CN112638394A - Composition for preventing or treating prostate disease comprising Japanese stauntonvine extract and dogwood extract as active ingredients - Google Patents
Composition for preventing or treating prostate disease comprising Japanese stauntonvine extract and dogwood extract as active ingredients Download PDFInfo
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- CN112638394A CN112638394A CN201980055052.XA CN201980055052A CN112638394A CN 112638394 A CN112638394 A CN 112638394A CN 201980055052 A CN201980055052 A CN 201980055052A CN 112638394 A CN112638394 A CN 112638394A
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Abstract
The present invention relates to a composition for preventing or treating prostate diseases comprising extracts of chaenomeles japonicas and cornus officinalis as active ingredients, and more particularly, to a composition for preventing or treating prostate diseases, which comprises a compound extract of chaenomeles japonicas and cornus officinalis as an active ingredient, has no cytotoxicity, and has an excellent inhibitory effect on the expression of androgen receptors, prostate-specific antigens and prostate-specific enzymes, and thus can be used as an active ingredient of a composition for preventing or treating prostate diseases.
Description
Technical Field
The present invention relates to a composition for preventing or treating prostate diseases, which comprises an extract of Stauntonia hexaphylla and an extract of Cornus officinalis (Cornus officinalis sieb. et Zucc) as active ingredients.
Background
The prostate gland is an accessory gonad composed of glandular tissue and fibromuscular tissue, and plays a role in producing and secreting semen. The prostatic fluid produced in the prostate gland supplies nutrients to the sperm that are produced and move in the testis and helps the sperm to move actively by maintaining the fluid state so that the ejaculated semen does not coagulate. Typical prostate diseases include prostatic hyperplasia, prostate cancer and prostatitis.
Prostatic Hyperplasia (BPH) is one of the diseases commonly seen in adult men, and it is reported that the incidence rate of men over 50 years old is 50%, and that the incidence rate of men over eighty years old is 80% or more, and the frequency of occurrence in male urinary tract disorders is the highest. The number of patients with prostatic hyperplasia in korea has increased from 45 ten thousand to 9 thousand in 2006 to 84 ten thousand to 2 thousand in 2011, and has a growth rate of 83.5%, and recently, the incidence of the disease has sharply increased.
Generally, the size of the prostate of an adult is about 20g, and if the prostate is enlarged to an abnormal size and reaches 40 to 400g, the prostate presses the nearby urinary tract, and therefore, there are symptoms such as urinary bladder urine storage symptoms such as frequent urination 8 times or more a day, frequent urination at night, urinary urge in which urination is not suppressed because a strong and sudden urinary sensation is felt, urinary waiting for urination until urination is achieved by a urinary delay or a urinary interruption, and urinary bladder discharge disorders such as a need to apply force during urination. Prostate hyperplasia does not require surgical treatment without urinary occlusion or other complications, but about 50% of patients by age 80 still require treatment. Moreover, some of patients with prostatic hyperplasia are related to prostate cancer, and although this is not a fatal disease, it is becoming an important medical problem in the current society that is advancing to the super-aging society in terms of reducing the quality of life.
At the beginning, the treatment of prostatic hyperplasia was almost surgical, but the method of administration was rapidly changing from the second half of the last eighties. However, since the conventional drugs for treating prostatic hyperplasia have many side effects and limited efficacy, development of a novel therapeutic agent having no side effects, no drug resistance, and excellent therapeutic effect is required.
Prostate cancer is the most common cancer among male cancers in the west, and presents a very high incidence rate, which is also rapidly increasing in korea. The treatment of prostate cancer varies depending on the stage of disease, and in the case of local cancer, treatment is performed for radical treatment, but in the case of metastatic cancer, systemic treatment is required. The local prostate cancer may be selected from environmental therapy, radical prostatectomy, and radiation therapy in consideration of the age, health status, sexual function status, the change stage and differentiation degree of tumor, preference degree of patient, etc. Metastatic prostate cancer can be treated with hormone and anticancer therapy.
Prostatitis, a disease in which the prostate gland produces inflammation, is a very common disease in which about 50% of adult men experience the symptoms once in a lifetime. Urinary infection is often caused by direct infection of bacteria through the urinary tract, and may also be caused by disturbance of excretion of prostatic fluid, backflow of urine into the prostate, infection with inflammation such as hemorrhoid and colitis, and the like.
Since the above prostate diseases such as prostate cancer, prostatic hyperplasia and prostatitis are related to androgen which is a male hormone, therapeutic and prophylactic effects on diseases or symptomatic relief effects can be obtained by anti-androgen drugs which inhibit the expression of androgen receptors. In addition, a prostate-specific antigen inhibitor that inhibits the expression of a prostate-specific antigen is also effective for the prevention and treatment/symptom improvement of the above-mentioned prostate diseases that are affected by the prostate-specific antigen. Meanwhile, 5-alpha reductase inhibitors which inhibit androgen are also used for the treatment of male diseases caused by androgen, such as prostatic hyperplasia, alopecia, etc. More specifically, 5- α reductase, which converts testosterone (testosterone), which is a male hormone, into Dihydrotestosterone (DHT), is used as reduced coenzyme ii (nadph) -dependent 3-oxo 5 α -steroid dehydrogenase (3-oxo-5 α -steroid-dehydrogenase) of microsomes (microsomes) to reduce doubly bound steroids such as testosterone. The 5-alpha reductase has subtypes of type 1, type 2, and type 3, with type 1 being mainly present in several organs such as skin and liver tissue, and type 2 being mainly present in liver and genitourinary including prostate. In prostate tissue, more than 90% of testosterone is converted to dihydrotestosterone by 5-alpha reductase, and dihydrotestosterone promotes prostate growth with about 10 times more activity than testosterone. Therefore, if 5-alpha reductase is effectively inhibited, the synthesis of dihydrotestosterone, which has strong physiological activity, can be reduced, and finally prostatic hyperplasia can be improved or even treated. In addition, in the same way as in prostatic hyperplasia, when dihydrotestosterone is synthesized excessively by 5-alpha reductase, dihydrotestosterone binds to androgen receptors around hair follicles, gradually shrinking and degenerating the hair follicles, and thus causing alopecia. Thus, alopecia can be improved or even treated by effectively inhibiting 5-alpha reductase.
On the other hand, Stauntonia chinensis, which is a plant of evergreen vine of the family akebiaceae and is known as Stauntonia Hexaphylla, is found in streams or bushes in southern regions of korea, such as the whole south, kyo south, and loyalty south. The stem, leaf and root are used as medicinal materials and are known to have significant effects on resisting inflammation, relieving fever and pain. And may also be used as a cardiotonic agent to restore function to normal when the heart is weakened or incomplete, or as a diuretic agent to expel excess water from the body. In addition, it is known that it is effective for inhibiting subcutaneous fat accumulation, joint diseases, uterine fibroids, facial neuralgia, and the like.
Cornus officinalis (Corni Fructus) is a ripe, depeded pulp derived from Cornus officinalis Siebold et Zuccarini (Cornaceae). The bag-shaped plastic bag is irregular block-shaped or bag-shaped, and has a length of about 10-15 mm and a width of about 1 cm. The outer surface is dark red to dark purple, and the surface has moist and rough wrinkles. The pulp has the trace of removing seeds, the upper part has the trace of calyx, and the lower part has the trace of fruit stem. It has soft meat quality, slight smell, and sour and sweet taste. It is known that cornus officinalis has diuretic, antihypertensive, tonic, antiallergic, antihistaminic, anticholinergic, antipbarium, antimicrobial (antifungal) and other effects.
Accordingly, the present inventors have made an effort to develop a therapeutic agent for prostate diseases derived from natural products, which ensures safety and has no side effects, and have confirmed that a complex extract of chaenomeles speciosa and cornus officinalis has no cytotoxicity and an excellent inhibitory effect on the expression of androgen receptors, prostate-specific antigens and prostate-specific enzymes, and have found that the compound extract can be used as an active ingredient of a composition for preventing or treating prostate diseases, thereby completing the present invention.
Disclosure of Invention
Technical problem to be solved by the invention
The present invention is directed to a pharmaceutical composition for preventing or treating prostate diseases, comprising extracts of Japanese stauntonvine and Cornus officinalis as active ingredients.
It is still another object of the present invention to provide a method for preventing or treating a prostate disease, which comprises the step of administering pharmaceutically effective amounts of the extracts of chaenomeles japonicas and cornus officinalis to an individual.
Another object of the present invention is to provide a use of the japanese stauntonvine extract and the dogwood extract as a composition for preventing or treating prostate diseases.
Means for solving the problems
In order to achieve the above objects, the present invention provides a pharmaceutical composition for preventing or treating prostate diseases, comprising extracts of chaenomeles japonicas and cornus officinalis as effective ingredients.
Also, the present invention provides a health food for preventing or improving prostate diseases, which comprises the extract of chaenomeles speciosa and the extract of cornus officinalis as effective ingredients.
Also, the present invention provides a food composition for preventing or improving prostate diseases, comprising the extract of chaenomeles speciosa and the extract of cornus officinalis as active ingredients.
Also, the present invention provides a method for treating prostate diseases, which comprises the step of administering pharmaceutically effective amounts of the extracts of chaenomeles japonicas and cornus officinalis to an individual.
Also, the present invention provides a method for preventing or ameliorating a prostate disease, comprising the step of administering a pharmaceutically effective amount of the extracts of Japanese stauntonvine and Corni fructus to an individual.
Also, the present invention provides a use of the Japanese stauntonvine extract and the Cornus officinalis extract as a pharmaceutical composition for preventing or treating prostate diseases.
Also, the present invention provides a use of the Japanese stauntonvine extract and the Cornus officinalis extract as a health food for preventing or improving prostate diseases.
Meanwhile, the present invention provides a use of the Japanese stauntonvine extract and the dogwood extract as a food composition for preventing or improving prostate diseases.
ADVANTAGEOUS EFFECTS OF INVENTION
The compound extract of chaenomeles speciosa and dogwood of the present invention has no cytotoxicity, has excellent inhibitory effects on the expression of androgen receptors, prostate specific antigens and prostate specific enzymes, and can be used as an active ingredient of a composition for preventing or treating prostate diseases.
Drawings
FIG. 1 is a graph showing the confirmation of cytotoxicity after treating human prostate cancer (LNCaP) cells with the Japanese stauntonia extract, the Corni fructus extract or the Japanese stauntonia and Corni fructus complex extract prepared in the examples of the present invention at concentrations of 0. mu.g/ml, 10. mu.g/ml, 25. mu.g/ml and 50. mu.g/ml, respectively.
FIG. 2 is a graph showing the confirmation of the change in the expression of Androgen Receptor (AR), Prostate Specific Antigen (PSA) and 5-alpha reductase (5 alpha-reductase) 2 proteins after treating human prostate cancer cells with 1. mu.M testosterone in combination with 25. mu.g/ml, 50. mu.g/ml concentration of Japanese stauntonvine extract, 50. mu.g/ml concentration of Corni fructus extract or 25. mu.g/ml concentration of Japanese stauntonvine and Corni fructus complex extract prepared in the present examples.
Detailed Description
The present invention will be described in more detail below.
The present invention provides a pharmaceutical composition for preventing or treating prostate diseases comprising Japanese stauntonvine extract and dogwood extract as active ingredients.
Also, the present invention provides a use of the Japanese stauntonvine extract and the Cornus officinalis extract as a pharmaceutical composition for preventing or treating prostate diseases.
Preferably, the japanese stauntonvine extract and the dogwood extract, which are the effective ingredients of the present invention, are prepared by a method comprising the following steps, respectively, but not limited thereto:
step 1), adding an extraction solvent to the Japanese stauntonvine and the dogwood respectively for extraction;
step 2), filtering the extract of step 1);
step 3), concentrating the filtrate obtained in the step 2) under reduced pressure;
and step 4) drying the concentrate of step 3).
In the preparation method of the present invention, the Japanese stauntonvine or the dogwood of the step 1) can be used without limitation, whether cultivated or marketed. Also, the above Japanese stauntonia may be flower, branch, stem, leaf, fruit, aerial part, rhizome, root or combination thereof of Japanese stauntonia, more specifically, Japanese stauntonia leaf.
In the preparation method of the present invention, preferably, the extraction solvent of the above step 1) may use water, alcohol or a mixture thereof. Preferably, the above alcohol is used C1To C2Preferably, a lower alcohol ofEthanol or methanol is used. Preferably, the extraction method uses shaking extraction, Soxhelt extraction, or reflux extraction, but is not limited thereto. Preferably, the extraction solvent is added in an amount of 1 to 20 times, more preferably 3 to 17 times, and still more preferably 5 to 15 times the amount of the dried Japanese stauntonvine or dogwood. Preferably, the extraction temperature is 10 to 100, but is not limited thereto. And, preferably, the extraction time is 1 to 72 hours, but is not limited thereto. Meanwhile, the number of extraction is preferably 1 to 5, more preferably 3 to 4, and further preferably 3, but not limited thereto.
In the preparation method of the present invention, preferably, the reduced pressure concentration of step 3) uses a vacuum reduced pressure concentrator or a vacuum rotary evaporator, but is not limited thereto. Also, preferably, the drying is reduced pressure drying, vacuum drying, boiling drying, spray drying or freeze drying, but is not limited thereto.
In the preparation method of the present invention, the extract of japanese stauntonvine and the extract of dogwood may be prepared by mixing the respective extracts after preparing the extracts, or may be prepared by mixing japanese stauntonvine and dogwood and obtaining their extracts after mixing them.
The above composition may contain the extract of Stauntonia japonica and the extract of Cornus officinalis at a weight ratio of 5: 5 to 9.5: 0.5, specifically 6: 4 to 9.5: 0.5, more specifically 7: 3 to 9.5: 0.5, further specifically 8: 2 to 9.5: 0.5, further specifically 8.5: 2.5 to 9.5: 0.5.
The prostate disease may be, for example, prostate cancer, prostatic hyperplasia or prostatitis.
In an embodiment of the present invention, the present inventors prepared a compound extract of japanese stauntonia chinensis and dogwood by mixing the above extracts in various mixing ratios after preparing a 70% alcohol extract of japanese stauntonia chinensis leaves and a 70% alcohol extract of dogwood.
The present inventors also confirmed the cytotoxicity of the above-mentioned compound extract of chaenomeles speciosa and dogwood in prostate cancer cells, and confirmed that the compound extract was not cytotoxic at various concentrations (see fig. 1).
The present inventors also confirmed that the complex extract has an excellent effect of inhibiting the expression of a protein of Androgen Receptor (AR), Prostate Specific Antigen (PSA), and prostate specific enzyme (5- α reductase 2) in prostate cancer cells treated with testosterone together with the complex extract of chaenomeles speciosa and dogwood (see fig. 2).
Accordingly, the present inventors confirmed that the complex extract of chaenomeles speciosa and dogwood of the present invention has no cytotoxicity and excellent inhibitory effects on the expression of androgen receptors, prostate-specific antigens and prostate-specific enzymes, and thus the complex extract of chaenomeles speciosa and dogwood of the present invention can be effectively used as an active ingredient of a pharmaceutical composition for preventing or treating prostate diseases or a pharmaceutical composition for preventing or ameliorating male diseases caused by testosterone, such as alopecia.
The compositions of the invention may be administered orally or non-orally (e.g. by spreading or intravenous, subcutaneous, intraperitoneal injection). The preparations for non-oral administration may be used in the form of external preparations such as powder, granule, tablet, capsule, sterile aqueous solution, liquid, non-aqueous solvent, suspension, emulsion, syrup, suppository, aerosol, etc. and sterile injection, respectively, according to the usual methods, and preferably, may be used in the form of pharmaceutical compositions for external use for skin prepared as cream, gel, patch, spray, ointment, plaster, emulsion, liniment, paste or gel ointment, but are not limited thereto. The compositions for topical administration may be anhydrous or aqueous, depending on the clinical prescription. As the nonaqueous solvent or suspending agent, propylene glycol (propylene glycol), polyethylene glycol, vegetable oil such as olive oil, or injectable ester such as ethyl oleate can be used. The base of suppository can be semisynthetic fatty acid ester (witepsol), polyethylene glycol, tween (tween)61, cacao butter, hydroxycitronellal, glycerogelatin, etc. Solid preparations for oral administration include powders, granules, tablets, capsules, soft capsules, pills and the like. Liquid preparations for oral administration include suspensions, internal liquids, oils, syrups, aerosols and the like, and may contain excipients such as wetting agents, sweeteners, aromatics, preservatives and the like in addition to water and liquid paraffin, which are widely used as simple diluents.
For administration, the composition may be prepared by including one or more pharmaceutically acceptable carriers in addition to the active ingredient of the present invention. The pharmaceutically acceptable carrier may be physiological saline, sterilized water, ringer's solution, buffered physiological saline, glucose solution, maltodextrin solution, glycerol, ethanol, or a mixture thereof, and may further contain other common additives such as antioxidant, buffer, bacteriostatic agent, etc., as required. In addition, a diluent, a dispersant, a surfactant, a binder and a lubricant may be added to prepare an injection preparation such as an aqueous solution, a suspension, an emulsion and the like.
Preferably, the pharmaceutically acceptable additive of the present invention may be contained in an amount of 0.1 parts by weight to 90 parts by weight, relative to the above composition.
The preferred administration amount of the composition of the present invention varies depending on the degree of absorption of the active ingredient in the body, the age, sex and degree of obesity of the patient, and can be appropriately selected by one of ordinary skill in the art to which the present invention pertains. However, for preferable effects, in the case of oral administration dosage forms, it can be generally administered to an adult in an amount of 0.0001mg/kg to 500mg/kg per day, preferably 0.001mg/kg to 300mg/kg, more preferably 0.01mg/kg to 200 mg/kg. The administration can be carried out once a day or several times. The above dosage does not limit the scope of the present invention in any way.
Also, the present invention provides a health food for preventing or improving prostate diseases, which comprises the extract of chaenomeles speciosa and the extract of cornus officinalis as effective ingredients.
Also, the present invention provides a food composition for preventing or improving prostate diseases, comprising the extract of chaenomeles speciosa and the extract of cornus officinalis as active ingredients.
Also, the present invention provides a use of the Japanese stauntonvine extract and the Cornus officinalis extract as a health food for preventing or improving prostate diseases.
Also, the present invention provides a use of the Japanese stauntonvine extract and the Cornus officinalis extract as a food composition for preventing or improving prostate diseases.
The above-mentioned japanese stauntonvine extract and dogwood extract, their mixing ratio, and prostate disease are the same as those described above with respect to the pharmaceutical composition for preventing or treating prostate disease, and therefore the specific description refers to the above-mentioned contents, and only the specific structure of the health food or food composition will be described below.
On the other hand, it was confirmed that the complex extract of chaenomeles speciosa and dogwood of the present invention has no cytotoxicity and is excellent in the inhibitory effect on the expression of androgen receptor, prostate specific antigen and prostate specific enzyme, and thus the complex extract of the present invention can be effectively used as an active ingredient of health food or food composition for preventing or improving prostate disease. Also, the complex extract of the present invention can be effectively used as an active ingredient of a health food or food composition for preventing or improving male diseases caused by testosterone, such as alopecia.
The two extracts of the present invention may be provided as a health food or food composition in admixture with a dietetically acceptable carrier.
The health food of the invention comprises the forms of tablets, capsules, pills or liquid, etc.
When the two extracts of the present invention are used as additives for foods or drinks, the two extracts may be added as they are, or may be used together with other foods or food ingredients, and may be used as appropriate according to a common method. The mixing amount of the above two extracts may be appropriately determined depending on the purpose of use thereof (preventive, health or therapeutic treatment). In the case of long-term ingestion for the purpose of health and hygiene or for the purpose of health regulation, since both of the above extracts have no problem in terms of safety, they can be taken for a long period of time. The kind of the above-mentioned food is not particularly limited.
Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, sugar, snack, biscuit, pizza, stretched noodles, other noodles, chewing gum, milk products including ice cream, various soups, beverages, tea, drinkable preparation, alcoholic beverage, and vitamin complex. In the case of preparing a beverage, liquid components to be added in addition to the above two extracts are not limited, but a plurality of flavors, natural carbohydrates, and the like may be added as additional components as in the case of a normal beverage. Examples of the natural carbohydrates include monosaccharides (e.g., glucose, fructose, etc.), disaccharides (e.g., maltose, sucrose, etc.), polysaccharides (e.g., general sugars such as dextrin, cyclodextrin, etc.), and sugar alcohols such as xylitol, sorbitol, erythritol. The proportion of natural carbohydrates is between 1g and 20g, preferably between 5g and 12g, per 100ml of the composition according to the invention. In addition to the above, natural flavors (thaumatin, stevia extracts (e.g., rebaudioside a, glycyrrhizic acid, etc.)) and synthetic flavors (e.g., saccharin, aspartame, etc.) can be used.
The food composition of the present invention may contain various nutrients, vitamins, flavors such as inorganic substances (electrolytes), synthetic flavors and natural flavors, colorants and enhancers (cheese, chocolate and the like), pectic acid and salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohol, carbonating agents for carbonated beverages, and the like. Also, the food composition of the present invention may comprise pulp for preparing fruit and vegetable beverages. These ingredients may be used alone or in combination, and the proportion of these additives is generally selected in the range of 0.001 to 50 parts by weight relative to the total weight of the composition.
Also, the present invention provides a method for preventing or ameliorating a prostate disease, comprising the step of administering a pharmaceutically effective amount of the extracts of Japanese stauntonvine and Corni fructus to an individual.
Also, the present invention provides a method for treating prostate diseases, which comprises the step of administering pharmaceutically effective amounts of the extracts of Japanese stauntonvine and Corni fructus to an individual.
The above-mentioned Japanese stauntonvine extract and the extract of Cornus officinalis, the mixing ratio thereof, and the prostate disease are the same as those described above with respect to the pharmaceutical composition for preventing or treating prostate disease, and the specific description is incorporated above.
On the other hand, it was confirmed that the complex extract of chaenomeles speciosa and dogwood of the present invention has no cytotoxicity and an excellent inhibitory effect on the expression of androgen receptors, prostate specific antigens and prostate specific enzymes, and thus the complex extract of the present invention can be effectively used for the prevention, amelioration or treatment of prostate diseases. Also, the complex extract of the present invention can be effectively used for the prevention or improvement of male diseases caused by testosterone, such as alopecia.
The present invention will be described in detail below with reference to examples, experimental examples, and preparation examples.
However, the following examples, experimental examples and preparation examples merely illustrate the present invention, and the contents of the present invention are not limited to the following examples, experimental examples and preparation examples.
Example 1 preparation of Japanese Stauntonia extract
20g of dried Japanese stauntonia leaves were put in 300ml of 70% ethanol and subjected to low-temperature extraction by ultrasonic waves for 12 hours. After filtering the obtained extract using filter paper, the filtered residue was subjected to low-temperature extraction using 300ml of 70% alcohol for 12 hours and the filtering process was repeated two more times. After the extraction is finished, the filtrates are combined and the Japanese stauntonvine extract is obtained through the processes of decompression concentration and freeze drying.
Example 2 preparation of extract of Cornus officinalis
20g of dried Corni fructus was put in 300ml of 70% ethanol, and extracted at low temperature by ultrasonic wave for 12 hours. After filtering the obtained extract using filter paper, the filtered residue was subjected to low-temperature extraction using 300ml of 70% alcohol for 12 hours and the filtering process was repeated two more times. After extraction, combining the filtrates, and concentrating under reduced pressure and freeze drying to obtain Corni fructus extract.
Example 3 preparation of Compound extract of Japanese Stauntonia chinensis and Cornus officinalis
The compound extracts of japanese stauntonvine and dogwood were prepared by mixing the extract of japanese stauntonvine obtained in the above example 1 and the extract of dogwood obtained in the above example 2 at the mixing ratios shown in the following table 1.
TABLE 1
Experimental example 1 confirmation of cytotoxicity of Compound extract of Stauntonia japonica and Cornus officinalis
In order to observe the cytotoxicity of the compound extract of stauntonia japonica and dogwood, the cell survival rate was confirmed after prostate cancer cells were treated with the compound extract of stauntonia japonica and dogwood.
Specifically, LNCaP cells (ATCC company, USA) as a male hormone-dependent human prostate cancer cell line were cultured in RPMI-1640 medium (Gibco company) containing 10% Fetal Bovine Serum (FBS) at 37 ℃ and 5% CO2Culturing under the conditions of (1). The cultured cells were cultured in a 96-well plate at 1X 104The concentration of cells (cells)/well (well) was measured, and the cells were cultured under the same conditions as described above for 18 hours. Then, the Japanese stauntonvine extract prepared in the above example 1, the Corni fructus extract prepared in the above example 2, or the Japanese stauntonvine and Corni fructus complex extract prepared in the above example 3 were treated with concentrations of 0. mu.g/ml, 10. mu.g/ml, 25. mu.g/ml, and 50. mu.g/ml, respectively, and cultured for 72 hours, and then a cell viability assay kit (EX-cytox model, Daei Lab Service, Korea) solution was added according to the manufacturer's procedure, and absorbance was measured at 450nm using a microplate reader (ELISA reader) and graphed.
As a result, as shown in FIG. 1, it was confirmed that the complex extract of Stauntonia chinensis and Corni fructus was not cytotoxic.
Experimental example 1 confirmation of Effect of inhibiting expression of Prostate Specific Antigen (PSA), Androgen Receptor (AR) and 5-alpha reductase 2(5 alpha-reductase 2) proteins in a Complex extract of Japanese stauntonvine and Cornus officinalis
In order to examine whether the compound extract of Japanese stauntonvine and dogwood is effective for the treatment of prostate diseases, the expression of androgen receptor, prostate specific antigen and 5-alpha reductase 2 protein was confirmed after treating prostate cancer cells with testosterone and the compound extract of Japanese stauntonvine and dogwood.
Specifically, LNCaP cells (ATCC company, USA) were plated in 6-well plates at 5X 104The cell/well concentration was divided, and the cells were cultured at 37 ℃ for 18 hours. Then, the cultured cells were treated with testosterone (1. mu.l) and treated with the Japanese stauntonvine extract prepared in example 1 at a concentration of 25. mu.g/ml or 50. mu.g/ml, the Corni fructus extract prepared in example 2 at a concentration of 50. mu.g/ml, or the Japanese stauntonvine and Corni fructus complex extract prepared in example 3 at a concentration of 50. mu.g/ml, and then cultured for 72 hours and recovered. Instead of the extract treatment, 1 μ M testosterone was used as a positive control group (BPH), and 10 μ M Finasteride (Finasteride) as a therapeutic agent for prostatic hyperplasia was used instead of the extract, and 1 μ M testosterone was used as a control group (Finasteride). Also, sabal palm (Saw Palmetto) as a therapeutic agent for prostatic hyperplasia, 100. mu.g/ml was used as a control group (sabal palm) by treating with 1. mu.M testosterone instead of the extract. The non-treated group was used as a negative Control group (Control).
The cells recovered after the culture were treated with RIPA lysis buffer (RIPA buffer) (50mM Tris-HCl, pH 8.0, 150mM sodium chloride (sodium chloride), 1% ethylphenylpolyethylene glycol (NP-40), 0.5% sodium deoxycholate (sodium deoxycholate), 0.1% sodium dodecyl sulfate (sodium dodecyl sulfate), and a mixture of protease inhibitors (cocktail)), and the proteins were separated by centrifugation at 12000rpm for 20 minutes. Quantification was performed by the Bicinchoninic acid (BCA) method.
10g of the quantified proteins were treated with sodium dodecyl sulfate polyacrylamide (SDS-PEGE) gel to perform electrophoresis, and then the proteins were transferred to a polyvinylidene fluoride (PVDF) membrane. Thereafter, the polyvinylidene fluoride membrane was reacted in 5% skim milk (skimmilk) for about 1 hour to remove non-specifically bound proteins on the surface, and then reacted at a temperature of 4 ℃ for one day using an anti-Androgen Receptor (AR) antibody, an anti-Prostate Specific Antigen (PSA) antibody, an anti-5- α reductase 2(5 α -reductase 2) antibody, and an anti- β -actin (β -actin) antibody as primary antibodies. After that, after 1 hour of treatment at room temperature with the secondary antibody, the expression of the androgen receptor, prostate-specific antigen and antibody, and 5- α reductase 2 protein were confirmed and graphed using an ECL kit (Amersham, uk).
As a result, as shown in FIG. 2, it was confirmed that the inhibitory effect on the protein expression of androgen receptor and prostate specific antigen was significantly superior in the cell group treated with 25. mu.g/ml of the compound extract of Japanese stauntonvine and Corni fructus compared to the cell group treated with 25. mu.g/ml or 50. mu.g/ml of the extract of Japanese stauntonvine or Corni fructus alone. In addition, it was confirmed that the effect of inhibiting the expression of the protein of the androgen receptor and the prostate specific antigen was more excellent in the cell group treated with the compound extract of Japanese stauntonvine and dogwood in which the extract of Japanese stauntonvine and the extract of dogwood were mixed at a mixing ratio of 9: 1. Meanwhile, it was confirmed that the cell group treated with the compound extract of Japanese stauntonvine and dogwood has an excellent effect of inhibiting the expression of 5-alpha reductase 2 protein.
Examples of the preparation of the composition used in the present invention are shown below.
Preparation example 1 preparation of pharmaceutical preparation
1-1 preparation of powder
Powders were prepared by mixing 10mg of the complex extract of the present invention and 1g of lactose and filling into a gas-tight pad.
1-2 preparation of tablets
Tablets were prepared by mixing 0.1mg of the complex extract of the present invention, 100mg of corn starch, 100mg of lactose and 2mg of magnesium stearate, and tabletting according to a conventional tablet preparation method.
1-3 preparation of capsules
After mixing 0.1mg of the complex extract of the present invention, 100mg of corn starch, 100mg of lactose and 2mg of magnesium stearate, gelatin capsules were filled according to a common capsule preparation method to prepare capsules.
1-4 preparation of pills
After mixing 1mg of the complex extract of the present invention, 1.5g of lactose, 1g of glycerol, and 0.5g of xylitol, it was prepared in such a manner that 4g per pill was prepared according to a conventional method.
1-5 preparation of granules
After mixing 0.15mg of the complex extract of the present invention, 50mg of the soybean extract, 200mg of glucose and 600mg of starch, 100mg of 30% ethanol was added, and the mixture was dried at 60 ℃ to form granules, which were then filled in a hermetically sealed bag.
Preparation example 2 preparation of health food
A food containing the complex extract of the present invention as an active ingredient was prepared by the following method.
2-1 preparation of flour food
The compound extract of the present invention is added to flour in an amount of 0.5 to 5.0 parts by weight, and the mixture is used to prepare bread, cake, biscuit, cracker and pasta.
2-2 preparation of soups and gravies
The compound extract of the present invention is added in an amount of 0.1 to 5.0 parts by weight to a soup or a meat juice to prepare a meat-processed product, a pasta soup or a meat juice for health promotion.
2-3 preparation of ground beef
The ground beef for health promotion is prepared by adding 10 parts by weight of the composite extract of the present invention to the ground beef.
2-4 preparation of Dairy products
The composite extract of the present invention is added to milk in an amount of 5 to 10 parts by weight, and the milk is used to prepare various dairy products such as butter and ice cream.
2-5 preparation of Zen food
The product obtained by converting brown rice, barley, glutinous rice, and pearl barley into alpha-form and drying the alpha-form is baked by a known method, and then pulverized by a pulverizer to obtain a powder having a particle size of 60 mesh.
The product obtained by steaming black bean, black sesame and common perilla and drying by a known method is baked and then prepared into powder with the granularity of 60 meshes by a pulverizer.
The composite extract of the present invention was concentrated under reduced pressure in a vacuum concentrator, dried using a spray or hot air dryer to obtain a dried product, and the dried product was pulverized into a particle size of 60 mesh using a pulverizer to obtain a dried powder.
The grains, the seeds and the compound extract are prepared by mixing the following components in proportion: cereals (30 parts by weight of brown rice, 15 parts by weight of coix seed and 20 parts by weight of barley), seeds (7 parts by weight of common perilla seed, 8 parts by weight of black bean and 7 parts by weight of black sesame), composite extract (3 parts by weight) of the invention, lucid ganoderma (0.5 part by weight) and rehmannia root (0.5 part by weight).
Preparation example 3 preparation of health drink
3-1 preparation of health beverage
The extract is prepared by uniformly blending a sub-material such as liquid fructose (0.5%), oligosaccharide (2%), sucrose (2%), salt (0.5%) and water (75%) with 0.5g of the composite extract of the present invention, subjecting the mixture to flash sterilization, and packaging the mixture in a small packaging container such as a glass bottle or a polyethylene terephthalate (PET) bottle.
Preparation method of 3-2 vegetable juice
Vegetable juice was prepared by adding 0.5g of the composite extract of the present invention to 1000ml of tomato juice or carrot juice.
3-3 preparation of fruit juice
Fruit juice was prepared by adding 0.1g of the complex extract of the present invention to 1000ml of apple juice or grape juice.
Industrial applicability
The present invention relates to a composition for preventing or treating prostate diseases comprising extracts of chaenomeles japonicas and cornus officinalis as active ingredients, and more particularly, to a composition for preventing or treating prostate diseases, which comprises a compound extract of chaenomeles japonicas and cornus officinalis as an active ingredient, has no cytotoxicity, and has an excellent inhibitory effect on the expression of androgen receptors, prostate-specific antigens and prostate-specific enzymes, and thus can be effectively used as an active ingredient of a composition for preventing or treating prostate diseases.
Claims (14)
1. A pharmaceutical composition for preventing or treating prostate diseases, characterized by comprising Japanese stauntonvine extract and dogwood extract as active ingredients.
2. The pharmaceutical composition for preventing or treating prostate disorders according to claim 1, wherein said extract is selected from the group consisting of water, C1To C2Or a mixed solvent thereof.
3. The pharmaceutical composition for preventing or treating prostate diseases according to claim 2, wherein said extract is extracted with one or more solvents selected from the group consisting of water, methanol, ethanol, and a mixed solvent thereof.
4. The pharmaceutical composition for preventing or treating prostate diseases according to claim 1, wherein said extract is an extract of at least one selected from the group consisting of flowers, branches, stems, leaves, fruits, aerial parts, rhizomes, and roots.
5. The pharmaceutical composition for preventing or treating prostate diseases according to claim 1, wherein said composition comprises the extract of chaenomeles speciosa and the extract of cornus officinalis in a weight ratio of 5: 5 to 9.5: 0.5.
6. The pharmaceutical composition for preventing or treating prostate diseases according to claim 1, wherein said composition comprises the extract of chaenomeles speciosa and the extract of cornus officinalis in a weight ratio of 6: 4 to 9.5: 0.5.
7. The pharmaceutical composition according to claim 1, wherein the prostate disease is selected from the group consisting of prostate cancer, prostatic hyperplasia and prostatitis.
8. A health food for preventing or improving prostate diseases, characterized by comprising Japanese stauntonvine extract and dogwood extract as active ingredients.
9. A food composition for preventing or improving prostate diseases, characterized by comprising Japanese stauntonvine extract and dogwood extract as active ingredients.
10. A method for preventing or ameliorating a prostate disease, comprising the step of administering a pharmaceutically effective amount of Japanese stauntonvine extract and Corni fructus extract to an individual.
11. A method for treating a prostate disease, comprising the step of administering a pharmaceutically effective amount of Japanese stauntonvine extract and Corni fructus extract to an individual.
12. Use of a Japanese stauntonvine extract and a dogwood extract is characterized in that the Japanese stauntonvine extract and the dogwood extract are used as a pharmaceutical composition for preventing or treating prostate diseases.
13. Use of a Japanese stauntonvine extract and a dogwood extract is characterized in that the Japanese stauntonvine extract and the dogwood extract are used as health food for preventing or improving prostate diseases.
14. Use of a Japanese stauntonvine extract and a dogwood extract is characterized in that the Japanese stauntonvine extract and the dogwood extract are used as a food composition for preventing or improving prostate diseases.
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PCT/KR2019/009518 WO2020045838A1 (en) | 2018-08-29 | 2019-07-31 | Composition for prevention or treatment of prostate disease, containing extract from stauntonia hexaphylla and cornus officinalis as active ingredients |
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CN102302717A (en) * | 2011-09-30 | 2012-01-04 | 姜新田 | Traditional Chinese medicine composition and medical pad for treating prostatic disease, and preparation method of medical pad |
CN104173681A (en) * | 2014-09-15 | 2014-12-03 | 程惠华 | Traditional Chinese herbal medicine composition for treating castration-resistant prostate cancer |
KR101652073B1 (en) * | 2015-10-12 | 2016-08-29 | 충남대학교산학협력단 | Composition for Prevention or Treatment of Prostatic Diseases Comprising Stauntonia Hexaphylla Extract |
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KR101523586B1 (en) * | 2013-07-11 | 2015-05-29 | 상지대학교산학협력단 | Pharmaceutical composition for preventing or treating prostatic hyperplasia and preparation method thereof |
KR101675930B1 (en) * | 2015-04-02 | 2016-11-14 | 유의순 | Anticancer composition comprising herbal extracts |
KR101733477B1 (en) | 2015-06-04 | 2017-05-11 | 충남대학교산학협력단 | Composition for Prevention or Treatment of Prostatic Diseases Comprising Albizziae Cortex Extract |
KR20170117339A (en) * | 2016-04-12 | 2017-10-23 | 주식회사 케미메디 | Medical composition for preventing or treating chronic bacterial prostatitis |
KR101895548B1 (en) * | 2017-01-16 | 2018-09-06 | 대구한의대학교산학협력단 | Pharmaceutical composition comprising Korean medicinal herbs for preventing hair loss, melanizing of hair, enhancing hair growth, treating benign prostate hyperplasia and manufacturing method thereof |
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CN102302717A (en) * | 2011-09-30 | 2012-01-04 | 姜新田 | Traditional Chinese medicine composition and medical pad for treating prostatic disease, and preparation method of medical pad |
CN104173681A (en) * | 2014-09-15 | 2014-12-03 | 程惠华 | Traditional Chinese herbal medicine composition for treating castration-resistant prostate cancer |
KR101652073B1 (en) * | 2015-10-12 | 2016-08-29 | 충남대학교산학협력단 | Composition for Prevention or Treatment of Prostatic Diseases Comprising Stauntonia Hexaphylla Extract |
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