KR102293358B1 - Composition for Preventing or Treating Benign Prostatic Hyperplasia Comprising Supercritical Fluid Extraction of Quisqualis Indica - Google Patents

Abstract

The present invention relates to a composition for preventing or treating benign prostatic hyperplasia, and more particularly, to a pharmaceutical composition and a food composition for preventing or treating benign prostatic hyperplasia, which include a supercritical extract of tetrapods. According to the present invention, the supercritical extract of Quercus chinensis exhibits an effect of reducing PSA expression and prostate weight, so that a composition containing it can be usefully used for the treatment and prevention of benign prostatic hyperplasia.

Description

A composition for preventing or treating benign prostatic hyperplasia Comprising Supercritical Fluid Extraction of Quisqualis Indica, containing a supercritical extract of quasi-Gunja

The present invention relates to a composition for preventing or treating benign prostatic hyperplasia, and more particularly, to a pharmaceutical composition and a food composition for preventing or treating benign prostatic hyperplasia, which include a supercritical extract of tetrapods.

Benign prostatic hyperplasia (BPH) refers to a disease accompanied by symptoms of blockage or decrease in the flow of urine in the urethra by blocking the passage of urine from the lower bladder as the prostate enlarges. The prostate surrounds the tube (ureter) that moves urine from the bladder. Benign prostatic hyperplasia (BPH) is a disease that makes it difficult to urinate as the gland grows intensively in the area next to the urethra with age. An enlarged prostate obstructs the flow of urine out of the bladder, making the bladder work harder to empty, and as this problem worsens over time, it can cause problems with passing urine completely. It can also thicken the bladder wall, which can lead to muscle spasms.

Testosterone is deeply involved in the growth, development and pathological condition of the prostate through male hormone receptors in the prostate. Prostatic hypertrophy is known to be a disease that is greatly affected by male hormones. In recent years, herbal remedies have been emerging a lot to compensate for the side effects and disadvantages of the existing synthetic drug therapies for treating 　prostate and hypertrophy. As an example, Republic of Korea Patent Publication No. 10-1883065 discloses a pharmaceutical composition for preventing or treating enlarged prostate 　 comprising a mixed extract of ginseng, baekchul, baekbokryeong, dermis, licorice, ginger and jujube as an active ingredient.

On the other hand, Quisqualis indica (Quisqualis indica) is a tendril plant belonging to the family Quesqualis family. A plant native to Southeast Asia, it is cultivated in various countries because of its beautiful flowers and fragrant scent. White five-leaf flowers bloom from summer to autumn, then turn red. Four gentleman fruit ( Quisqualis Fructus ) is used as a neutralizing agent for nicotine addiction, as a roundworm extermination, as an insecticide for children, and has a weak scent and a slightly sweet taste.

Numerous references are referenced throughout this specification and citations thereof are indicated. The disclosures of the cited documents are incorporated herein by reference in their entirety to more clearly describe the content of the present invention and the level of the art to which the present invention pertains.

Republic of Korea Patent Publication No. 10-1883065

An object of the present invention is to provide a pharmaceutical composition for the prevention or treatment of benign prostatic hyperplasia, which contains the supercritical extract of Sagunja as an active ingredient as a natural medicine.

Another object of the present invention is to provide a food composition for preventing or improving prostatic hyperplasia containing the extract.

Other objects and advantages of the present invention will become more apparent from the following detailed description of the invention, claims and drawings.

One aspect of the present invention is the four gentlemen ( Quisqualis indica ) to provide a pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising a supercritical fluid extract as an active ingredient.

The present inventors have tried to develop substances that can effectively treat or prevent enlarged prostate and are safe for the human body, especially plant-derived substances. When a supercritical fluid is used as an extraction solvent, the present invention was completed by finding that there is a significant improvement in prostatic hypertrophy and a therapeutic effect than other solvent extracts.

A supercritical fluid is a substance at a temperature and pressure above a critical point, and can diffuse through a solid such as a gas and dissolve a substance such as a liquid. Also, close to the critical point, small changes in pressure or temperature result in large changes in density, so that many properties of supercritical fluids can be fine-tuned. Supercritical fluids have highly compressed gases and combine the properties of gases and liquids in interesting ways. The rapid expansion of the supercritical solution leads to the precipitation of finely divided solids.

Examples of the extraction solvent used to obtain the quadrilateral supercritical extract of the present invention include, but are not limited to, carbon dioxide, water, methane, ethane, propane, ethylene, propylene, methanol, ethanol, acetone, and the like. One or more supercritical fluids may be mixed, and a permanent gas (eg, N ₂ or H ₂ ) may be mixed.

The extraction solvent used in the present invention is preferably carbon dioxide and/or water, and most preferably carbon dioxide. CO ₂ is an extraction solvent for plants and does not leave toxic substances behind. The properties of supercritical carbon dioxide extraction can be extensively and precisely manipulated with subtle changes in pressure and temperature.

When carbon dioxide is used as the extraction solvent in the present invention, it may be mixed with a co-solvent such as water, ethanol and/or methanol. The extraction conditions of supercritical carbon dioxide are above the critical pressure of 74 bar and the critical temperature of 31° C., more specifically, a pressure of 74 to 1000 bar and a temperature of 31 to 100° C., a pressure of 74 to 900 bar, and a temperature of 31 to 90° C. conditions, a pressure of 74 to 800 bar and a temperature of 31 to 80° C., a pressure of 74 to 700 bar and a temperature of 31 to 70° C., a pressure of 74 to 600 bar and a temperature of 31 to 60° C., or 74 to 500 bar. Pressure and temperature conditions of 31 to 50° C. may be used, but are not necessarily limited thereto.

In one embodiment of the present invention, the tetrapod supercritical extract of the present invention is obtained by pulverizing commercially available tetrapods, making pure carbon dioxide into a supercritical state, flowing ethanol as a co-solvent to recover the extract, and drying it. can

In another embodiment of the present invention, the quasi-Gunja extract may be Quisqualis Fructus extract.

As used herein, the term 'extract' refers to not only the extraction result obtained by treating the supercritical fluid extraction solvent on the genus genus, but also the processed (eg, powdered) tetrapod that is formulated to be administered to animals including humans. meaning to include For example, the tetrapod extract used in the present invention is liquid, or It may be prepared in a powder state by additional processes such as vacuum distillation and freeze drying or spray drying.

In addition, the term 'extract' used in the present specification has the meaning commonly used as a crude extract in the art as described above, but in a broad sense also includes a fraction obtained by additionally fractionating the extract. That is, the tetrapod extract includes not only those obtained by using the above-described supercritical extraction solvent, but also those obtained by additionally applying a purification process thereto. For example, a fraction obtained by passing the extract through an ultrafiltration membrane having a constant molecular weight cut-off value, separation by various chromatography (prepared for separation according to size, charge, hydrophobicity or affinity), etc. The fraction obtained through the purification method is also included in the tetrapod extract of the present invention.

The supercritical extract of tetrapods according to the present invention may contain at least one fatty acid selected from the group consisting of myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, arachidic acid, eicosenic acid, behenic acid, and ligroseric acid. and preferably 10 to 150 mg/g of linoleic acid, It may contain 100 to 300 mg/g of palmitic acid and 200 to 500 mg/g of oleic acid.

As described above, the supercritical extract of the quadrupedal plant of the present invention has significantly improved effects in relation to reduction in prostate specific antigen (PSA) expression and reduction in prostate weight, compared to other solvent extracts.

The composition of the present invention comprising the above-mentioned quadrilateral supercritical extract as an active ingredient may be prepared as a pharmaceutical composition, in which case it may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers included in the composition of the present invention are commonly used in formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate. , microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, etc., but are limited thereto no. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

The pharmaceutical composition of the present invention may be administered orally or parenterally, and is preferably applied by oral administration.

A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration method, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate and reaction sensitivity of the patient. can be A typical dosage of the pharmaceutical composition of the present invention is in the range of 0.001-100 mg/kg for adults.

The pharmaceutical composition of the present invention is prepared in unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person of ordinary skill in the art to which the present invention pertains. or may be prepared by incorporation into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension, syrup, or emulsion in oil or aqueous medium, or may be in the form of an extract, powder, powder, granule, tablet or capsule, and may additionally include a dispersant or stabilizer.

In addition, the composition of the present invention may be provided as a food composition. In this case, as an active ingredient, not only the tetrapod extract, but also ingredients commonly added during food production, for example, may include proteins, carbohydrates, fats, nutrients, seasonings and flavoring agents. Examples of the above-mentioned carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As the flavoring agent, natural flavoring agents such as taumatine, stevia extract (eg, rebaudioside A, glycyrrhizin) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used. For example, when the food composition of the present invention is prepared as a drink, citric acid, high fructose, sugar, glucose, acetic acid, malic acid, fruit juice, licorice root extract, jujube extract, licorice extract, etc. can do it

The composition comprising the supercritical extract of the present invention as an active ingredient exhibits the effect of reducing the expression of PSA and the weight of the prostate, and thus can be usefully used for the treatment and prevention of benign prostatic hyperplasia.

1 shows the effect of reducing the expression of antiprostate specific antigen (PSA). Here, Fin means the finasteride administration group.
Figure 2 shows the change in prostate weight according to the treatment of the tetrapod extract in an enlarged prostate animal model.

Hereinafter, the present invention will be described in more detail through examples. These examples are only for illustrating the present invention in more detail, and it will be apparent to those of ordinary skill in the art to which the present invention pertains that the scope of the present invention is not limited by these examples.

Example

four gentlemen supercritical extract preparation

Commercially available four gentlemen ( Quisqualis) indica ) was purchased and pulverized to a uniform size and 2 kg was placed in a supercritical fluid extractor (10L). Then, using a liquid chromatography pump (supercritical fluid chromatograph) to make pure carbon dioxide in a supercritical state at a temperature of 40 ℃ and a pressure of 400 bar, and while flowing ethanol as a cosolvent, it was recovered in the flask for 10 hours. The ethanol of the supercritical extract was dried with a rotary vacuum evaporator, and was obtained in a yield of 9.71% with a dry weight of 194.2 g.

Manufacture of Four Gentleman's Alcoholic Extracts

Commercially available four gentlemen ( Quisqualis) indica ) was purchased and pulverized, and 300 g was added to 3 L of 70% alcohol and ultrasonically extracted three times for 1 hour using an ultrasonic extractor. The extract was mixed with Whatman No. After filtration with 2 (150 mm Φ) filter paper to remove insoluble substances, the mixture was 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, 500 mL of purified water was added and suspended, and then 80.51 g of the extract was obtained using a freeze dryer (yield: 26.84%).

four gentlemen fraction Produce

Dissolve 50 g of the prepared sagebrush extract in 500 ml of purified water, add 500 ml of BuOH, stir at 500-800 rpm for 1 hour, and repeat twice to obtain BuOH and water layers, respectively, and Whatman No. After filtration with 2 (150 mm Φ) filter paper to remove insoluble substances, the fractions were obtained by concentration under reduced pressure at 40° C. with a concentrator equipped with a cooling condenser to obtain fractions each of 8 g of BuOH and 40 g of water.

four gentlemen supercritical Fatty Acid Analysis of Extracts

The fatty acid composition of the supercritical and alcohol extracts of Sagunja supercritical and alcohol was dissolved in heptane and then converted into fatty acid methyl ester through transesterification with a methanol solution in which 2N concentration of KOH was dissolved. It was confirmed through FID analysis. As the analysis equipment, an American Agilent 7890A model was used as GC/FID, and DB-WAX (30m>0.25mm>0.25㎛, J&W Scientific, USA) was used as the analysis column. At this time, the temperature of the oven was maintained at 100° C. for 2 minutes, then heated to 200° C. at a rate of 25° C./min, and up to 230° C. at a rate of 5° C./min, followed by maintaining the temperature for 35 minutes for analysis. Hydrogen gas was used as the mobile phase, and 1 μl of the sample was injected, and the split ratio was set to 50:1. In addition, the temperature of the injection part and the temperature of the detector were maintained at 250 °C.

As such, the results of comparing the fatty acids of the supercritical and alcohol extracts of Sagunja are shown in Table 1 below.

compound name Four Gentleman Alcoholic Extract (mg/g) Four Gentleman Supercritical Extract (mg/g) Myristic acid 0.252 48.076 Palmitic acid 2.403 231.761 Stearic acid 0.24 24.989 Elaidic acid 1.644 _ Oleic acid 4.6446 431.135 Linoleic acid 2.043 82.994 Arachidic acid 0.054 2.13 Eicosenoic acid 0.03 2.381 Behenic acid 0.055 1.148 Lignoceric acid 0.042 0.732

Prostate specific antigen ( Antiprostate specific antigen; PSA) measurement

Prostate cancer cells (human prostate cancer cell line; LNCaP) were treated with the same concentration of each supercritical, alcohol extract, BuOH, and water fraction and cultured for 72 hours. After that, protein was extracted by applying RIPA lysis buffer to LNCaP cells, and 30-40 ug of protein was separated from 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel, and then electrophoresed at 40V for 2 hours. It was transferred to a nitrocellulose membrane by electrophoresis. The expression level of prostate-specific antigen (PSA) protein among the isolated proteins was evaluated, and the results are shown in FIG. 1 .

As a result of the experiment, the superior PSA expression inhibitory effect was shown in the quasi-dragon extract and the supercritical extract of the quinceanium compared to the fractions for each solvent. BuOH and water, which are fractions compared to the quasi-drinking alcohol extract, had a mild effect and had low industrial and economic value, so the effect of supercritical and alcohol extracts on prostate weight reduction was evaluated.

Prostate weight reduction effect

(1) animal model

After acclimatization of 10-week-old male Wister rats (central laboratory animal) for 1 week, the prostate enlargement induction group was subcutaneously injected with 3 mg/kg of testosterone propionate (TP) for 4 weeks to make a model. An hour before the injection of TP, the supercritical and alcohol extracts of Sagunja were orally administered at 300 mg/kg for 4 weeks, and as a positive control, Carito (Carito ^® ; 300 mg/kg), which is used as a treatment for benign prostatic hyperplasia, was administered in the same way. did.

(2) Prostate weighing

After sacrificing the animals, each group (normal group; NC, BPH induced group; BPH, BPH induced group + Carito administration group; BPH+Carito ^® , BPH induced group + Quagunja supercritical extract group; BPH + supercritical, prostate Prostates were extracted from the hypertrophy-inducing group + group administered with aloe vera extract; BPH + sagunja) and their weight was measured.

(3) Statistical analysis

Statistical analysis was performed by ANOVA, and ##P<0.01, #P<0.05 was significantly different from the normal group (NC), **P<0.01, *P<0.05 caused BPH A case with a significant difference compared with the group (BPH) was considered.

(4) Experiment result

The results of the prostate weight measurement and statistical analysis are shown in FIG. 2 . As a result of the experiment, it was confirmed that the weight of the prostate was significantly reduced in the group (BPH+supercritical) administered with the tetrapods supercritical extract than in the group (BPH) induced by TP hypertrophy. It exhibited similar effects to Carito, which is currently used as a treatment for benign prostatic hyperplasia, and exhibited a superior effect than the Sagunja alcohol extract.

Claims (9)

Four gentlemen ( Quisqualis) indica ) A pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising a supercritical fluid extract as an active ingredient. The composition of claim 1 , wherein the fluid comprises carbon dioxide. The composition according to claim 1, wherein the supercritical fluid extract of the quadrilaterals is extracted using supercritical carbon dioxide at a pressure of 74 to 1000 bar and a temperature of 31 to 100°C. The composition of claim 1, wherein the composition has an effect of reducing the expression of prostate specific antigen (PSA). The method of claim 1, wherein the extract comprises one or more fatty acids selected from the group consisting of myristic acid, palmitic acid, stearic acid, oleic acid, linoleic acid, arachidic acid, eicosenic acid, behenic acid, and ligroceric acid. composition. According to claim 1, wherein the extract is linoleic acid 10 to 150 mg / g, A composition comprising 100 to 300 mg/g of palmitic acid and 200 to 500 mg/g of oleic acid. The composition of claim 1, wherein the extract is a liquid or dry powder. The composition of claim 1, wherein the composition is for oral administration. Four gentlemen ( Quisqualis) indica ) A food composition for preventing or improving prostatic hyperplasia comprising a supercritical extract as an active ingredient.

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