeran, and Sangumaek. It grows in dry places in lowlands or in sandy lands by streams. The stems are densely gathered, the height is around 30cm, and the branches are split from the top. The leaves are opposite and merge at the bottom to surround the main stem, and the edges are flat in a row shape. The flowers are bisexual, bloom in June to August, hang one at the end of the branch, and are red. The calyx is divided into 5 and the bottom is cylindrical. The number of petals is 5, the bottom part is thinner, the front part is spread to the side, the tip is split shallowly, and there is a dark pattern. There are 10 stamens and 2 styles. Fruits are capsules, wrapped with calyx, split into 4 at the end, and ripen in September to October. There are 4-6 bracts below. The whole with flowers and fruits is dried in the shade and used as a medicinal material, and it is used for diuresis, pain, anti-inflammatory, etc. It is distributed in Korea and China. Dianthus (var. semperflorens) blooms year-round. The color is dark red and there are many horticultural varieties. In Japan, it is also called tokonatsu, which means that it is always summer because flowers continue to bloom.

As a prior art known in relation to the present invention, a composition for preventing and treating prostatic hyperplasia is disclosed in Korean Patent No. 1165719 containing Janggu-chae extract as an active ingredient, and alpha-spinasterol is disclosed in Korean Patent No. A pharmaceutical composition for preventing or treating androgen-related diseases, diseases or conditions containing as an active ingredient is disclosed, but mentions a composition for preventing, improving or treating prostate diseases comprising the dianthus plant extract of the present invention as an active ingredient Never happened.

The present invention is derived from the above requirements, the present invention provides a composition for the prevention, improvement or treatment of prostate disease comprising a dianthus plant extract as an active ingredient, wherein the dianthus plant extract is testosterone in blood and prostate tissue. And by confirming that it effectively reduces the content of dihydrotestosterone (DHT), the present invention was completed.

In order to achieve the above object, the present invention provides a pharmaceutical composition for the prevention or treatment of prostate disease containing a dianthus plant extract as an active ingredient.

In addition, the present invention provides a health functional food composition for preventing or improving prostate disease, comprising a dianthus flower plant extract as an active ingredient.

The present invention relates to a composition for preventing, improving or treating prostate disease, comprising a dianthus plant extract as an active ingredient. The dianthus plant extract, which is an active ingredient of the present invention, reduces the weight of the prostate thickened by prostate hypertrophy (BPH), and can significantly reduce the content of testosterone and dihydrotestosterone (DHT) in serum and prostate. Therefore, the composition of the present invention can be effectively applied to the prevention, improvement or treatment of prostatic hyperplasia, and since the dianthus plant extract is a natural product derived from a plant, it is excellent in safety and can be very advantageously applied to medicines, quasi-drugs and foods.

1 is a result of measuring the prostate weight of the rat to which the dianthus plant extract of the present invention was administered, NC as a normal group, a group in which PBS was orally administered while subcutaneous injection of corn oil, and BPH in order to induce prostatic hyperplasia TP (testosterone propionate) was administered subcutaneously and PBS was orally administered.Fin was orally administered finasteride during TP subcutaneous injection, and dianthus plant extract was orally administered with dianthus plant extract during TP subcutaneous injection. It is one group. ## indicates that the prostate weight of the BPH group was statistically significantly increased compared to the normal group (NC), p<0.01, and *, ** indicates the group administered with dianthus plant extract or finasteride compared to the BPH group. It means that the prostate weight of the administered group was statistically significantly reduced, * = p<0.05, ** = p<0.01.
Figure 2 is a result of confirming the effect of reducing the content of dihydrotestosterone (DHT) (A) and testosterone (Testosterone; TS) (B) in the prostate tissue according to administration of a dianthus plant extract. NC is a normal group, which is a group in which PBS is orally administered while subcutaneous injection of corn oil, BPH is a group in which TP (testosterone propionate) is injected subcutaneously to induce prostatic hyperplasia, while PBS is orally administered, and Fin is TP. During the subcutaneous injection period, finasteride was orally administered, and the dianthus plant extract was orally administered with the dianthus plant extract during the TP subcutaneous injection period. ## means that the content of DHT (A) or testosterone (B) in the prostate tissue of the BPH group was statistically significantly increased compared to that of the normal group (NC), and p<0.01, ** is compared with the BPH group. Thus, the content of DHT (A) and TS (B) in the prostate tissue of the group to which the dianthus plant extract was administered orally was statistically significantly reduced, p<0.01.
Figure 3 is a result of confirming the effect of reducing the content of dihydrotestosterone (DHT) (A) and testosterone (Testosterone; TS) (B) in serum according to the administration of a dianthus plant extract. NC is a normal group, and PBS is orally administered while subcutaneous injection of corn oil, BPH is a group that orally administered PBS while subcutaneous injection of TP (testosterone propionate) to induce prostatic hyperplasia, and Fin is TP subcutaneous During the injection period, finasteride was orally administered, and the dianthus plant extract was a group administered with the dianthus plant extract orally during the TP subcutaneous injection period. ## means that the content of DHT (A) and TS (B) in the serum of the BPH group was statistically significantly increased compared to that of the normal group (NC), p<0.01, *, ** are the BPH group In contrast, it means that the content of DHT (A) and testosterone (B) in the serum of the group administered finasteride was statistically significantly reduced, * = p<0.05, ** = p<0.01.
4 is a histological analysis result confirming the effect of a dianthus plant extract on prostatic hyperplasia, and is a photograph of H&E stained prostate tissue (magnification × 200). NC is a normal group, and PBS is orally administered while subcutaneous injection of corn oil, BPH is a group that orally administered PBS while subcutaneous injection of TP (testosterone propionate) to induce prostatic hyperplasia, and Fin is TP subcutaneous During the injection period, finasteride was orally administered, and the dianthus plant extract was a group administered with the dianthus plant extract orally during the TP subcutaneous injection period.

The present invention relates to a pharmaceutical composition for the prevention or treatment of prostate disease containing a dianthus plant extract as an active ingredient.

The dianthus plant extract may be prepared by a method comprising the following steps, but is not limited thereto:

(1) extracting by adding an extraction solvent to the dianthus plant;

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

(3) preparing an extract by concentrating the filtered extract in step (2) under reduced pressure and drying it.

In the step (1), the extraction solvent is preferably selected from water, a lower alcohol of C ₁ to C ₄ or a mixture thereof, more preferably a lower alcohol of C ₁ to C ₄ , and even more preferably 70% (v/v) ethanol, but is not limited thereto.

In the above manufacturing method, all conventional methods known in the art, such as filtration, hot water extraction, immersion extraction, reflux cooling extraction, and ultrasonic extraction, can be used as the extraction method. The extraction solvent is preferably extracted by adding 1 to 20 times the weight of the dried dianthus plant, more preferably 5 to 15 times. The extraction temperature is preferably 4 to 50 ℃, but is not limited thereto. In addition, the extraction time is preferably 0.5 to 10 hours, more preferably 0.5 to 5 hours, but is not limited thereto. In the above method, the vacuum concentration in step (3) is preferably a vacuum vacuum concentrator or a vacuum rotary evaporator, but is not limited thereto. In addition, the drying is preferably vacuum drying, vacuum drying, boiling drying, spray drying, or freeze drying, but is not limited thereto.

The term'extract' used in the present invention has a meaning commonly used as a crude extract in the art, but broadly includes a fraction obtained by further fractionating the extract. That is, the dianthus flower plant extract includes not only those obtained by using the above-described extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, fractions obtained by passing the extract through an ultrafiltration membrane having a certain molecular weight cut-off value, separation by various chromatography (one prepared for separation according to size, charge, hydrophobicity or affinity), etc. Fractions obtained through various purification methods are also included in the dianthus plant extract of the present invention.

The prostate disease is preferably, but not limited to, benign prostatic hyperplasia (BPH).

The composition of the present invention reduces the content of testosterone or DHT (dihydrotestosterone) in blood or tissue, and the blood is preferably serum, but is not limited thereto, and the tissue is prostate, hair follicles, or Other androgen-sensitive tissues (eg, testes, kidneys, bones, etc.).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and when administered parenterally, it is preferable to select an injection method for external use of the skin or intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine dural or cerebrovascular injection.

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 is the type of disease, severity, and drug activity of the patient. , Sensitivity to drugs, time of administration, route of administration and rate of excretion, duration of treatment, factors including drugs used concurrently, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or administered in combination with other therapeutic agents, may be administered sequentially or simultaneously with a conventional therapeutic agent, and may be administered single or multiple. It is important to administer an amount capable of obtaining the maximum effect in a minimum amount without side effects in consideration of all the above factors, and this can be easily determined by a person skilled in the art.

The dosage of the pharmaceutical composition of the present invention can be used in various ranges according to the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and severity of disease.

The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable carrier, excipient, or diluent in addition to the active ingredient, the dianthus plant extract.

The pharmaceutical composition of the present invention may be in various oral or parenteral dosage forms. In the case of formulation, it is prepared using diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants that are usually used. Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and such solid preparations include at least one excipient in one or more compounds, such as starch, calcium carbonate, sucrose, or lactose ( lactose), gelatin, etc. In addition, in addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Liquid preparations for oral administration include suspensions, liquid solutions, emulsions, syrups, etc.In addition to water and liquid paraffin, which are commonly used simple diluents, various excipients such as wetting agents, sweetening agents, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterilized aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized agents, and suppositories. As the non-aqueous solvent and the suspension solvent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate may be used. As a base for suppositories, witepsol, macrogol, tween 61, cacao butter, laurin paper, glycerol gelatin, and the like may be used.

In addition, the present invention relates to a health functional food composition for preventing or improving prostate disease containing a dianthus plant extract as an active ingredient.

The health functional food composition of the present invention may be prepared by any one selected from beverages, pills, tablets, capsules, and powders, or may be prepared by adding them to food or food ingredients, and is appropriate according to a conventional method. Can be manufactured.

Examples of foods to which the dianthus plant extract of the present invention can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, and beverages. , Tea, drink, alcoholic beverage and vitamin complex may be in any one form selected from, and includes all health food in the usual sense.

The health functional food includes various nutrients, vitamins, minerals (electrolytes), synthetic and natural flavoring agents, coloring agents and enhancers (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, It may contain pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, and the like. In addition, it may contain pulp for the manufacture of natural fruit juices and vegetable beverages. These components may be used independently or in combination.

The health functional food of the present invention may contain various flavoring agents or natural carbohydrates as additional ingredients. The natural carbohydrates are monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the sweetener, natural sweeteners such as taumatin and stevia extract, and synthetic sweeteners such as saccharin and aspartame can be used.

Hereinafter, the present invention will be described in more detail using examples. These examples are only for describing the present invention in more detail, and it is apparent to those of ordinary skill in the art that the scope of the present invention is not limited thereto.

1. Preparation of Animal Model

200-220 g of male SD rats (Sprague-Dawley rats, Orient Bio) were acclimatized for 1 week, and then 6 rats per group were used for the experiment. Breeding was reared in a breeding room maintaining a 12-hour light/dark cycle at a temperature of 18-23°C under 40-60% relative humidity, and the rats were managed with a standard laboratory diet containing any water. All experimental procedures were conducted in accordance with the NIH guidelines for the management and use of experimental animals, and animal handling was conducted in accordance with the National Animal Welfare Law of Korea.

2. Induction and treatment of prostatic hyperplasia (BPH)

Rats were randomly classified into 5 groups after a week of acclimatization (n=6). The control group in which benign prostatic hyperplasia (BPH) was induced was prepared by subcutaneous injection of 10 mg/kg of TP (testosterone propionate) dissolved in corn oil daily for 4 weeks to induce prostatic hyperplasia, and the experimental group was prepared with TP for 4 weeks. One hour before the subcutaneous injection, 100mg/kg of dianthus plant extract was orally administered.

On the other hand, the normal control group was injected subcutaneously with corn oil instead of the dianthus plant extract, and the positive control group was 10 mg/kg of finasteride, a 5α-reductase inhibitor, which is used as a treatment for prostatic hyperplasia. It was administered orally.

The effective dosage of finasteride was based on previous reports, and the body weight of the animal model was measured weekly during the experiment. The application volume was pre-calculated based on the most recent body weight of the individual animals. After the final treatment, all animals that were fasted overnight were anesthetized with pentobarbital. Blood samples were obtained from the caudal vena cava. Serum was obtained through centrifugation and stored at 80°C. The total prostate weight (g) was checked immediately after removal.

For histological analysis, a part of the ventral prostate lobe was fixed with 10% formalin and embedded in paraffin. The rest of the prostate was stored at -80°C for measurement of hormone levels.

3. Homogenate Produce

Prostate tissue was homogenized in tissue lysis/extraction reagent (1/10 w/v) containing protease inhibitors (Sigma, MI, USA) using a homogenizer. The homogenate was centrifuged at 12,000 g for 25 minutes at 4° C., and the protein concentration in the supernatant fraction was measured using Bradford reagent (Bio-Rad, Hercules, CA).

4. Testosterone (TS) and Of dihydrotestosterone (DHT) Analysis of changes in serum and prostate content

The levels of testosterone and DHT in serum and prostate tissue were measured by enzyme-linked immunosorbent assay (ELISA). The DHT kit was purchased from ALPCO Diagnostics (Salem, NH) and the testosterone (TS) kit was purchased from Cayman (MI, USA).

5. Histopathological examination ( Histopathological examination)

To investigate the morphological changes in the prostate tissue, the tissue embedded in paraffin was cut into 4 μm-thick sections and stained with H&E solution (hematoxylin, Sigma MHS-16; and eosin, Sigma HT110-1-32). The tissues were mounted on a cover slide with Dako mounting medium (Dakocytomation, Denmark, CA) and observed under a microscope.

6. Statistical Analysis

Data were expressed as mean value ± standard deviation (SD). Statistical significance was determined through Student two-tailed t-test comparing independent mean values, and the statistical significance level was set to p<0.05.

Example 1. Preparation of dianthus plant extract

After pulverizing 300 g of dried dianthus, 3 L of 70% (v/v) ethanol was added thereto, followed by ultrasonic extraction 3 times for 1 hour at room temperature. The extract was filtered under reduced pressure with Advantec No. 2 (110mm) filter paper to remove insoluble substances and then concentrated under reduced pressure at 40°C using a concentrator equipped with a cooling condenser. In order to completely remove the solvent of the extract concentrated under reduced pressure, 0.5 L of purified water was added and suspended, and then 56.89 g (yield 18.96%) of dianthus ethanol extract was obtained using a freeze dryer.

Example 2. Confirmation of effect of reducing prostate weight by administration of dianthus plant extract

The relative prostate weight of TP-induced BPH animals was significantly higher compared to that of the normal group (NC). Compared with the TP-induced BPH group, the relative prostate weight (g) was significantly reduced in the group administered with the dianthus plant extract (FIG. 1 ).

Example 3. Dihydrotestosterone in prostate tissue following administration of dianthus plant extract ( Dihydrotestosterone ; DHT ) And testosterone ( Testosteron ; TS) content reduction effect confirmation

TP-induced BPH animals showed a significantly higher content of DHT in prostate tissue compared to the normal group (NC), but animals administered with dianthus plant extract were compared with TP-induced BPH animals. DHT content was found to be a statistically significant level of low content (Fig. 2A).

In addition, the content of testosterone (TS) in the prostate tissue was also lower than that of the TP-induced BPH animal group like DHT (FIG. 2B).

Example 4. In serum following administration of dianthus plant extract DHT And check the effect of reducing the content of testosterone (TS)

TP-induced BPH animals showed significantly higher serum DHT content compared to normal group (NC), but animals administered with dianthus plant extract showed DHT content in serum compared to TP-induced BPH animals. This showed a statistically significant low content (Fig. 3A).

In addition, the content of testosterone (TS) in serum also showed a statistically significant lower content compared to the TP-induced BPH animal group like DHT (FIG. 3B).

Example 5. Histological analysis of prostate tissue following administration of dianthus plant extract

The epithelial cell layer of the prostate was thicker in the TP-induced BPH group than in the normal group (NC), and in the group to which the dianthus plant extract was administered, the thickness was almost similar to that of the normal group (FIG. 4).

Claims (7)

A pharmaceutical composition for the prevention or treatment of prostatic hyperplasia (BPH) containing a dianthus flower plant extract as an active ingredient. The pharmaceutical composition for preventing or treating prostatic hyperplasia according to claim 1, wherein the extraction solvent of the dianthus plant extract is water, a lower alcohol of C ₁ to C ₄ or a mixture thereof. delete The pharmaceutical composition for preventing or treating prostatic hyperplasia according to claim 1, wherein the dianthus plant extract reduces the content of testosterone or dihydrotestosterone (DHT) in blood or tissue. The pharmaceutical composition for preventing or treating prostatic hyperplasia according to claim 1, further comprising a carrier, an excipient, or a diluent in addition to the dianthus plant extract. A health functional food composition for preventing or improving prostatic hyperplasia, comprising a dianthus flower plant extract as an active ingredient. The health functional food composition of claim 6, wherein the dianthus plant extract is prepared in any one formulation selected from beverages, pills, tablets, capsules, and powders. .

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