KR102556622B1 - Composition for preventing or treating benign prostatic hyperplasia comprising extracts of Panax ginseng, Glycyrrhiza uralensis and Paeonia lactiflora - Google Patents

Abstract

The present invention relates to a pharmaceutical composition and a food composition for the prevention, improvement or treatment of prostatic hyperplasia comprising extracts of ginseng, licorice and peony as active ingredients, wherein the mixed extract is a single extract of ginseng, licorice and peony of the present invention and These mixed extracts have excellent 5-alpha-reductase expression inhibitory effect and prostatic hyperplasia inhibitory effect, alleviate the side effects of finasteride, which has been used for the treatment of benign prostatic hyperplasia, and do not affect sperm count. Since it exhibits an effective therapeutic effect as a therapeutic agent, it can be usefully used in related industries.

Description

Composition for preventing or treating benign prostatic hyperplasia comprising extracts of Panax ginseng, Glycyrrhiza uralensis and Paeonia lactiflora

The present invention relates to a composition for preventing or treating benign prostatic hyperplasia comprising ginseng, peony and licorice extracts as active ingredients.

Benign prostatic hyperplasia (BPH) was defined as a condition in which the prostate enlarged and blocked the urinary tract at the lower part of the bladder, reducing the flow of urine. Indicating bladder emptying disorders, such as urinary incontinence (strong, sudden need to urinate), urgency (difficulty urinating), retardation (urinary output is delayed), interrupted urination (urine flow is interrupted), or straining during urination Prostatic hyperplasia is defined as 'complaints of lower urinary tract symptoms'.

Age and male hormones are considered the most important triggers for the occurrence of BPH. Histological enlargement of the prostate begins at the age of 35, and enlargement of the prostate appears in 60% of men in their 60s and 90% of men in their 80s. , 50% of these patients complained of dysuria due to prostate enlargement.

Drug treatment for BPH is largely classified into alpha blockers and androgen inhibitors (5 alpha-reductase inhibitors). Alpha-blockers are drugs that lower pressure and tension in the prostate and urethra. Commercially available alpha-blockers include terazosin, doxazosin, tamsulosin, and alfuzosin. However, alpha-blockers are only a temporary treatment method because they have no effect on reducing the enlargement of the prostate, and side effects such as dizziness, lethargy, headache, and visual disturbance have been reported. Androgen inhibitors are drugs that prevent enlargement of the prostate by inhibiting 5-alpha-reductase, which plays a key role in the process of enlargement of the prostate. Finasteride and dutasteride are commercially available as 5-alpha-reductase inhibitors. However, these drugs take a long time to take effect after taking them, and side effects related to sexual function have been reported.

As such, since most of the drugs currently on the market or under development exhibit various side effects, it is necessary to develop a new mechanism of treatment for benign prostatic hyperplasia.

In order to solve the above problems, the present inventors confirmed that ginseng, licorice and peony extracts have an effect of inhibiting prostate enlargement, and completed the present invention.

An object of the present invention is to provide a pharmaceutical composition for preventing or treating prostatic hyperplasia.

Another object of the present invention is to provide a health functional food for preventing or improving prostatic hyperplasia.

Another object of the present invention is to provide a method for preventing or treating prostatic hyperplasia.

In order to achieve the above purpose,

The present invention provides a pharmaceutical composition for preventing or treating prostatic hyperplasia, comprising at least one extract selected from the group consisting of ginseng ( Panax ginseng ), licorice ( Glycyrrhiza uralensis ) and peony ( Paeonia lactiflora ) as an active ingredient.

In one embodiment of the present invention, the extract may be a single extract of ginseng, licorice and peony, or a mixed extract of ginseng, licorice and peony.

In one embodiment of the present invention, the mixed extract may be extracted by mixing ginseng, licorice and peony at a weight ratio of 0.1 to 10:0.1 to 10:0.1 to 10, preferably, 0.5 to 3:0.5 to It may be extracted by mixing at a weight ratio of 3:0.5 to 3, more preferably, it may be extracted by mixing at a weight ratio of 1.1 to 3:1:1 or 1:1.1 to 3:1.

In addition, the present invention provides a food composition for preventing or improving prostatic hyperplasia comprising at least one extract selected from the group consisting of ginseng ( Panax ginseng ), licorice ( Glycyrrhiza uralensis ) and peony ( Paeonia lactiflora ) as an active ingredient. .

Furthermore, the present invention provides a method for preventing or treating prostatic hyperplasia comprising administering to a patient the pharmaceutical composition for preventing or treating BPH according to the present invention.

The present invention applies ginseng, licorice, and peony extracts, which are biocompatible and have no side effects as natural product-derived ingredients, to the prevention or treatment of prostatic hyperplasia. Expression suppression effect and BPH inhibitory effect were confirmed, and it was confirmed that it did not affect sperm count by alleviating the side effects of finasteride, a previously used treatment for BPH. Therefore, the extracts of ginseng, licorice and peony of the present invention and their mixed extracts show effective therapeutic effects as a treatment for prostatic hyperplasia, and thus can be usefully used in related industries.

1 shows the treatment of ginseng, licorice and peony extracts alone or mixed extracts (ginseng: licorice: peony = 1:1:1, 2:1:1, 1:2:1 and 1:1:2) according to the present invention. It is a graph showing the cell viability of human prostate epithelial cells (RWPE-1 cells).
Figure 2 is a single extract or mixed extract of ginseng, licorice and peony of the present invention (ginseng: licorice: peony = 1: 1: 1, 2: 1: 1, 1: 2: 1 and 1: 1: 2) according to the treatment It is a graph showing the result of analyzing the expression level of 5-alpha-reductase in human prostate epithelial cells through RT-PCR.
Figure 3 is a graph showing the results of the prostate index correcting the weight of prostate tissue for the weight and weight of prostate tissue in each group ( NC group : normal control group; BPH group : disease control group; ginseng group : ginseng extract injected experimental group; and Fi group : positive control group injected with finasteride).
Figure 4 is a graph showing the results of the prostate index correcting the weight of prostate tissue for the weight and weight of prostate tissue in each group ( NC group : normal control group; BPH group : disease control group; licorice group : licorice extract injected Experimental group; Fi group : positive control group injected with finasteride; and licorice+Fi group : licorice extract and finasteride combined treatment group).
Figure 5 is a graph showing the results of the prostate index correcting the weight of prostate tissue for the weight and weight of prostate tissue in each group ( NC group : normal control group; BPH group : disease control group; peony group : peony extract injected experimental group; and Fi group : positive control group injected with finasteride).
6 is a graph showing the results of counting and comparing the number of sperm in the ginseng extract-injected group and the finasteride-injected group in the epididymis ( NC group : normal control group; BPH group : disease control group; Gi group : ginseng extract-injected experimental group; Fi group : positive control; and Fi+Gi group: ginseng extract and finasteride combined treatment group).

Hereinafter, embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the following description, detailed descriptions of well-known technologies to those skilled in the art may be omitted. In addition, in describing the present invention, if it is determined that a detailed description of a related known function or configuration may unnecessarily obscure the gist of the present invention, the detailed description may be omitted. In addition, the terms used in this specification (terminology) are terms used to appropriately express preferred embodiments of the present invention, which may vary according to the intention of a user or operator or customs in the field to which the present invention belongs.

Therefore, definitions of these terms should be made based on the contents throughout this specification. Throughout the specification, when a certain component is said to "include", it means that it may further include other components without excluding other components unless otherwise stated.

The present invention provides a pharmaceutical composition for preventing or treating prostatic hyperplasia, comprising at least one extract selected from the group consisting of ginseng ( Panax ginseng ), licorice ( Glycyrrhiza uralensis ) and peony ( Paeonia lactiflora ) as an active ingredient.

In the present invention, the ginseng (人蔘, Ginseng Radix) is the root of ginseng ( Panax ginseng CA Meyer) belonging to the Araliaceae family as it is or by removing the thin root and cork layer. Its main pharmacological component is ginsenoside, which is a triterpenoid glycoside with a dammarene skeleton, which is only found in plants of the genus Panax ginseng.

In the present invention, the licorice (Glycyrrhizae Radix et Rhizoma) is a perennial plant belonging to the leguminous family (Leguminosae), which is the root of Glycyrrhiza uralensis Fischer, Glycyrrhiza glabra Linne, or Glycyrrhiza inflata Batal. and crude drugs using rhizomes as they are or those from which the perianth has been removed. Since ancient times, representative pharmacological actions of licorice have been known to include detoxification, choleretic phlegm, anti-ulcer, anti-inflammatory, antispasmodic, antitussive, and liver protection. In the present invention, one derived from licorice ( Glycyrrhiza uralensis Fischer) was used.

In the present invention, the peony (芍藥, Paeoniae Radix) is the root of the peony ( Paeonia lactiflora Pallas) belonging to the peony family (Paeoniaceae), and is also called white peony. Peony is known for its anti-inflammatory effect, analgesic effect, and digestive disorder improvement effect.

In the present invention, the at least one extract selected from the group consisting of ginseng, licorice and peony may be a single extract of ginseng, licorice and peony, or a mixed extract of ginseng, licorice and peony. In other words, it may be a ginseng extract, licorice extract, peony extract or a mixed extract of ginseng, licorice and peony. Preferably, it may be a mixed extract of ginseng, licorice and peony.

In the present invention, the mixed extract may be prepared by mixing single extracts obtained from each of ginseng, licorice and peony, and preferably may be prepared by mixing ginseng, licorice and peony and then extracting the mixture, but is limited thereto. It doesn't work.

In the present invention, the mixed extract may be extracted by mixing ginseng, licorice and peony at a weight ratio of 0.1 to 10:0.1 to 10:0.1 to 10. Preferably, it may be extracted by mixing ginseng, licorice, and peony at a weight ratio of 0.5 to 3:0.5 to 3:0.5 to 3, and when mixed at a weight ratio in the above range, it is similar to a single extract of ginseng, licorice, and peony. Or, it can exert a better prostatic hypertrophy inhibitory effect, and if it is out of the above range, the effect may be reduced or toxicity may appear than that of a single extract. More preferably, it may be extracted by mixing at a weight ratio of 1.1 to 3: 1: 1 or 1: 1.1 to 3: 1, and when mixed at a weight ratio in the above range, a synergistic effect appears in suppressing prostate hyperplasia, resulting in a remarkable effect can exert More preferably, it may be extracted by mixing at a weight ratio of 1.5 to 2.5: 1: 1 or 1: 1.5 to 2.5: 1, and most preferably, it may be extracted by mixing at a weight ratio of 1.5 to 2.5: 1: 1. , The weight ratio may be the most optimal mixing ratio condition to exert the effect of inhibiting prostate hypertrophy.

In one embodiment of the present invention, as a result of confirming the effect of inhibiting prostatic hypertrophy by varying the mixing ratio of ginseng, licorice and peony, ginseng: licorice: peony = mixed extract mixed at a ratio of 2: 1: 1 and 1: 2: 1 It was confirmed that the treatment showed an effect of inhibiting the expression of 5-alpha-reductase (5AR2), which was superior to that of the mixed extracts mixed at different ratios during treatment, and was effective in preventing and treating enlargement of the prostate by inhibiting it to a level similar to that of the normal group, In particular, it was confirmed that the mixed extract of ginseng: licorice: peony = 2: 1: 1 had the best effect (see Experimental Example 2).

In the present invention, the prostatic hyperplasia may be prostate caused by expression of 5 alpha-reductase, preferably prostate caused by expression of 5 alpha-reductase type 2, but is not limited thereto.

In the present invention, the extract may be one that inhibits the expression of 5 alpha-reductase to improve prostatic hyperplasia, and preferably inhibits the expression of 5 alpha-reductase type 2 to improve prostatic hyperplasia. Not limited.

Since the 5 alpha-reductase (5α-reductase) is the most important enzyme in the biosynthesis of dihydrotestosterone from testosterone, it is recognized as a major target for the control of benign prostatic hyperplasia. Among the three types of 5-alpha-reductase, type I (type 1) and type II (type 2) are clinically the most important. Among them, type 2, which is located in the stromal and epithelial nuclear membrane of the prostate, has been reported to be more involved in the production of dihydrotestosterone associated with benign prostatic hyperplasia. Testosterone and dihydrotestosterone play important roles in the development of the male reproductive system, but dihydrotestosterone has a higher affinity for the androgen receptor than testosterone. Dihydrotestosterone's high affinity for the androgen receptor can potently stimulate the transcriptional activity of androgen-dependent growth factors required to promote the proliferation of prostate epithelial and stromal cells rather than testosterone. This process requires the recruitment of specific androgen receptor coactivators to bind to DNA sequences of androgen response elements (AREs) that result from translocation of the receptor to the nucleus.

In one embodiment of the present invention, ginseng, licorice and peony extract alone and / or mixed extracts reduce the increased prostate weight in rats with prostatic hyperplasia, and the expression level of 5 alpha-reductase type 2 in human prostate epithelial cells. By reducing, it was confirmed that benign prostatic hyperplasia can be prevented, treated or improved (see Experimental Examples 1 to 4).

In the present invention, the extract may be one that does not reduce the sperm count of epididymis, and may have an effect of alleviating the side effects and toxicity of finasteride, an existing treatment for prostatic hyperplasia.

In one embodiment of the present invention, it was confirmed that the treatment of ginseng extract did not reduce the sperm count of epididymis of rats with prostatic hyperplasia, and when used in combination with finasteride, the decrease in sperm count, a side effect of finasteride, was improved. (See Experimental Example 5).

In the present invention, the 'extract' refers to a preparation obtained by squeezing an extraction target into an appropriate leachate or extraction solvent and evaporating the leachate or extraction solvent to concentrate, but is not limited thereto, an extract obtained by extraction, It may be a diluent or concentrate of an extract, a dried product obtained by drying the extract, and a crude or purified product thereof.

In the present invention, the extraction solvent may be water, alcohol or a mixture thereof. C1 to C2 lower alcohols may be used as the alcohol, and 30% ethanol, 50% ethanol, 70% ethanol or methanol may be used as the lower alcohol. Extraction methods may include, but are not limited to, vacuum extraction at high temperature, hot water extraction, reflux extraction, hot water extraction, cold needle extraction, room temperature extraction, ultrasonic extraction, or steam extraction. It may be extracted by adding the extraction solvent in an amount 1 to 20 times the volume (weight) of the extraction target. The extraction temperature may be 30 to 200 ° C, but is not limited thereto. In addition, the extraction time may be 1 to 24 hours, but is not limited thereto.

In the present invention, the vacuum concentration of the extract may be performed using a vacuum concentrator or a vacuum rotary evaporator, and the drying may be reduced pressure drying, vacuum drying, boiling drying, spray drying, or freeze drying.

In the present invention, 'prevention' means any action that suppresses or delays the progression of benign prostatic hyperplasia by administration of the composition of the present invention.

In the present invention, 'improvement' refers to all activities that improve or beneficially change symptoms of benign prostatic hyperplasia by administration of the composition of the present invention.

In the present invention, unless otherwise stated, 'treatment' refers to reversing, alleviating, inhibiting the progression of, or preventing the disease or disorder to which the term applies, or one or more symptoms of the disease or disorder. mean, and the term treatment as used herein refers to the act of treating when 'treating' is defined as above.

As used herein, the term 'administration' means providing the composition of the present invention in a predetermined pharmaceutically effective amount to a subject by any suitable method.

As used herein, the term 'pharmaceutically effective amount' means an amount sufficient to treat a disease with a reasonable benefit or risk ratio applicable to medical treatment, which is based on the type, severity, activity of a drug, drug It may be determined according to factors including sensitivity to, time of administration, route of administration and excretion rate, duration of treatment, drugs used concurrently, and other factors well known in the medical field.

The pharmaceutical composition of the present invention may be preferably formulated as a pharmaceutical composition by including one or more pharmaceutically acceptable carriers in addition to the active ingredient for administration. When formulated in the form of a liquid solution, it is sterile and biocompatible, and saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, or a mixture thereof can be used as a carrier. And, if necessary, other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added. Alternatively, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to prepare formulations for injections such as aqueous solutions, suspensions, emulsions, etc., pills, capsules, granules, or tablets.

In addition, the dosage form and pharmaceutically acceptable carrier of the pharmaceutical composition of the present invention are not particularly limited and may be appropriately selected according to known techniques in the art. In addition, an effective amount for the treatment or prevention of various immune-related diseases is the type of disease, the severity of the disease, the type and amount of active ingredients and other ingredients contained in the composition, the type of formulation and the patient's age, weight, general health condition, It can be adjusted according to various factors including gender and diet, administration time, route of administration and excretion rate of the composition, duration of treatment, and concurrently used drugs. The daily dosage is 0.0001 to 100 mg/kg, preferably 0.01 to 100 mg/kg, based on the amount of the mixed extract of the present invention, and may be administered 1 to 6 times a day. However, it is obvious to those skilled in the art that the dosage or dosage of each active ingredient should be such that the content of each active ingredient is not too high to cause side effects.

The pharmaceutical composition of the present invention can be administered to mammals such as rats, mice, livestock, and humans through various routes. All modes of administration can be envisaged, for example by oral, rectal or intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.

In addition, the present invention provides a food composition for preventing or improving prostatic hyperplasia comprising at least one extract selected from the group consisting of ginseng (Panax ginseng), licorice (Glycyrrhiza uralensis) and peony (Paeonia lactiflora) as an active ingredient. .

The term "food" means a natural product or processed product containing one or more nutrients, and preferably means a product that can be eaten directly through a certain degree of processing. , food additives, functional foods and beverages.

The food composition of the present invention can be formulated in the same way as the pharmaceutical composition and used as a functional food or added to various foods.

Foods to which the food composition can be added include, for example, various foods, beverages, chewing gum, tea, vitamin complexes, and functional foods. In addition, special nutritional foods (e.g., formula milk, infant food, baby food, etc.), processed meat products, fish meat products, tofu, jelly, noodles (e.g., ramen, noodles, etc.), breads, health supplements, seasoning foods (e.g., soy sauce) , soybean paste, gochujang, mixed paste, etc.), sauces, confectionery (e.g., snacks), candies, chocolates, chewing gum, ice cream, dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various types of kimchi) , pickles, etc.), beverages (eg, fruit drinks, vegetable drinks, soy milk, fermented beverages, etc.), natural seasonings (eg, ramen soup, etc.), but are not limited thereto. The food, beverage or food additive may be prepared by a conventional manufacturing method.

In addition, the above “functional food” or “health functional food” refers to a food group or food composition that has added value so that the food functions for a specific purpose by using physical, biochemical, or bioengineering methods. It refers to food designed and processed to fully express the body's regulatory functions related to defense rhythm control, disease prevention and recovery, etc., specifically, it may be a health functional food. The functional food may include food additives that are acceptable in food science, and may further include appropriate carriers, excipients, and diluents commonly used in the manufacture of functional foods.

The food composition of the present invention, in addition to containing an extract as an active ingredient, various nutrients, vitamins, minerals (electrolytes), flavors, flavors such as synthetic flavors and natural flavors, colorants and heavy substances like conventional food compositions. (cheese, chocolate, etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohol, carbonating agents used in carbonated beverages, natural carbohydrates, etc. may contain In addition, the food composition of the present invention may contain fruit flesh for preparing natural fruit juice, fruit juice beverages, and vegetable beverages.

Examples of the aforementioned natural carbohydrates include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrins and cyclodextrins, and sugar alcohols such as xylitol, sorbitol and erythritol. As the flavoring agents described above, natural flavoring agents (thaumatin), stevia extracts (eg rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can advantageously be used.

The food composition of the present invention may be provided as a health functional food or a health functional food composition, and may be prepared and processed in the form of tablets, capsules, powders, granules, liquids, pills, beverages, and the like.

The health functional food of the present invention may contain ordinary food additives, and the suitability as a food additive is determined according to the general rules of the Food Additive Code and General Test Methods approved by the Food and Drug Administration, unless otherwise specified. It is judged according to standards and standards. Examples of the items listed in the 'Food Additive Code' include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; natural additives such as persimmon pigment, licorice extract, crystalline cellulose, kaoliang pigment, and guar gum; and mixed preparations such as sodium L-glutamate preparations, noodle-added alkali preparations, preservative preparations, and tar color preparations. For example, a health functional food in the form of a tablet is obtained by granulating a mixture obtained by mixing the active ingredient of the present invention with an excipient, a binder, a disintegrant, and other additives in a conventional manner, and then adding a lubricant or the like to compression molding, or as described above. The mixture can be directly compression molded. In addition, the health functional food in the form of a tablet may contain a flavoring agent and the like as needed. Among health functional foods in the form of capsules, hard capsules can be prepared by filling a mixture in which the active ingredient of the present invention is mixed with additives such as excipients in a normal hard capsule. It can be prepared by filling the mixture mixed with gelatin in a capsule base. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary. The health functional food in the form of a pill can be prepared by molding a mixture of the active ingredient of the present invention mixed with an excipient, a binder, a disintegrant, etc. by a conventionally known method, and can be coated with sucrose or other coating agent if necessary, Alternatively, the surface may be coated with a material such as starch or talc. Health functional food in the form of granules can be prepared in granular form by a conventionally known method of mixing the active ingredient of the present invention with excipients, binders, disintegrants, etc., and, if necessary, flavoring agents, flavoring agents, etc. can

In addition, the present invention provides a method for preventing or treating prostatic hyperplasia comprising administering to a patient the pharmaceutical composition for preventing or treating BPH according to the present invention.

The treatment method of the present invention comprises administering to a subject a therapeutically effective amount of the pharmaceutical composition. A specific therapeutically effective amount for a specific individual depends on the type and degree of response to be achieved, the specific composition, including whether other agents are used as the case may be, the age, weight, general health condition, sex and diet of the individual, the time of administration, It is preferable to apply differently according to various factors including the route of administration and secretion rate of the composition, treatment period, drugs used together with or concurrently used with the specific composition, and similar factors well known in the medical field. Therefore, the effective amount of the composition suitable for the purpose of the present invention is preferably determined in consideration of the above.

The patient is applicable to any mammal, and the mammal includes domestic animals such as cattle, pigs, sheep, horses, dogs and cats as well as humans and primates.

Hereinafter, the present invention will be described in more detail through examples. These examples are intended to explain the present invention in more detail, and the scope of the present invention is not limited to these examples.

<Example 1> Preparation of ginseng, licorice and peony extracts

Using the following method, hot water extracts of ginseng, licorice and peony were prepared, and mixed extracts were prepared by varying their mixing ratio.

1-1. Preparation of ginseng extract

Ginseng ( Panax ginseng CA Mayer) 100g was put in 1L of distilled water, boiled at 100 ° C. for 150 minutes, and the extracted solution was freeze-dried to prepare a ginseng extract. The extraction yield was 15.23%.

1-2. Preparation of licorice extract

Licorice ( Glycyrrhiza uralensis Fiech.) 100 g was put in 1 L of distilled water, boiled at 100 ° C. for 150 minutes, and the extracted solution was freeze-dried to prepare a licorice extract. The extraction yield was 12.45%.

1-3. Preparation of peony extract

Peony ( Paeonia lactiflora ) 100g was put in 1L of distilled water, boiled at 100 ° C. for 150 minutes, and the extracted solution was freeze-dried to prepare a peony extract. The extraction yield was 10.12%.

1-4. Preparation of mixed extracts of ginseng, peony and licorice

Ginseng, licorice and peony of Examples 1-1 to 1-3 were mixed in a weight ratio of 1: 1: 1, 2: 1: 1, 1: 2: 1 and 1: 1: 2, and then the mixture 100 g was put into 1L of distilled water, boiled at 100℃ for 150 minutes, and the extracted solution was lyophilized to obtain a mixed extract of ginseng, peony and licorice with different mixing ratios (ginseng: licorice: peony = 1:1:1, 2:1 :1, 1:2:1 and 1:1:2) were prepared.

<Experimental Example 1> Confirmation of cytotoxicity of the extract

In order to confirm the cytotoxicity of ginseng, licorice and peony extracts alone and mixed extracts, cell viability was measured using RWPE-1 cells, which are human prostate epithelial cells.

The cells were cultured in each well at 1 x 10 ⁵ cells/200μl in a 96-well plate, and CO ₂ It was cultured for 24 hours in a 36° C. cell incubator maintained at 5%. Each extract prepared in Example 1 was diluted to 10 mg/ml using DMSO as a solvent and treated in each well at the concentration shown in FIG. 1. The plate treated with the drug is CO ₂ It was cultured for 24 hours in a 36° C. cell incubator maintained at 5%. Then, after replacing with a new culture medium, 10 μl/200 μl of WST-1 cell proliferation assay mixture (Biomax, Seoul, Korea) was added to each well to ensure that CO ₂ It was cultured for 4 hours in a 36° C. cell incubator maintained at 5%. The cultured plate was measured for absorbance at a wavelength of 440 nm using a spectrophotometer.

As shown in Figure 1, as a result of quantifying the cytotoxicity based on the well containing DMSO alone, ginseng extract, peony extract, ginseng: licorice: peony = 1: 1: 1 mixed extract, ginseng: licorice: peony = 2 :1:1 mixed extract and ginseng:licorice:peony = 1:2:1 mixed extract were 1000μg/ml, and cytotoxic up to a concentration of 500μg/ml in licorice extract and ginseng:licorice:peony = 1:1:2 mixed extract It was confirmed that there was no (Fig. 1). Therefore, in Experimental Example 2, each extract was treated at a concentration of 500 μg/ml and the experiment was conducted.

<Experimental Example 2> Measurement of 5 alpha-reductase inhibition effect according to extract treatment

8 x 10 RWPE-1, human prostate epithelial cells⁵/2ml incubated with CO₂go It was cultured for 24 hours in a 36° C. cell incubator maintained at 5%. After replacing the culture medium on the plate with a new one, 4 μM of testosterone propionate (TP; Wako pure chemical industries, Osaka, Japan) was pretreated for 30 minutes. Thereafter, the mixed extract of ginseng, licorice and peony at different mixing ratios of Example 1 was treated at a non-toxic concentration (500 μg/ml) measured in Experimental Example 2, and CO₂go It was cultured for 12 hours in a 36° C. cell incubator maintained at 5%. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed on cell models that induced prostatic hyperplasia, and 5-alpha-reductase type 2 (5α-reductase-2, 5AR2) ) The effect of inhibiting gene (mRNA) expression was analyzed.

RNA was extracted from cells treated with mixed extracts by mixing ratio using QIAzol lysis reagent. RNA obtained from each experimental group was prepared as single stranded cDNA using a power cDNA synthesis kit (iNtRON Biotechnology, Seongnam, Kyunggi, Korea). 5AR2 primers that specifically amplify DNA were put in and the mRNA expression level was measured using real-time RTPCR (Applied Biosystems, Foster City, CA, USA), and the expression level was quantified based on the normal group (B) The results are shown in Figure 2.

As shown in Figure 2, in the case of the disease control group ( TP group ), the expression of 5 alpha-reductase type 2 (5AR2) mRNA was significantly increased compared to the normal group ( Group B ), but ginseng, licorice and peony mixed extracts treated In the case of the experimental group, it was confirmed that the increase in the expression of 5-alpha-reductase was alleviated. In particular, expression was significantly suppressed in the groups treated with the ginseng: licorice: peony = 2:1:1 and 1:2:1 mixed extracts, and the ginseng: licorice: peony = 2:1:1 mixed extract treated group ( 2 :1:1 group ), it was confirmed that 5AR2 expression was suppressed at a level almost similar to that of the normal group ( group B ).

Therefore, the composition comprising the extracts of ginseng, licorice and peony of the present invention as active ingredients can be effectively used for the prevention, treatment and improvement of benign prostatic hyperplasia, and ginseng: licorice: peony = 2: 1: 1 and 1: 2: It was confirmed that the mixed extract mixed at a weight ratio of 1, in particular, the mixed extract of ginseng: licorice: peony = 2: 1: 1 is the optimal condition for exhibiting the most excellent prostatic hyperplasia prevention, treatment and improvement effects.

<Experimental Example 3> Preparation of experimental animals with prostatic hyperplasia induced

A 12-week-old male Sprague-Dawley (SD) rat (200 ± 20 g) was purchased from Daehan Biolink (Eumseong, Korea) for an animal model of benign prostatic hyperplasia and was tested. After a 1-week adaptation period before the start of the experiment, food and water were freely consumed for 6 weeks during the experiment. . In addition, testosterone propionate (TP) for inducing prostatic hyperplasia was purchased from Wako pure chemical industries (Osaka, Japan), and finasteride (Fi) (≥ 97% pure) for use as a positive control was Sigma-Aldrich Inc. (MO, USA) was purchased and used. Each extract powder obtained by lyophilization in Example 1 was dissolved in tertiary distilled water, filtered through a 22 μm filter, and used in the experiment.

Rats were divided into 8 groups to induce prostatic hyperplasia, and 6 animals were randomly assigned to each group. Among them, 7 groups were injected with testosterone propionate (TP; 5 mg/kg/day) into the groin for 4 weeks, and the other group, a normal control group, was injected with the same amount of ethanol. After prostatic hyperplasia induction for 4 weeks, (i) a normal control group injected with ethanol for an additional 4 weeks ( NC group ), (ii) a disease control group injected with TP for an additional 4 weeks (benign prostatic hyperplasia, BPH group ), ( iii) Experimental groups injected with ginseng extract (20 mg/kg/day), licorice extract (40 mg/kg/day), or peony extract (80 mg/kg/day) together with TP for an additional 4 weeks ( Ginseng group , licorice group) group and peony group ), (iv) a positive control group injected with finasteride (Fi; 1 mg/kg/day), a treatment for prostatic hyperplasia, along with TP for an additional 4 weeks ( Fi group ), and (v) TP for an additional 4 weeks Combined treatment group treated with finasteride (1 mg/ kg /day) and ginseng extract (20 mg/kg/day) or licorice (40 mg/kg/day) together +Fi group ), the experiment was conducted with different treatments.

<Experimental Example 4> Prostate tissue hypertrophy inhibitory effect by the mixed extract

The present inventors conducted an experiment to confirm whether the extracts of ginseng, peony and licorice had an effect of inhibiting prostate tissue hypertrophy.

In order to confirm the induction of benign prostatic hyperplasia and improvement effects of ginseng, peony and licorice extracts, the volume and weight of prostate tissue of rats were measured after a total of 8 weeks of experimentation. In addition, in order to compensate for the difference in tissue weight according to body weight, the prostate index was calculated by dividing the weight mg of the prostate with respect to 100 g of body weight, and is shown in FIGS. 3 to 5.

As a result of measuring the weight of prostate tissue, it was confirmed that the total prostate weight increased significantly by more than 200% in the BPH group compared to the NC group. As a result of the prostate index correcting the weight of prostate tissue for body weight, it was confirmed that the disease control group ( BPH group ) increased significantly by more than 200% compared to the normal control group ( NC group ). In the experimental groups treated with the mixed extract ( ginseng group , licorice group, and peony group ), enlargement of prostate tissue was significantly suppressed compared to the disease control group ( BPH group ) in which prostatic hyperplasia was induced, and the positive control group injected with finasteride ( Fi group ) It was confirmed to suppress prostate hypertrophy at a level similar to that reduced compared to the BPH group (FIGS. 3 to 5). In addition, in the case of the combined treatment group (licorice + Fi) treated with finasteride and licorice, prostatic hypertrophy was significantly suppressed compared to the case of licorice or finasteride alone injection, and the enlargement of the prostate was similar to that of the normal control group (NC group). It was confirmed that it decreased (FIG. 4).

Therefore, it was confirmed that the ginseng, licorice and peony extracts and the mixed extracts obtained by mixing them were effective in preventing or treating benign prostatic hyperplasia.

<Experimental Example 5> Effects of mitigating the side effects of the extract on the side effects of finasteride, a conventional treatment

Finasteride, previously used, was effective for benign prostatic hyperplasia, but had a serious side effect of reducing sperm count. Accordingly, the present inventors confirmed whether ginseng, licorice, and peony extracts had an effect on sperm count.

In order to check the sperm count, the cauda epididymis of the SD rats used in Experimental Examples 4 and 5 was excised and cut into small pieces twice. Thereafter, the cells were soaked in 4 ml of PBS for 1 hour to allow sperm to spread into the PBS solution. PBS containing sperm was mixed with an equal amount of methanol and observed using a hematocytometer and an Olympus IX71 Research Inverted Phase microscope (Olympus Co., Tokyo, Japan).

As a result of confirming the change in sperm count in the epididymis, the sperm count in the finasteride-treated group ( Fi group ) was significantly reduced compared to the disease control group ( BPH group ), whereas the sperm count in the experimental group ( Gi group ) treated with ginseng extract was significantly different. However, in the group administered finasteride and ginseng together ( Gi+Fi group ), the decrease in sperm count, a side effect of finasteride, was improved (FIG. 6).

Therefore, it was confirmed that the ginseng extract of the present invention and the mixed extract of ginseng, peony, and licorice containing the same showed an effect on benign prostatic hyperplasia without reducing the sperm count, compared to finasteride, a conventional treatment for prostatic hyperplasia.

So far, the present invention has been looked at mainly with its preferred embodiments. Those skilled in the art to which the present invention pertains will be able to understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments should be considered from a descriptive point of view rather than a limiting point of view. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the equivalent scope will be construed as being included in the present invention.

Claims (11)

A pharmaceutical composition for preventing or treating benign prostatic hyperplasia comprising a mixture of ginseng (Panax ginseng), licorice (Glycyrrhiza uralensis) and peony (Paeonia lactiflora) as an active ingredient,
The mixture was extracted with hot water,
The mixture inhibits the expression of 5 alpha-reductase,
The mixture is extracted by mixing ginseng, licorice and peony in a weight ratio of 2: 1: 1, the composition. delete delete delete delete delete delete A food composition for preventing or improving prostatic hyperplasia comprising a mixture of ginseng (Panax ginseng), licorice (Glycyrrhiza uralensis) and peony (Paeonia lactiflora) as an active ingredient,
The mixture was extracted with hot water,
The mixture inhibits the expression of 5 alpha-reductase,
The mixture is extracted by mixing ginseng, licorice and peony in a weight ratio of 2: 1: 1, the composition. delete delete A method for preventing or treating enlargement of the prostate comprising administering the pharmaceutical composition for preventing or treating enlargement of the prostate of claim 1 to a non-human patient.

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